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Sample records for polarised primary cilia

  1. Secreted frizzled-related protein disrupts PCP in eye lens fiber cells that have polarised primary cilia.

    PubMed

    Sugiyama, Yuki; Stump, Richard J W; Nguyen, Anke; Wen, Li; Chen, Yongjuan; Wang, Yanshu; Murdoch, Jennifer N; Lovicu, Frank J; McAvoy, John W

    2010-02-15

    Planar cell polarity (PCP) signaling polarises cells along tissue axes. Although pathways involved are becoming better understood, outstanding issues include; (i) existence/identity of cues that orchestrate global polarisation in tissues, and (ii) the generality of the link between polarisation of primary cilia and asymmetric localisation of PCP proteins. Mammalian lenses are mainly comprised of epithelial-derived fiber cells. Concentrically arranged fibers are precisely aligned as they elongate along the anterior-posterior axis and orientate towards lens poles where they meet fibers from other segments to form characteristic sutures. We show that lens exhibits PCP, with each fiber cell having an apically situated cilium and in most cases this is polarised towards the anterior pole. Frizzled and other PCP proteins are also asymmetrically localised along the equatorial-anterior axis. Mutations in core PCP genes Van Gogh-like 2 and Celsr1 perturb oriented fiber alignment and suture formation. Suppression of the PCP pathway by overexpressing Sfrp2 shows that whilst local groups of fibers are often similarly oriented, they lack global orientation; consequently when local groups of fibers with different orientations meet they form multiple, small, ectopic suture-like configurations. This indicates that this extracellular inhibitor disrupts a global polarising signal that utilises a PCP-mediated mechanism to coordinate the global alignment and orientation of fibers to lens poles. PMID:19968984

  2. Mechanical Properties of Primary Cilia

    NASA Astrophysics Data System (ADS)

    Battle, Christopher; Schmidt, Christoph F.

    2013-03-01

    Recent studies have shown that the primary cilium, long thought to be a vestigial cellular appendage with no function, is involved in a multitude of sensory functions. One example, interesting from both a biophysical and medical standpoint, is the primary cilium of kidney epithelial cells, which acts as a mechanosensitive flow sensor. Genetic defects in ciliary function can cause, e.g., polycystic kidney disease (PKD). The material properties of these non-motile, microtubule-based 9 +0 cilia, and the way they are anchored to the cell cytoskeleton, are important to know if one wants to understand the mechano-electrochemical response of these cells, which is mediated by their cilia. We have probed the mechanical properties, boundary conditions, and dynamics of the cilia of MDCK cells using optical traps and DIC/fluorescence microscopy. We found evidence for both elastic relaxation of the cilia themselves after bending and for compliance in the intracellular anchoring structures. Angular and positional fluctuations of the cilia reflect both thermal excitations and cellular driving forces.

  3. Primary cilia and forebrain development.

    PubMed

    Willaredt, Marc August; Tasouri, Evangelia; Tucker, Kerry L

    2013-01-01

    With a microtubule-based axoneme supporting its plasma membrane-ensheathed projection from the basal body of almost all cell types in the human body, and present in only one copy per cell, the primary cilium can be considered an organelle sui generis. Although it was first observed and recorded in histological studies from the late 19th century, the tiny structure was essentially forgotten for many decades. In the past ten years, however, scientists have turned their eyes once again upon primary cilia and realized that they are very important for the development of almost all organs in the mammalian body, especially those dependent upon the signaling from members Hedgehog family, such as Indian and Sonic hedgehog. In this review, we outline the roles that primary cilia play in forebrain development, not just in the crucial transduction of Sonic hedgehog signaling, but also new results showing that cilia are important for cell cycle progression in proliferating neural precursors. We will focus upon cerebral cortex development but will also discuss the importance of cilia for the embryonic hippocampus, olfactory bulb, and diencephalon. PMID:23085524

  4. Immunofluorescent staining of septins in primary cilia.

    PubMed

    Kim, M S; Froese, C D; Xie, H; Trimble, W S

    2016-01-01

    Primary cilia are cellular antennae that receive and transduce extracellular cues. These microtubule-rich structures are comprised of at least three distinct ciliary compartments: basal bodies, transition zone, and axoneme. Septins have been implicated in cilia function at the transition zone, but accumulating evidence suggests that they localize predominantly within the axoneme. Here, we describe three fixation conditions that preserve the substructure of primary cilia and demonstrate known ciliary proteins that localize to these distinct ciliary substructures. Finally, we show immunostaining and live microscopy methods to detect septins within the axoneme. PMID:27473914

  5. Primary Cilia in Pancreatic Development and Disease

    PubMed Central

    Lodh, Sukanya; O’Hare, Elizabeth A.; Zaghloul, Norann A.

    2014-01-01

    Primary cilia and their anchoring basal bodies are important regulators of a growing list of signaling pathways. Consequently, dysfunction in proteins associated with these structures results in perturbation of the development and function of a spectrum of tissue and cell types. Here, we review the role of cilia in mediating the development and function of the pancreas. We focus on ciliary regulation of major pathways involved in pancreatic development, including Shh, Wnt, TGF-β, Notch, and fibroblast growth factor. We also discuss pancreatic phenotypes associated with ciliary dysfunction, including pancreatic cysts and defects in glucose homeostasis, and explore the potential role of cilia in such defects. PMID:24864023

  6. Primary cilia utilize glycoprotein-dependent adhesion mechanisms to stabilize long-lasting cilia-cilia contacts

    PubMed Central

    2012-01-01

    Background The central tenet of cilia function is sensing and transmitting information. The capacity to directly contact extracellular surfaces would empower primary cilia to probe the environment for information about the nature and location of nearby surfaces. It has been well established that flagella and other motile cilia perform diverse cellular functions through adhesion. We hypothesized that mammalian primary cilia also interact with the extracellular environment through direct physical contact. Methods We identified cilia in rod photoreceptors and cholangiocytes in fixed mouse tissues and examined the structures that these cilia contact in vivo. We then utilized an MDCK cell culture model to characterize the nature of the contacts we observed. Results In retina and liver tissue, we observed that cilia from nearby cells touch one another. Using MDCK cells, we found compelling evidence that these contacts are stable adhesions that form bridges between two cells, or networks between many cells. We examined the nature and duration of the cilia-cilia contacts and discovered primary cilia movements that facilitate cilia-cilia encounters. Stable adhesions form as the area of contact expands from a single point to a stretch of tightly bound, adjacent cilia membranes. The cilia-cilia contacts persisted for hours and were resistant to several harsh treatments such as proteases and DTT. Unlike many other cell adhesion mechanisms, calcium was not required for the formation or maintenance of cilia adhesion. However, swainsonine, which blocks maturation of N-linked glycoproteins, reduced contact formation. We propose that cellular control of adhesion maintenance is active because cilia adhesion did not prevent cell division; rather, contacts dissolved during mitosis as cilia were resorbed. Conclusions The demonstration that mammalian primary cilia formed prolonged, direct, physical contacts supports a novel paradigm: that mammalian primary cilia detect features of the

  7. Detection of primary cilia in human glioblastoma

    PubMed Central

    Sarkisian, Matthew R.; Siebzehnrubl, Dorit; Hoang-Minh, Lan; Deleyrolle, Loic; Silver, Daniel J.; Siebzehnrubl, Florian A.; Guadiana, Sarah M.; Srivinasan, Gayathri; Semple-Rowland, Susan; Harrison, Jeffrey K.; Steindler, Dennis A.; Reynolds, Brent A.

    2015-01-01

    Glioblastoma (GBM) is the most common malignant adult brain tumor and carries a poor prognosis due to primary and acquired resistance. While many cellular features of GBM have been documented, it is unclear if cells within these tumors extend a primary cilium, an organelle whose associated signaling pathways may regulate proliferation, migration, and survival of neural precursor and tumor cells. Using immunohistochemical and electron microscopy (EM) techniques, we screened human GBM tumor biopsies and primary cell lines for cilia. Immunocytochemical staining of five primary GBM cell lines revealed that between 8 and 25 % of the cells in each line possessed gamma tubulin-positive basal bodies from which extended acetylated, alpha-tubulin-positive axonemes. EM analyses confirmed the presence of cilia at the cell surface and revealed that their axonemes contained organized networks of microtubules, a structural feature consistent with our detection of IFT88 and Arl13b, two trafficked cilia proteins, along the lengths of the axonemes. Notably, cilia were detected in each of 23 tumor biopsies (22 primary and 1 recurrent) examined. These cilia were distributed across the tumor landscape including regions proximal to the vasculature and within necrotic areas. Moreover, ciliated cells within these tumors co-stained with Ki67, a marker for actively dividing cells, and ZEB1, a transcription factor that is upregulated in GBM and linked to tumor initiation, invasion, and chemoresistance. Collectively, our data show that subpopulations of cells within human GBM tumors are ciliated. In view of mounting evidence supporting roles of primary cilia in tumor initiation and propagation, it is likely that further study of the effects of cilia on GBM tumor cell function will improve our understanding of GBM pathogenesis and may provide new directions for GBM treatment strategies. PMID:24510433

  8. The Roles of Primary cilia in Polycystic Kidney Disease

    PubMed Central

    Kathem, Sarmed H.; Mohieldin, Ashraf M.; Nauli, Surya M.

    2014-01-01

    Autosomal dominant polycystic kidney disease (ADPKD) is an inherited genetic disorder that results in progressive renal cyst formation with ultimate loss of renal function and other systemic disorders. These systemic disorders include abnormalities in cardiovascular, portal, pancreatic and gastrointestinal systems. ADPKD is considered to be among the ciliopathy diseases due to the association with abnormal primary cilia function. In order to understand the full course of primary cilia and its association with ADPKD, the structure, functions and role of primary cilia have been meticulously investigated. As a result, the focus on primary cilia has emerged to support the vital roles of primary cilia in ADPKD. The primary cilia have been shown to have not only a mechanosensory function but also a chemosensory function. Both structural and functional defects in primary cilia result in cystic kidney disease and vascular hypertension. Thus, the mechanosenory and chemosensory functions will be analyzed in regards to ADPKD. PMID:25599087

  9. Autophagy and primary cilia: dual interplay.

    PubMed

    Pampliega, Olatz; Cuervo, Ana Maria

    2016-04-01

    Primary cilia are microtubule-based organelles for sensing of the extracellular milieu and transducing this information into the cell through a variety of molecular signaling pathways. Functioning of the primary cilium has been recently connected to autophagy, a pathway for degradation of cellular components in lysosomes. Autophagy regulates the length of the cilia by removing proteins required for ciliogenesis, a phenomenon that is molecularly different if performed by basal autophagy or when autophagy is induced in response to various stressors. Here we review the current knowledge about the dual interaction between autophagy and ciliogenesis, and discuss the potential role that deregulated ciliary autophagy could have in pathologies with alterations in autophagy and ciliogenesis. PMID:26826446

  10. The emerging face of primary cilia

    PubMed Central

    Zaghloul, Norann A.; Brugmann, Samantha A.

    2011-01-01

    Primary cilia are microtubule-based organelles that serve as hubs for the transduction of various developmental signaling pathways including Hedgehog, Wnt, FGF and PDGF. Ciliary dysfunction contributes to a range of disorders, collectively known as the ciliopathies. Recently, interest has grown in these syndromes, particularly among craniofacial biologists, as many known and putative ciliopathies have severe craniofacial defects. Herein we discuss the current understanding of ciliary biology and craniofacial development in an attempt to gain insight into the molecular etiology for craniofacial ciliopathies, and uncover a characteristic ciliopathic craniofacial gestalt. PMID:21305689

  11. Adenylate cyclase regulates elongation of mammalian primary cilia

    SciTech Connect

    Ou, Young; Ruan, Yibing; Cheng, Min; Moser, Joanna J.; Rattner, Jerome B.; Hoorn, Frans A. van der

    2009-10-01

    The primary cilium is a non-motile microtubule-based structure that shares many similarities with the structures of flagella and motile cilia. It is well known that the length of flagella is under stringent control, but it is not known whether this is true for primary cilia. In this study, we found that the length of primary cilia in fibroblast-like synoviocytes, either in log phase culture or in quiescent state, was confined within a range. However, when lithium was added to the culture to a final concentration of 100 mM, primary cilia of synoviocytes grew beyond this range, elongating to a length that was on average approximately 3 times the length of untreated cilia. Lithium is a drug approved for treating bipolar disorder. We dissected the molecular targets of this drug, and observed that inhibition of adenylate cyclase III (ACIII) by specific inhibitors mimicked the effects of lithium on primary cilium elongation. Inhibition of GSK-3{beta} by four different inhibitors did not induce primary cilia elongation. ACIII was found in primary cilia of a variety of cell types, and lithium treatment of these cell types led to their cilium elongation. Further, we demonstrate that different cell types displayed distinct sensitivities to the lithium treatment. However, in all cases examined primary cilia elongated as a result of lithium treatment. In particular, two neuronal cell types, rat PC-12 adrenal medulla cells and human astrocytes, developed long primary cilia when lithium was used at or close to the therapeutic relevant concentration (1-2 mM). These results suggest that the length of primary cilia is controlled, at least in part, by the ACIII-cAMP signaling pathway.

  12. ADENYLATE CYCLASE REGULATES ELONGATION OF MAMMALIAN PRIMARY CILIA

    PubMed Central

    Ou, Young; Ruan, Yibing; Cheng, Min; Moser, Joanna J.; Rattner, Jerome B.; van der Hoorn, Frans A.

    2011-01-01

    The primary cilium is a non-motile microtubule-based structure that shares many similarities with the structures of flagella and motile cilia. It is well known that the length of flagella is under stringent control, but it is not known whether this is true for primary cilia. In this study, we found that the length of primary cilia in fibroblast-like synoviocytes, either in log phase culture or in quiescent state, was confined within a range. However, when lithium was added to the culture to a final concentration of 100 mM, primary cilia of synoviocytes grew beyond this range, elongating to a length that was on average approximately 3 times the length of untreated cilia. Lithium is a drug approved for treating bipolar disorder. We dissected the molecular targets of this drug, and observed that inhibition of adenylate cyclase III (ACIII) by specific inhibitors mimicked the effects of lithium on primary cilium elongation. Inhibition of GSK-3β by four different inhibitors did not induce primary cilia elongation. ACIII was found in primary cilia of a variety of cell types, and lithium treatment of these cell types led to their cilium elongation. Further, we demonstrate that different cell types displayed distinct sensitivities to the lithium treatment. However, in all cases examined primary cilia elongated as a result of lithium treatment. In particular, two neuronal cell types, rat PC-12 adrenal medulla cells and human astrocytes, developed long primary cilia when lithium was used at or close to the therapeutic relevant concentration (1–2 mM). These results suggest that the length of primary cilia is controlled, at least in part, by the ACIII-cAMP signaling pathway. PMID:19576885

  13. CLEM Methods for Studying Primary Cilia.

    PubMed

    Macaluso, Frank P; Perumal, Geoffrey S; Kolstrup, Johan; Satir, Peter

    2016-01-01

    CLEM (correlated light and electron microscope) imaging is a highly useful technique for examining primary cilia. With CLEM, it is possible to determine the distribution of tagged proteins along the ciliary membrane and axoneme with high precision. Scanning electron microscopy (SEM) permits measurement of ciliary length and orientation in relation to nearby cellular structures in a 3D image; in optimal cases, this can be combined with superresolution microscopy of selected ciliary components as they enter or leave the cilium. This chapter discusses CLEM methods. In the method described in detail, samples are completely processed for sequential fluorescence and SEM observation. This method is ideal for robust antibody localization and minimizes image manipulation in correlating the fluorescent and SEM images. Alternative methods prepare samples for fluorescence imaging followed by processing for SEM then observation in the SEM. This method is ideal for optimal fluorescence imaging, particularly live cell imaging. PMID:27514923

  14. Electrical Signaling in Motile and Primary Cilia

    PubMed Central

    Kleene, Steven J.; Van Houten, Judith L.

    2014-01-01

    Cilia are highly conserved for their structure and also for their sensory functions. They serve as antennae for extracellular information. Whether the cilia are motile or not, they respond to environmental mechanical and chemical stimuli and send signals to the cell body. The information from extracellular stimuli is commonly converted to electrical signals through the repertoire of ion-conducting channels in the ciliary membrane, which results in changes in concentrations of ions, especially calcium ions, in the cilia. These changes, in turn, affect motility and the ability of the signaling pathways in the cilia and cell body to carry on the signal transduction. We review here the activities of ion channels in cilia in animals from protists to vertebrates. PMID:25892740

  15. Visualization of live primary cilia dynamics using fluorescence microscopy

    PubMed Central

    Ott, Carolyn; Lippincott-Schwartz, Jennifer

    2013-01-01

    Here we describe methods that are useful for exploring the formation and functions of primary cilia in living cells. First we describe multiple protocols for visualizing solitary cilia that extend away from the cell body. Primary cilia collect, synthesize, and transmit information about the extracellular space into the cell body to promote critical cellular responses. Problems with cilia formation or function can lead to dramatic changes in cell physiology. These methods can be used to assess cilia formation and length, the location of the cilium relative to other cellular structures, and localization of specific proteins to the cilium. The second protocol describes how to quantify movement of fluorescent molecules within the cilium. The microtubules that form the structural scaffold of the cilium are also critical avenues for kinesin and dynein-mediated movement of proteins within the cilium. Assessing intraflagellar dynamics can provide insight into mechanisms of ciliary-mediated signal perception and transmission. PMID:23208547

  16. Primary cilia are not calcium-responsive mechanosensors.

    PubMed

    Delling, M; Indzhykulian, A A; Liu, X; Li, Y; Xie, T; Corey, D P; Clapham, D E

    2016-03-31

    Primary cilia are solitary, generally non-motile, hair-like protrusions that extend from the surface of cells between cell divisions. Their antenna-like structure leads naturally to the assumption that they sense the surrounding environment, the most common hypothesis being sensation of mechanical force through calcium-permeable ion channels within the cilium. This Ca(2+)-responsive mechanosensor hypothesis for primary cilia has been invoked to explain a large range of biological responses, from control of left-right axis determination in embryonic development to adult progression of polycystic kidney disease and some cancers. Here we report the complete lack of mechanically induced calcium increases in primary cilia, in tissues upon which this hypothesis has been based. We developed a transgenic mouse, Arl13b-mCherry-GECO1.2, expressing a ratiometric genetically encoded calcium indicator in all primary cilia. We then measured responses to flow in primary cilia of cultured kidney epithelial cells, kidney thick ascending tubules, crown cells of the embryonic node, kinocilia of inner ear hair cells, and several cell lines. Cilia-specific Ca(2+) influxes were not observed in physiological or even highly supraphysiological levels of fluid flow. We conclude that mechanosensation, if it originates in primary cilia, is not via calcium signalling. PMID:27007841

  17. Primary cilia and autophagic dysfunction in Huntington's disease.

    PubMed

    Kaliszewski, M; Knott, A B; Bossy-Wetzel, E

    2015-09-01

    Huntington's disease (HD) is an inherited, neurodegenerative disorder caused by a single-gene mutation: a CAG expansion in the huntingtin (HTT) gene that results in production of a mutated protein, mutant HTT, with a polyglutamine tail (polyQ-HTT). Although the molecular pathways of polyQ-HTT toxicity are not fully understood, because protein misfolding and aggregation are central features of HD, it has long been suspected that cellular housekeeping processes such as autophagy might be important to disease pathology. Indeed, multiple lines of research have identified abnormal autophagy in HD, characterized generally by increased autophagic induction and inefficient clearance of substrates. To date, the origin of autophagic dysfunction in HD remains unclear and the search for actors involved continues. To that end, recent studies have suggested a bidirectional relationship between autophagy and primary cilia, signaling organelles of most mammalian cells. Interestingly, primary cilia structure is defective in HD, suggesting a potential link between autophagic dysfunction, primary cilia and HD pathogenesis. In addition, because polyQ-HTT also accumulates in primary cilia, the possibility exists that primary cilia might play additional roles in HD: perhaps by disrupting signaling pathways or acting as a reservoir for secretion and propagation of toxic, misfolded polyQ-HTT fragments. Here, we review recent research suggesting potential links between autophagy, primary cilia and HD and speculate on possible pathogenic mechanisms and future directions for the field. PMID:26160070

  18. Analysis of primary cilia in the developing mouse brain.

    PubMed

    Paridaen, Judith T M L; Huttner, Wieland B; Wilsch-Bräuninger, Michaela

    2015-01-01

    Stem and progenitor cells in the developing mammalian brain are highly polarized cells that carry a primary cilium protruding into the brain ventricles. Here, cilia detect signals present in the cerebrospinal fluid that fills the ventricles. Recently, striking observations have been made regarding the dynamics of primary cilia in mitosis and cilium reformation after cell division. In neural progenitors, primary cilia are not completely disassembled during cell division, and some ciliary membrane remnant can be inherited by one daughter cell that tends to maintain a progenitor fate. Furthermore, newborn differentiating cells grow a primary cilium on their basolateral plasma membrane, in spite of them possessing apical membrane and adherens junctions, and thus change the environment to which the primary cilium is exposed. These phenomena are proposed to be involved in cell fate determination and delamination of daughter cells in conjunction with the production of neurons. Here, we describe several methods that can be used to study the structure, localization, and dynamics of primary cilia in the developing mouse brain; these include time-lapse imaging of live mouse embryonic brain tissues, and analysis of primary cilia structure and localization using correlative light- and electron- and serial-block-face scanning electron microscopy. PMID:25837388

  19. Ins and outs of GPCR signaling in primary cilia.

    PubMed

    Schou, Kenneth Bødtker; Pedersen, Lotte Bang; Christensen, Søren Tvorup

    2015-09-01

    Primary cilia are specialized microtubule-based signaling organelles that convey extracellular signals into a cellular response in most vertebrate cell types. The physiological significance of primary cilia is underscored by the fact that defects in assembly or function of these organelles lead to a range of severe diseases and developmental disorders. In most cell types of the human body, signaling by primary cilia involves different G protein-coupled receptors (GPCRs), which transmit specific signals to the cell through G proteins to regulate diverse cellular and physiological events. Here, we provide an overview of GPCR signaling in primary cilia, with main focus on the rhodopsin-like (class A) and the smoothened/frizzled (class F) GPCRs. We describe how such receptors dynamically traffic into and out of the ciliary compartment and how they interact with other classes of ciliary GPCRs, such as class B receptors, to control ciliary function and various physiological and behavioral processes. Finally, we discuss future avenues for developing GPCR-targeted drug strategies for the treatment of ciliopathies. PMID:26297609

  20. Primary cilia and Gli3 activity regulate cerebral cortical size

    PubMed Central

    Wilson, Sandra L.; Wilson, John P.; Wang, Chengbing; Wang, Baolin; McConnell, Susan K.

    2012-01-01

    During neural development, patterning, neurogenesis and overall growth are highly regulated and coordinated between different brain regions. Here, we show that primary cilia and the regulation of Gli activity, are necessary for the normal expansion of the cerebral cortex. We show that loss of Kif3a, an important functional component of primary cilia, leads to the degeneration of primary cilia, marked overgrowth of the cortex, and altered cell cycle kinetics within cortical progenitors. The G1 phase of the cell cycle is shortened through a mechanism likely involving reduced Gli3 activity and a resulting increase in expression of cyclin D1 and Fgf15. The defects in Gli3 activity alone are sufficient to accelerate cell cycle kinetics and cause the molecular changes seen in brains that lack cilia. Finally, we show that levels of full-length and repressor Gli3 proteins are tightly regulated during normal development and correlate with changes in expression of two known Shh-target genes, CyclinD1 and Fgf15, and with the normal lengthening of the cell cycle during corticogenesis. These data suggest that Gli3 activity is regulated through the primary cilium to control cell cycle length in the cortex and thus determine cortical size. PMID:21976438

  1. Intracellular and extracellular forces drive primary cilia movement.

    PubMed

    Battle, Christopher; Ott, Carolyn M; Burnette, Dylan T; Lippincott-Schwartz, Jennifer; Schmidt, Christoph F

    2015-02-01

    Primary cilia are ubiquitous, microtubule-based organelles that play diverse roles in sensory transduction in many eukaryotic cells. They interrogate the cellular environment through chemosensing, osmosensing, and mechanosensing using receptors and ion channels in the ciliary membrane. Little is known about the mechanical and structural properties of the cilium and how these properties contribute to ciliary perception. We probed the mechanical responses of primary cilia from kidney epithelial cells [Madin-Darby canine kidney-II (MDCK-II)], which sense fluid flow in renal ducts. We found that, on manipulation with an optical trap, cilia deflect by bending along their length and pivoting around an effective hinge located below the basal body. The calculated bending rigidity indicates weak microtubule doublet coupling. Primary cilia of MDCK cells lack interdoublet dynein motors. Nevertheless, we found that the organelles display active motility. 3D tracking showed correlated fluctuations of the cilium and basal body. These angular movements seemed random but were dependent on ATP and cytoplasmic myosin-II in the cell cortex. We conclude that force generation by the actin cytoskeleton surrounding the basal body results in active ciliary movement. We speculate that actin-driven ciliary movement might tune and calibrate ciliary sensory functions. PMID:25605896

  2. Intracellular and extracellular forces drive primary cilia movement

    PubMed Central

    Battle, Christopher; Ott, Carolyn M.; Burnette, Dylan T.; Lippincott-Schwartz, Jennifer; Schmidt, Christoph F.

    2015-01-01

    Primary cilia are ubiquitous, microtubule-based organelles that play diverse roles in sensory transduction in many eukaryotic cells. They interrogate the cellular environment through chemosensing, osmosensing, and mechanosensing using receptors and ion channels in the ciliary membrane. Little is known about the mechanical and structural properties of the cilium and how these properties contribute to ciliary perception. We probed the mechanical responses of primary cilia from kidney epithelial cells [Madin–Darby canine kidney-II (MDCK-II)], which sense fluid flow in renal ducts. We found that, on manipulation with an optical trap, cilia deflect by bending along their length and pivoting around an effective hinge located below the basal body. The calculated bending rigidity indicates weak microtubule doublet coupling. Primary cilia of MDCK cells lack interdoublet dynein motors. Nevertheless, we found that the organelles display active motility. 3D tracking showed correlated fluctuations of the cilium and basal body. These angular movements seemed random but were dependent on ATP and cytoplasmic myosin-II in the cell cortex. We conclude that force generation by the actin cytoskeleton surrounding the basal body results in active ciliary movement. We speculate that actin-driven ciliary movement might tune and calibrate ciliary sensory functions. PMID:25605896

  3. Primary cilia and kidney injury: current research status and future perspectives

    PubMed Central

    Wang, Shixuan

    2013-01-01

    Cilia, membrane-enclosed organelles protruding from the apical side of cells, can be divided into two classes: motile and primary cilia. During the past decades, motile cilia have been intensively studied. However, it was not until the 1990s that people began to realize the importance of primary cilia as cellular-specific sensors, particularly in kidney tubular epithelial cells. Furthermore, accumulating evidence indicates that primary cilia may be involved in the regulation of cell proliferation, differentiation, apoptosis, and planar cell polarity. Many signaling pathways, such as Wnt, Notch, Hedgehog, and mammalian target of rapamycin, have been located to the primary cilia. Thus primary cilia have been regarded as a hub that integrates signals from the extracellular environment. More importantly, dysfunction of this organelle may contribute to the pathogenesis of a large spectrum of human genetic diseases, named ciliopathies. The significance of primary cilia in acquired human diseases such as hypertension and diabetes has gradually drawn attention. Interestingly, recent reports disclosed that cilia length varies during kidney injury, and shortening of cilia enhances the sensitivity of epithelial cells to injury cues. This review briefly summarizes the current status of cilia research and explores the potential mechanisms of cilia-length changes during kidney injury as well as provides some thoughts to allure more insightful ideas and promotes the further study of primary cilia in the context of kidney injury. PMID:23904226

  4. Primary Cilia Regulate Branching Morphogenesis During Mammary Gland Development

    PubMed Central

    McDermott, Kimberly M.; Liu, Bob Y.; Tlsty, Thea D.; Pazour, Gregory J.

    2010-01-01

    Summary During mammary gland development an epithelial bud undergoes branching morphogenesis to expand into a continuous tree-like network of branched ducts [1]. The process involves multiple cell types that are coordinated by hormones and growth factors coupled with signaling events including Wnt and Hedgehog [2-5]. Primary cilia play key roles in the development of many organs by coordinating extracellular signaling (Wnt, Hedgehog) with cellular physiology [6-8]. During mammary development, we find cilia on luminal epithelial, myoepithelial and stromal cells during early branching morphogenesis when epithelial ducts extend into the fat pad and undergo branching morphogenesis. When branching is complete, cilia disappear from luminal epithelial cells but remain on myoepithelial and stromal cells. Ciliary dysfunction caused by intraflagellar transport (IFT) defects results in branching defects. These include decreased ductal extension and decreased secondary and tertiary branching along with reduced lobular-alveolar development during pregnancy and lactation. We find increased canonical Wnt and decreased Hedgehog signaling in the mutant glands, which is consistent with the role of cilia in regulating these pathways [6-11]. In mammary gland and other organs, increased canonical Wnt [12-14] and decreased Hedgehog [15, 16] signaling decreases branching morphogenesis suggesting that Wnt and Hedgehog signaling connect ciliary dysfunction to branching defects. PMID:20381354

  5. Characterization of cancer stem cells and primary cilia in medulloblastoma.

    PubMed

    Gate, David; Danielpour, Moise; Bannykh, Serguei; Town, Terrence

    2015-01-01

    Medulloblastoma, a tumor of the cerebellum, is the most common pediatric central nervous system malignancy. These tumors are etiologically linked to mutations in the Sonic hedgehog (Shh) pathway, which signals through the primary, non-motile cilium. The growth of these aggressive tumors relies on self-renewal of tumor-propagating cells known as cancer stem cells (CSCs). Previous reports have implicated CD133-expressing cells as CSCs in brain tumors, while those expressing CD15 have been shown to propagate medulloblastoma. Here, we demonstrate that CD133+ and CD15+ cells are distinct medulloblastoma populations. CD15+ cells comprise approximately 0.5-1% of total human medulloblastoma cells, display CSC properties in culture and are detected in the Smoothened A1 transgenic mouse model of medulloblastoma. Additionally, we report on a medulloblastoma patient with enriched CD15+ cells in recurrent vs primary medulloblastoma. We also demonstrate that human medulloblastoma cells critically rely on establishment of primary cilia to drive Shh-mediated cell division. Primary cilia are found in external granule cells of human fetal cerebellum and in 12/14 medulloblastoma samples. Yet, CD15+ medulloblastoma cells lack primary cilia, suggesting that this CSC population signals independently of Shh. These results are important when considering the effects of current and prospective treatment modalities on medulloblastoma CSC populations. PMID:25921740

  6. Oscillatory Fluid Flow Influences Primary Cilia and Microtubule Mechanics

    PubMed Central

    Espinha, Lina C.; Hoey, David A.; Fernandes, Paulo R.; Rodrigues, Hélder C.; Jacobs, Christopher R.

    2014-01-01

    Many tissues are sensitive to mechanical stimuli; however, the mechanotransduction mechanism used by cells remains unknown in many cases. The primary cilium is a solitary, immotile microtubule-based extension present on nearly every mammalian cell which extends from the basal body. The cilium is a mechanosensitive organelle and has been shown to transduce fluid flow-induced shear stress in tissues such as the kidney and bone. The majority of microtubules assemble from the mother centriole (basal body), contributing significantly to the anchoring of the primary cilium. Several studies have attempted to quantify the number of microtubules emanating from the basal body and the results vary depending on the cell type. It has also been shown that cellular response to shear stress depends on microtubular integrity. This study hypothesizes that changing the microtubule attachment of primary cilia in response to a mechanical stimulus could change primary cilia mechanics and, possibly, mechanosensitivity. Oscillatory fluid flow was applied to two different cell types and the microtubule attachment to the ciliary base was quantified. For the first time, an increase in microtubules around primary cilia both with time and shear rate in response to oscillatory fluid flow stimulation was demonstrated. Moreover, it is presented that the primary cilium is required for this loading-induced cellular response. This study has demonstrated a new role for the cilium in regulating alterations in the cytoplasmic microtubule network in response to mechanical stimulation, and therefore provides a new insight into how cilia may regulate its mechanics and thus the cells mechanosensitivity. PMID:25044764

  7. Primary cilia in energy balance signaling and metabolic disorder

    PubMed Central

    Lee, Hankyu; Song, Jieun; Jung, Joo Hyun; Ko, Hyuk Wan

    2015-01-01

    Energy homeostasis in our body system is maintained by balancing the intake and expenditure of energy. Excessive accumulation of fat by disrupting the balance system causes overweight and obesity, which are increasingly becoming global health concerns. Understanding the pathogenesis of obesity focused on studying the genes related to familial types of obesity. Recently, a rare human genetic disorder, ciliopathy, links the role for genes regulating structure and function of a cellular organelle, the primary cilium, to metabolic disorder, obesity and type II diabetes. Primary cilia are microtubule based hair-like membranous structures, lacking motility and functions such as sensing the environmental cues, and transducing extracellular signals within the cells. Interestingly, the subclass of ciliopathies, such as Bardet-Biedle and Alström syndrome, manifest obesity and type II diabetes in human and mouse model systems. Moreover, studies on genetic mouse model system indicate that more ciliary genes affect energy homeostasis through multiple regulatory steps such as central and peripheral actions of leptin and insulin. In this review, we discuss the latest findings in primary cilia and metabolic disorders, and propose the possible interaction between primary cilia and the leptin and insulin signal pathways which might enhance our understanding of the unambiguous link of a cell’s antenna to obesity and type II diabetes. [BMB Reports 2015; 48(12): 647-654] PMID:26538252

  8. Function and regulation of primary cilia and intraflagellar transport proteins in the skeleton

    PubMed Central

    Yuan, Xue; Serra, Rosa A.; Yang, Shuying

    2014-01-01

    Primary cilia are microtubule-based organelles that project from the cell surface to enable transduction of various developmental signaling pathways. The process of intraflagellar transport (IFT) is crucial for the building and maintenance of primary cilia. Ciliary dysfunction has been found in a range of disorders called ciliopathies, some of which display severe skeletal dysplasias. In recent years, interest has grown in uncovering the function of primary cilia/IFT proteins in bone development, mechanotransduction, and cellular regulation. We summarize recent advances in understanding the function of cilia and IFT proteins in the regulation of cell differentiation in osteoblasts, osteocytes, chondrocytes, and mesenchymal stem cells (MSCs). We also discuss the mechanosensory function of cilia and IFT proteins in bone cells, cilia orientation, and other functions of cilia in chondrocytes. PMID:24961486

  9. Molecular complexes that direct rhodopsin transport to primary cilia

    PubMed Central

    Wang, Jing; Deretic, Dusanka

    2013-01-01

    Rhodopsin is a key molecular constituent of photoreceptor cells, yet understanding of how it regulates photoreceptor membrane trafficking and biogenesis of light-sensing organelles, the rod outer segments (ROS) is only beginning to emerge. Recently identified sequence of well-orchestrated molecular interactions of rhodopsin with the functional networks of Arf and Rab GTPases at multiple stages of intracellular targeting fits well into the complex framework of the biogenesis and maintenance of primary cilia, of which the ROS is one example. This review will discuss the latest progress in dissecting the molecular complexes that coordinate rhodopsin incorporation into ciliary-targeted carriers with the recruitment and activation of membrane tethering complexes and regulators of fusion with the periciliary plasma membrane. In addition to revealing the fundamental principals of ciliary membrane renewal, recent advances also provide molecular insight into the ways by which disruptions of the exquisitely orchestrated interactions lead to cilia dysfunction and result in human retinal dystrophies and syndromic diseases that affect multiple organs, including the eyes. PMID:24135424

  10. Unilateral nephrectomy elongates primary cilia in the remaining kidney via reactive oxygen species

    PubMed Central

    Han, Sang Jun; Jang, Hee-Seong; Kim, Jee In; Lipschutz, Joshua H.; Park, Kwon Moo

    2016-01-01

    The length of primary cilia is associated with normal cell and organ function. In the kidney, the change of functional cilia length/mass is associated with various diseases such as ischemia/reperfusion injury, polycystic kidney disease, and congenital solitary kidney. Here, we investigate whether renal mass reduction affects primary cilia length and function. To induce renal mass reduction, mice were subjected to unilateral nephrectomy (UNx). UNx increased kidney weight and superoxide formation in the remaining kidney. Primary cilia were elongated in proximal tubule cells, collecting duct cells and parietal cells of the remaining kidney. Mn(III) Tetrakis (1-methyl-4-pyridyl) porphyrin (MnTMPyP), an antioxidant, reduced superoxide formation in UNx-mice and prevented the elongation of primary cilia. UNx increased the expression of phosphorylated ERK, p21, and exocyst complex members Sec8 and Sec10, in the remaining kidney, and these increases were prevented by MnTMPyP. In MDCK, a kidney tubular epithelial cell line, cells, low concentrations of H2O2 treatment elongated primary cilia. This H2O2-induced elongation of primary cilia was also prevented by MnTMPyP treatment. Taken together, these data demonstrate that kidney compensation, induced by a reduction of renal mass, results in primary cilia elongation, and this elongation is associated with an increased production of reactive oxygen species (ROS). PMID:26923764

  11. Primary cilia are highly oriented with respect to collagen direction and long axis of extensor tendon.

    PubMed

    Donnelly, Eve; Ascenzi, Maria-Grazia; Farnum, Cornelia

    2010-01-01

    Skeletal tissues adapt to their mechanical environments by modulating gene expression, cell metabolism, and extracellular matrix (ECM) architecture; however, the mechanosensory mechanisms for these processes are incompletely understood. Primary cilia have emerged as critical components of the cellular mechanosensory apparatus and have been hypothesized to participate in establishment of cellular and ECM orientation, but their function in skeletal tissues is just beginning to be examined. Here we focused on tendon, a tissue with an oriented matrix that is ideal for analysis of spatial relationships between primary cilia and the ECM. The objective of this study was to characterize the incidence and orientation of tenocyte primary cilia in their native ECM. Primary cilia, nuclei, and collagen were analyzed three-dimensionally in immunofluorescently labeled rat extensor tendon using multiphoton microscopy and semiautomated morphometry. Primary cilia were observed in 64% of tenocytes. The cilia were highly oriented with respect to the ECM: cilia were aligned parallel to the collagen fibers and the long axis of the tendon. This study represents the first quantification of the in situ incidence and orientation of primary cilia in tendon. PMID:19603516

  12. Developmental changes in primary cilia in the mouse tooth germ and oral cavity.

    PubMed

    Hisamoto, Meri; Goto, Marie; Muto, Mami; Nio-Kobayashi, Junko; Iwanaga, Toshihiko; Yokoyama, Atsuro

    2016-01-01

    The primary cilium, a sensory apparatus, functions as both a chemical and mechanical sensor to receive environmental stimuli. The present study focused on the primary cilia in the epithelialmesenchymal interaction during tooth development. We examined the localization and direction of projection of primary cilia in the tooth germ and oral cavity of mice by immunohistochemical observation. Adenylyl cyclase 3 (ACIII)-immunolabeled cilia were visible in the inner/outer enamel epithelium of molars at the fetal stage and then conspicuously developed in the odontoblast layer postnatally. The primary cilia in ameloblasts and odontoblasts-shown by the double staining of acetylated tubulin and γ-tubulin-were regularly arranged from postnatal Day12, projecting apart from each other. The periodontal ligament possessed ACIII-positive cilia, which gathered on both sides of the dentin/cement and alveolar bone in postnatal days. In the oral cavity, numerous long primary cilia immunoreactive for ACIII were condensed at subepithelial stromal cells in the oral processes in fetuses, while postnatally a small number of short cilia were dispersed throughout the stroma of the oral cavity. These findings suggest that the primary cilia showing stage- and regionspecific morphology are involved in the epithelial-mesenchymal interaction during tooth development via mechano- and/or chemoreception for growth factors. PMID:27356608

  13. Surface topography regulates wnt signaling through control of primary cilia structure in mesenchymal stem cells

    NASA Astrophysics Data System (ADS)

    McMurray, R. J.; Wann, A. K. T.; Thompson, C. L.; Connelly, J. T.; Knight, M. M.

    2013-12-01

    The primary cilium regulates cellular signalling including influencing wnt sensitivity by sequestering β-catenin within the ciliary compartment. Topographic regulation of intracellular actin-myosin tension can control stem cell fate of which wnt is an important mediator. We hypothesized that topography influences mesenchymal stem cell (MSC) wnt signaling through the regulation of primary cilia structure and function. MSCs cultured on grooves expressed elongated primary cilia, through reduced actin organization. siRNA inhibition of anterograde intraflagellar transport (IFT88) reduced cilia length and increased active nuclear β-catenin. Conversely, increased primary cilia assembly in MSCs cultured on the grooves was associated with decreased levels of nuclear active β-catenin, axin-2 induction and proliferation, in response to wnt3a. This negative regulation, on grooved topography, was reversed by siRNA to IFT88. This indicates that subtle regulation of IFT and associated cilia structure, tunes the wnt response controlling stem cell differentiation.

  14. Comparative study of the primary cilia in thyrocytes of adult mammals.

    PubMed

    Utrilla, J C; Gordillo-Martínez, F; Gómez-Pascual, A; Fernández-Santos, J M; Garnacho, C; Vázquez-Román, V; Morillo-Bernal, J; García-Marín, R; Jiménez-García, A; Martín-Lacave, I

    2015-10-01

    Since their discovery in different human tissues by Zimmermann in 1898, primary cilia have been found in the vast majority of cell types in vertebrates. Primary cilia are considered to be cellular antennae that occupy an ideal cellular location for the interpretation of information both from the environment and from other cells. To date, in mammalian thyroid gland, primary cilia have been found in the thyrocytes of humans and dogs (fetuses and adults) and in rat embryos. The present study investigated whether the existence of this organelle in follicular cells is a general event in the postnatal thyroid gland of different mammals, using both immunolabeling by immunofluorescence and electron microscopy. Furthermore, we aimed to analyse the presence of primary cilia in various thyroid cell lines. According to our results, primary cilia are present in the adult thyroid gland of most mammal species we studied (human, pig, guinea pig and rabbit), usually as a single copy per follicular cell. Strikingly, they were not found in rat or mouse thyroid tissues. Similarly, cilia were also observed in all human thyroid cell lines tested, both normal and neoplastic follicular cells, but not in cultured thyrocytes of rat origin. We hypothesize that primary cilia could be involved in the regulation of normal thyroid function through specific signaling pathways. Nevertheless, further studies are needed to shed light on the permanence of these organelles in the thyroid gland of most species during postnatal life. PMID:26228270

  15. Direct recording and molecular identification of the calcium channel of primary cilia

    NASA Astrophysics Data System (ADS)

    Decaen, Paul G.; Delling, Markus; Vien, Thuy N.; Clapham, David E.

    2013-12-01

    A primary cilium is a solitary, slender, non-motile protuberance of structured microtubules (9+0) enclosed by plasma membrane. Housing components of the cell division apparatus between cell divisions, primary cilia also serve as specialized compartments for calcium signalling and hedgehog signalling pathways. Specialized sensory cilia such as retinal photoreceptors and olfactory cilia use diverse ion channels. An ion current has been measured from primary cilia of kidney cells, but the responsible genes have not been identified. The polycystin proteins (PC and PKD), identified in linkage studies of polycystic kidney disease, are candidate channels divided into two structural classes: 11-transmembrane proteins (PKD1, PKD1L1 and PKD1L2) remarkable for a large extracellular amino terminus of putative cell adhesion domains and a G-protein-coupled receptor proteolytic site, and the 6-transmembrane channel proteins (PKD2, PKD2L1 and PKD2L2; TRPPs). Evidence indicates that the PKD1 proteins associate with the PKD2 proteins via coiled-coil domains. Here we use a transgenic mouse in which only cilia express a fluorophore and use it to record directly from primary cilia, and demonstrate that PKD1L1 and PKD2L1 form ion channels at high densities in several cell types. In conjunction with an accompanying manuscript, we show that the PKD1L1-PKD2L1 heteromeric channel establishes the cilia as a unique calcium compartment within cells that modulates established hedgehog pathways.

  16. Tubulin glycylases are required for primary cilia, control of cell proliferation and tumor development in colon

    PubMed Central

    Rocha, Cecilia; Papon, Laura; Cacheux, Wulfran; Marques Sousa, Patricia; Lascano, Valeria; Tort, Olivia; Giordano, Tiziana; Vacher, Sophie; Lemmers, Benedicte; Mariani, Pascale; Meseure, Didier; Medema, Jan Paul; Bièche, Ivan; Hahne, Michael; Janke, Carsten

    2014-01-01

    TTLL3 and TTLL8 are tubulin glycine ligases catalyzing posttranslational glycylation of microtubules. We show here for the first time that these enzymes are required for robust formation of primary cilia. We further discover the existence of primary cilia in colon and demonstrate that TTLL3 is the only glycylase in this organ. As a consequence, colon epithelium shows a reduced number of primary cilia accompanied by an increased rate of cell division in TTLL3-knockout mice. Strikingly, higher proliferation is compensated by faster tissue turnover in normal colon. In a mouse model for tumorigenesis, lack of TTLL3 strongly promotes tumor development. We further demonstrate that decreased levels of TTLL3 expression are linked to the development of human colorectal carcinomas. Thus, we have uncovered a novel role for tubulin glycylation in primary cilia maintenance, which controls cell proliferation of colon epithelial cells and plays an essential role in colon cancer development. PMID:25180231

  17. Tubulin glycylases are required for primary cilia, control of cell proliferation and tumor development in colon.

    PubMed

    Rocha, Cecilia; Papon, Laura; Cacheux, Wulfran; Marques Sousa, Patricia; Lascano, Valeria; Tort, Olivia; Giordano, Tiziana; Vacher, Sophie; Lemmers, Benedicte; Mariani, Pascale; Meseure, Didier; Medema, Jan Paul; Bièche, Ivan; Hahne, Michael; Janke, Carsten

    2014-10-01

    TTLL3 and TTLL8 are tubulin glycine ligases catalyzing posttranslational glycylation of microtubules. We show here for the first time that these enzymes are required for robust formation of primary cilia. We further discover the existence of primary cilia in colon and demonstrate that TTLL3 is the only glycylase in this organ. As a consequence, colon epithelium shows a reduced number of primary cilia accompanied by an increased rate of cell division in TTLL3-knockout mice. Strikingly, higher proliferation is compensated by faster tissue turnover in normal colon. In a mouse model for tumorigenesis, lack of TTLL3 strongly promotes tumor development. We further demonstrate that decreased levels of TTLL3 expression are linked to the development of human colorectal carcinomas. Thus, we have uncovered a novel role for tubulin glycylation in primary cilia maintenance, which controls cell proliferation of colon epithelial cells and plays an essential role in colon cancer development. PMID:25180231

  18. Primary Cilia Integrate Hedgehog and Wnt Signaling during Tooth Development

    PubMed Central

    Liu, B.; Chen, S.; Cheng, D.; Jing, W.; Helms, J.A.

    2014-01-01

    Many ciliopathies have clinical features that include tooth malformations but how these defects come about is not clear. Here we show that genetic deletion of the motor protein Kif3a in dental mesenchyme results in an arrest in odontogenesis. Incisors are completely missing, and molars are enlarged in Wnt1Cre+Kif3afl/fl embryos. Although amelogenesis and dentinogenesis initiate in the molar tooth bud, both processes terminate prematurely. We demonstrate that loss of Kif3a in dental mesenchyme results in loss of Hedgehog signaling and gain of Wnt signaling in this same tissue. The defective dental mesenchyme then aberrantly signals to the dental epithelia, which prompts an up-regulation in the Hedgehog and Wnt responses in the epithelia and leads to multiple attempts at invagination and an expanded enamel organ. Thus, the primary cilium integrates Hedgehog and Wnt signaling between dental epithelia and mesenchyme, and this cilia-dependent integration is required for proper tooth development. PMID:24659776

  19. Primary Cilia on Horizontal Basal Cells Regulate Regeneration of the Olfactory Epithelium

    PubMed Central

    Joiner, Ariell M.; Green, Warren W.; McIntyre, Jeremy C.; Allen, Benjamin L.; Schwob, James E.

    2015-01-01

    The olfactory epithelium (OE) is one of the few tissues to undergo constitutive neurogenesis throughout the mammalian lifespan. It is composed of multiple cell types including olfactory sensory neurons (OSNs) that are readily replaced by two populations of basal stem cells, frequently dividing globose basal cells and quiescent horizontal basal cells (HBCs). However, the precise mechanisms by which these cells mediate OE regeneration are unclear. Here, we show for the first time that the HBC subpopulation of basal stem cells uniquely possesses primary cilia that are aligned in an apical orientation in direct apposition to sustentacular cell end feet. The positioning of these cilia suggests that they function in the detection of growth signals and/or differentiation cues. To test this idea, we generated an inducible, cell type-specific Ift88 knock-out mouse line (K5rtTA;tetOCre;Ift88fl/fl) to disrupt cilia formation and maintenance specifically in HBCs. Surprisingly, the loss of HBC cilia did not affect the maintenance of the adult OE but dramatically impaired the regeneration of OSNs following lesion. Furthermore, the loss of cilia during development resulted in a region-specific decrease in neurogenesis, implicating HBCs in the establishment of the OE. Together, these results suggest a novel role for primary cilia in HBC activation, proliferation, and differentiation. SIGNIFICANCE STATEMENT We show for the first time the presence of primary cilia on a quiescent population of basal stem cells, the horizontal basal cells (HBCs), in the olfactory epithelium (OE). Importantly, our data demonstrate that cilia on HBCs are necessary for regeneration of the OE following injury. Moreover, the disruption of HBC cilia alters neurogenesis during the development of the OE, providing evidence that HBCs participate in the establishment of this tissue. These data suggest that the mechanisms of penetrance for ciliopathies in the OE extend beyond that of defects in olfactory sensory

  20. Modulation of primary cilia length by melanin-concentrating hormone receptor 1.

    PubMed

    Hamamoto, Akie; Yamato, Shogo; Katoh, Yohei; Nakayama, Kazuhisa; Yoshimura, Kentaro; Takeda, Sen; Kobayashi, Yuki; Saito, Yumiko

    2016-06-01

    Melanin-concentrating hormone (MCH) receptor 1 (MCHR1) is a class A G-protein-coupled receptor (GPCR). The MCH-MCHR1 system has been implicated in the regulation of feeding, emotional processing, and sleep in rodents. Recent work revealed that MCHR1 is selectively expressed in neuronal primary cilia of the central nervous system. Cilia have various chemosensory functions in many types of cell, and ciliary dysfunction is associated with ciliopathies such as polycystic kidney disease and obesity. Although dynamic modulation of neuronal cilia length is observed in obese mice, the functional interaction of neuronal ciliary GPCR and its endogenous ligand has not yet been elucidated. We report here that MCH treatment significantly reduced cilia length in hTERT-RPE1 cells (hRPE1 cells) transfected with MCHR1. Quantitative analyses indicated that MCH-induced cilia shortening progressed in a dose-dependent manner with an EC50 lower than 1nM when cells were treated for 6h. Although the assembly and disassembly of primary cilia are tightly coupled to the cell cycle, cell cycle reentry was not a determinant of MCH-induced cilia shortening. We confirmed that MCH elicited receptor internalization, Ca(2+) mobilization, ERK and Akt phosphorylation, and inhibition of cyclic AMP accumulation in MCHR1-expressing hRPE1 cells. Among these diverse pathways, we revealed that Gi/o-dependent Akt phosphorylation was an important component in the initial stage of MCH-induced cilia length shortening. Furthermore, induction of fewer cilia by Kif3A siRNA treatment significantly decreased the MCH-mediated phosphorylation of Akt, indicating the functional importance of the MCHR1-Akt pathway in primary cilia. Taken together, the present data suggest that the MCH-MCHR1 axis may modulate the sensitivity of cells to external environments by controlling the cilia length. Therefore, further characterization of MCHR1 as a ciliary GPCR will provide a potential molecular mechanism to link cilia length

  1. Lithium treatment elongates primary cilia in the mouse brain and in cultured cells

    SciTech Connect

    Miyoshi, Ko; Kasahara, Kyosuke; Miyazaki, Ikuko; Asanuma, Masato

    2009-10-30

    The molecular mechanisms underlying the therapeutic effects of lithium, a first-line antimanic mood stabilizer, have not yet been fully elucidated. Treatment of the algae Chlamydomonas reinhardtii with lithium has been shown to induce elongation of their flagella, which are analogous structures to vertebrate cilia. In the mouse brain, adenylyl cyclase 3 (AC3) and certain neuropeptide receptors colocalize to the primary cilium of neuronal cells, suggesting a chemosensory function for the primary cilium in the nervous system. Here we show that lithium treatment elongates primary cilia in the mouse brain and in cultured cells. Brain sections from mice chronically fed with Li{sub 2}CO{sub 3} were subjected to immunofluorescence study. Primary cilia carrying both AC3 and the receptor for melanin-concentrating hormone (MCH) were elongated in the dorsal striatum and nucleus accumbens of lithium-fed mice, as compared to those of control animals. Moreover, lithium-treated NIH3T3 cells and cultured striatal neurons exhibited elongation of the primary cilia. The present results provide initial evidence that a psychotropic agent can affect ciliary length in the central nervous system, and furthermore suggest that lithium exerts its therapeutic effects via the upregulation of cilia-mediated MCH sensing. These findings thus contribute novel insights into the pathophysiology of bipolar mood disorder and other psychiatric diseases.

  2. Hippocampal and Cortical Primary Cilia Are Required for Aversive Memory in Mice

    PubMed Central

    Yazdi, S. M. Zaki R.; McNair, Andrew D.; Kippe, Jordyn M.; Croyle, Mandy J.; Kraft, Timothy W.; Yoder, Bradley K.

    2014-01-01

    It has been known for decades that neurons throughout the brain possess solitary, immotile, microtubule based appendages called primary cilia. Only recently have studies tried to address the functions of these cilia and our current understanding remains poor. To determine if neuronal cilia have a role in behavior we specifically disrupted ciliogenesis in the cortex and hippocampus of mice through conditional deletion of the Intraflagellar Transport 88 (Ift88) gene. The effects on learning and memory were analyzed using both Morris Water Maze and fear conditioning paradigms. In comparison to wild type controls, cilia mutants displayed deficits in aversive learning and memory and novel object recognition. Furthermore, hippocampal neurons from mutants displayed an altered paired-pulse response, suggesting that loss of IFT88 can alter synaptic properties. A variety of other behavioral tests showed no significant differences between conditional cilia mutants and controls. This type of conditional allele approach could be used to distinguish which behavioral features of ciliopathies arise due to defects in neural development and which result from altered cell physiology. Ultimately, this could lead to an improved understanding of the basis for the cognitive deficits associated with human cilia disorders such as Bardet-Biedl syndrome, and possibly more common ailments including depression and schizophrenia. PMID:25184295

  3. RC/BTB2 is Essential for Formation of Primary Cilia in Mammalian Cells

    PubMed Central

    Zhang, Ling; Li, Wei; Ni, Jin; Wu, Jinghua; Liu, Junping; Zhang, Zhengang; Zhang, Yong; Li, Hongfei; Shi, Yuqin; Teves, Maria E; Song, Shizheng; Strauss, Jerome F.; Zhang, Zhibing

    2016-01-01

    RC/BTB2 is a binding partner of sperm associated antigen 16S (SPAG16S), which is regulator of spermiogenesis in mice, a process during which sperm flagella are formed. The expression of Rc/btb2 is also regulated by multicilin, a protein that controls ciliogenesis. Given that mouse Rc/btb2 mRNA is not only expressed in tissues bearing motile cilia, but also in tissues without motile cilia, we investigated whether RC/BTB2 plays a role in the general process of ciliogenesis by studying two somatic cells lines that have primary cilia, NIH3T3 and IMCD3. We discovered that the subcellular localization of RT/BTB2 in the NIH3T3 and IMCD3 cells encompasses the pathway for ciliogenesis. RC/BTB2 was found in the Golgi bodies and centrosomes, two key structures essential for normal ciliogenesis. Knockdown of Rc/btb2 gene expression in these cell lines disrupted ciliogenesis. The percentage of cells with primary cilia was significantly reduced in stable cell lines transduced with specific Rc/btb2 shRNA viruses compared to the control cells. When cilia were formed in the knockdown cells, they were significantly shorter than those in the control cells. Knockdown of Rc/btb2 expression did not affect cell proliferation and the cell cycle. Exogenous expression of RC/BTB2 in these stable knockdown cells restored ciliogenesis. These findings suggest that RC/BTB2 is a necessary component of the process of formation of primary cilia in somatic cells, perhaps through the transportation of cargos from Golgi bodies to centrosomes for cilia assembling. PMID:25762510

  4. An Essential Role for Dermal Primary Cilia in Hair Follicle Morphogenesis

    PubMed Central

    Lehman, Jonathan; Laag, Essam; Michaud, Edward J.; Yoder, Bradley K.

    2009-01-01

    The primary cilium is a microtubule-based organelle implicated as an essential component of a number of signaling pathways. It is present on cells throughout the mammalian body; however, its functions in most tissues remain largely unknown. Herein we demonstrate that primary cilia are present on cells in murine skin and hair follicles throughout morphogenesis and during hair follicle cycling in postnatal life. Using the Cre-lox system, we disrupted cilia assembly in the ventral dermis and evaluated the effects on hair follicle development. Mice with disrupted dermal cilia have severe hypotrichosis (lack of hair) in affected areas. Histological analyses reveal that most follicles in the mutants arrest at stage 2 of hair development and have small or absent dermal condensates. This phenotype is reminiscent of that seen in the skin of mice lacking Shh or Gli2. In situ hybridization and quantitative RT-PCR analysis indicates that the hedgehog pathway is downregulated in the dermis of the cilia mutant hair follicles. Thus, these data establish cilia as a critical signaling component required for normal hair morphogenesis and suggest that this organelle is needed on cells in the dermis for reception of signals such as sonic hedgehog. PMID:18987668

  5. Loss of primary cilia upregulates renal hypertrophic signaling and promotes cystogenesis.

    PubMed

    Bell, P Darwin; Fitzgibbon, Wayne; Sas, Kelli; Stenbit, Antine E; Amria, May; Houston, Amber; Reichert, Ryan; Gilley, Sandra; Siegal, Gene P; Bissler, John; Bilgen, Mehmet; Chou, Peter Cheng-te; Guay-Woodford, Lisa; Yoder, Brad; Haycraft, Courtney J; Siroky, Brian

    2011-05-01

    Primary cilia dysfunction alters renal tubular cell proliferation and differentiation and associates with accelerated cyst formation in polycystic kidney disease. However, the mechanism leading from primary ciliary dysfunction to renal cyst formation is unknown. We hypothesize that primary cilia prevent renal cyst formation by suppressing pathologic tubular cell hypertrophy and proliferation. Unilateral nephrectomy initiates tubular cell hypertrophy and proliferation in the contralateral kidney and provides a tool to examine primary cilia regulation of renal hypertrophy. Conditional knockout of the primary cilia ift88 gene leads to delayed, adult-onset renal cystic disease, which provides a window of opportunity to conduct unilateral nephrectomy and examine downstream kinetics of renal hypertrophy and cyst formation. In wild-type animals, unilateral nephrectomy activated the mTOR pathway and produced appropriate structural and functional hypertrophy without renal cyst formation. However, in ift88 conditional knockout animals, unilateral nephrectomy triggered increased renal hypertrophy and accelerated renal cyst formation, leading to renal dysfunction. mTOR signaling also increased compared with wild-type animals, suggesting a mechanistic cascade starting with primary ciliary dysfunction, leading to excessive mTOR signaling and renal hypertrophic signaling and culminating in cyst formation. These data suggest that events initiating hypertrophic signaling, such as structural or functional loss of renal mass, may accelerate progression of adult polycystic kidney disease toward end-stage renal disease. PMID:21493775

  6. Primary cilia expression in bone marrow in response to mechanical stimulation in explant bioreactor culture.

    PubMed

    Coughlin, T R; Schiavi, J; Alyssa Varsanik, M; Voisin, M; Birmingham, E; Haugh, M G; McNamara, L M; Niebur, G L

    2016-01-01

    Bone marrow contains a multitude of mechanically sensitive cells that may participate in mechanotransduction. Primary cilia are sensory organelles expressed on mesenchymal stem cells (MSCs), osteoblasts, osteocytes, and other cell types that sense fluid flow in monolayer culture. In marrow, cilia could similarly facilitate the sensation of relative motion between adjacent cells or interstitial fluid. The goal of this study was to determine the response of cilia to mechanical stimulation of the marrow. Bioreactors were used to supply trabecular bone explants with low magnitude mechanical stimulation (LMMS) of 0.3 ×g at 30 Hz for 1 h/d, 5 d/week, inducing shear stresses in the marrow. Four groups were studied: unstimulated (UNSTIM), stimulated (LMMS), and with and without chloral hydrate (UNSTIM+CH and LMMS+CH, respectively), which was used to disrupt cilia. After 19 days of culture, immunohistochemistry for acetylated α-tubulin revealed that more cells expressed cilia in culture compared to in vivo controls. Stimulation decreased the number of cells expressing cilia in untreated explants, but not in CH-treated explants. MSCs represented a greater fraction of marrow cells in the untreated explants than CH-treated explants. MSCs harvested from the stimulated groups were more proliferative than in the unstimulated explants, but this effect was absent from CH treated explants. In contrast to the marrow, neither LMMS nor CH treatment affected bone formation as measured by mineralising surface. Computational models indicated that LMMS does not induce bone strain, and the reported effects were thus attributed to shear stress in the marrow. From a clinical perspective, genetic or pharmaceutical alterations of cilia expression may affect marrow health and function. PMID:27434268

  7. Hedgehog signaling regulates myelination in the peripheral nervous system through primary cilia.

    PubMed

    Yoshimura, Kentaro; Takeda, Sen

    2012-02-01

    Myelination is an essential prerequisite for the nervous system to transmit an impulse efficiently by a saltatory conduction. In the peripheral nervous system (PNS), Schwann cells (SCs) engage in myelination. However, a detailed molecular mechanism underlying myelination still remains unclear. In this study, we hypothesized that the primary cilia of SCs are the regulators of Hedgehog (Hh) signaling-mediated myelination. To confirm our hypothesis, we used mouse dorsal root ganglion (DRG)/SC co-cultures, wherein the behavior of SCs could be analyzed by maintaining the interaction of SCs with DRG neurons. Under these conditions, SCs had primary cilia, and Hh signaling molecules accumulated on the primary cilia. When the SCs were stimulated by the addition of desert hedgehog or smoothened agonist, formation of myelin segments on the DRG axons was facilitated. On the contrary, upon administration of cyclopamine, an inhibitor of Hh signaling, myelin segments became comparable to those of controls. Of note, the ratio of SCs harboring primary cilium reached the highest point during the early phase of myelination. Furthermore, the strongest effects of Hh on myelination were encountered during the same stage. These results collectively indicate that Hh signaling regulates myelin formation through primary cilia in the PNS. PMID:22101064

  8. CCDC65 Mutation Causes Primary Ciliary Dyskinesia with Normal Ultrastructure and Hyperkinetic Cilia

    PubMed Central

    Horani, Amjad; Brody, Steven L.; Ferkol, Thomas W.; Shoseyov, David; Wasserman, Mollie G.; Ta-shma, Asaf; Wilson, Kate S.; Bayly, Philip V.; Amirav, Israel; Cohen-Cymberknoh, Malena; Dutcher, Susan K.; Elpeleg, Orly; Kerem, Eitan

    2013-01-01

    Background Primary ciliary dyskinesia (PCD) is a genetic disorder characterized by impaired ciliary function, leading to chronic sinopulmonary disease. The genetic causes of PCD are still evolving, while the diagnosis is often dependent on finding a ciliary ultrastructural abnormality and immotile cilia. Here we report a novel gene associated with PCD but without ciliary ultrastructural abnormalities evident by transmission electron microscopy, but with dyskinetic cilia beating. Methods Genetic linkage analysis was performed in a family with a PCD subject. Gene expression was studied in Chlamydomonas reinhardtii and human airway epithelial cells, using RNA assays and immunostaining. The phenotypic effects of candidate gene mutations were determined in primary culture human tracheobronchial epithelial cells transduced with gene targeted shRNA sequences. Video-microscopy was used to evaluate cilia motion. Results A single novel mutation in CCDC65, which created a termination codon at position 293, was identified in a subject with typical clinical features of PCD. CCDC65, an orthologue of the Chlamydomonas nexin-dynein regulatory complex protein DRC2, was localized to the cilia of normal nasal epithelial cells but was absent in those from the proband. CCDC65 expression was up-regulated during ciliogenesis in cultured airway epithelial cells, as was DRC2 in C. reinhardtii following deflagellation. Nasal epithelial cells from the affected individual and CCDC65-specific shRNA transduced normal airway epithelial cells had stiff and dyskinetic cilia beating patterns compared to control cells. Moreover, Gas8, a nexin-dynein regulatory complex component previously identified to associate with CCDC65, was absent in airway cells from the PCD subject and CCDC65-silenced cells. Conclusion Mutation in CCDC65, a nexin-dynein regulatory complex member, resulted in a frameshift mutation and PCD. The affected individual had altered cilia beating patterns, and no detectable

  9. Asymmetric Distribution of Primary Cilia Allocates Satellite Cells for Self-Renewal.

    PubMed

    Jaafar Marican, Nur Hayati; Cruz-Migoni, Sara B; Borycki, Anne-Gaëlle

    2016-06-14

    Regeneration of vertebrate skeletal muscles requires satellite cells, a population of stem cells that are quiescent in normal conditions and divide, differentiate, and self-renew upon activation triggered by exercise, injury, and degenerative diseases. Satellite cell self-renewal is essential for long-term tissue homeostasis, and previous work has identified a number of external cues that control this process. However, little is known of the possible intrinsic control mechanisms of satellite cell self-renewal. Here, we show that quiescent satellite cells harbor a primary cilium, which is rapidly disassembled upon entry into the cell cycle. Contrasting with a commonly accepted belief, cilia reassembly does not occur uniformly in cells exiting the cell cycle. We found that primary cilia reassemble preferentially in cells committed to self-renew, and disruption of cilia reassembly causes a specific deficit in self-renewing satellite cells. These observations indicate that primary cilia provide an intrinsic cue essential for satellite cell self-renewal. PMID:27161363

  10. Cyst growth, polycystins, and primary cilia in autosomal dominant polycystic kidney disease.

    PubMed

    Lee, Seung Hun; Somlo, Stefan

    2014-06-01

    The primary cilium of renal epithelia acts as a transducer of extracellular stimuli. Polycystin (PC)1 is the protein encoded by the PKD1 gene that is responsible for the most common and severe form of autosomal dominant polycystic kidney disease (ADPKD). PC1 forms a complex with PC2 via their respective carboxy-terminal tails. Both proteins are expressed in the primary cilia. Mutations in either gene affect the normal architecture of renal tubules, giving rise to ADPKD. PC1 has been proposed as a receptor that modulates calcium signals via the PC2 channel protein. The effect of PC1 dosage has been described as the rate-limiting modulator of cystic disease. Reduced levels of PC1 or disruption of the balance in PC1/PC2 level can lead to the clinical features of ADPKD, without complete inactivation. Recent data show that ADPKD resulting from inactivation of polycystins can be markedly slowed if structurally intact cilia are also disrupted at the same time. Despite the fact that no single model or mechanism from these has been able to describe exclusively the pathogenesis of cystic kidney disease, these findings suggest the existence of a novel cilia-dependent, cyst-promoting pathway that is normally repressed by polycystin function. The results enable us to rethink our current understanding of genetics and cilia signaling pathways of ADPKD. PMID:26877954

  11. GPR88 Reveals a Discrete Function of Primary Cilia as Selective Insulators of GPCR Cross-Talk

    PubMed Central

    Marley, Aaron; Choy, Regina Wai-Yan; von Zastrow, Mark

    2013-01-01

    A number of G protein-coupled receptors (GPCRs) localize to primary cilia but the functional significance of cilia to GPCR signaling remains incompletely understood. We investigated this question by focusing on the D1 dopamine receptor (D1R) and beta-2 adrenergic receptor (B2AR), closely related catecholamine receptors that signal by stimulating production of the diffusible second messenger cyclic AMP (cAMP) but differ in localization relative to cilia. D1Rs robustly concentrate on cilia of IMCD3 cells, as shown previously in other ciliated cell types, but disrupting cilia did not affect D1R surface expression or ability to mediate a concentration-dependent cAMP response. By developing a FRET-based biosensor suitable for resolving intra- from extra- ciliary cAMP changes, we found that the D1R-mediated cAMP response is not restricted to cilia and extends into the extra-ciliary cytoplasm. Conversely the B2AR, which we show here is effectively excluded from cilia, also generated a cAMP response in both ciliary and extra-ciliary compartments. We identified a distinct signaling effect of primary cilia through investigating GPR88, an orphan GPCR that is co-expressed with the D1R in brain, and which we show here is targeted to cilia similarly to the D1R. In ciliated cells, mutational activation of GPR88 strongly reduced the D1R-mediated cAMP response but did not affect the B2AR-mediated response. In marked contrast, in non-ciliated cells, GPR88 was distributed throughout the plasma membrane and inhibited the B2AR response. These results identify a discrete ‘insulating’ function of primary cilia in conferring selectivity on integrated catecholamine signaling through lateral segregation of receptors, and suggest a cellular activity of GPR88 that might underlie its effects on dopamine-dependent behaviors. PMID:23936473

  12. Identification of G Protein-Coupled Receptors (GPCRs) in Primary Cilia and Their Possible Involvement in Body Weight Control.

    PubMed

    Omori, Yoshihiro; Chaya, Taro; Yoshida, Satoyo; Irie, Shoichi; Tsujii, Toshinori; Furukawa, Takahisa

    2015-01-01

    Primary cilia are sensory organelles that harbor various receptors such as G protein-coupled receptors (GPCRs). We analyzed subcellular localization of 138 non-odorant GPCRs. We transfected GPCR expression vectors into NIH3T3 cells, induced ciliogenesis by serum starvation, and observed subcellular localization of GPCRs by immunofluorescent staining. We found that several GPCRs whose ligands are involved in feeding behavior, including prolactin-releasing hormone receptor (PRLHR), neuropeptide FF receptor 1 (NPFFR1), and neuromedin U receptor 1 (NMUR1), localized to the primary cilia. In addition, we found that a short form of dopamine receptor D2 (DRD2S) is efficiently transported to the primary cilia, while a long form of dopamine receptor D2 (DRD2L) is rarely transported to the primary cilia. Using an anti-Prlhr antibody, we found that Prlhr localized to the cilia on the surface of the third ventricle in the vicinity of the hypothalamic periventricular nucleus. We generated the Npy2r-Cre transgenic mouse line in which Cre-recombinase is expressed under the control of the promoter of Npy2r encoding a ciliary GPCR. By mating Npy2r-Cre mice with Ift80 flox mice, we generated Ift80 conditional knockout (CKO) mice in which Npy2r-positive cilia were diminished in number. We found that Ift80 CKO mice exhibited a body weight increase. Our results suggest that Npy2r-positive cilia are important for body weight control. PMID:26053317

  13. Primary cilia in stem cells and neural progenitors are regulated by neutral sphingomyelinase 2 and ceramide

    PubMed Central

    He, Qian; Wang, Guanghu; Wakade, Sushama; Dasgupta, Somsankar; Dinkins, Michael; Kong, Ji Na; Spassieva, Stefka D.; Bieberich, Erhard

    2014-01-01

    We show here that human embryonic stem (ES) and induced pluripotent stem cell–derived neuroprogenitors (NPs) develop primary cilia. Ciliogenesis depends on the sphingolipid ceramide and its interaction with atypical PKC (aPKC), both of which distribute to the primary cilium and the apicolateral cell membrane in NP rosettes. Neural differentiation of human ES cells to NPs is concurrent with a threefold elevation of ceramide—in particular, saturated, long-chain C16:0 ceramide (N-palmitoyl sphingosine) and nonsaturated, very long chain C24:1 ceramide (N-nervonoyl sphingosine). Decreasing ceramide levels by inhibiting ceramide synthase or neutral sphingomyelinase 2 leads to translocation of membrane-bound aPKC to the cytosol, concurrent with its activation and the phosphorylation of its substrate Aurora kinase A (AurA). Inhibition of aPKC, AurA, or a downstream target of AurA, HDAC6, restores ciliogenesis in ceramide-depleted cells. Of importance, addition of exogenous C24:1 ceramide reestablishes membrane association of aPKC, restores primary cilia, and accelerates neural process formation. Taken together, these results suggest that ceramide prevents activation of HDAC6 by cytosolic aPKC and AurA, which promotes acetylation of tubulin in primary cilia and, potentially, neural processes. This is the first report on the critical role of ceramide generated by nSMase2 in stem cell ciliogenesis and differentiation. PMID:24694597

  14. Primary cilia mechanosensing triggers autophagy-regulated cell volume control.

    PubMed

    Takacs, Zsuzsanna; Proikas-Cezanne, Tassula

    2016-05-27

    The primary cilium and the process of autophagy are thought to be in a functionally reciprocal relationship. In further support of this link, fluid flow sensing by the primary cilium is now shown to induce autophagy, which in turn regulates the volume of kidney epithelial cells. PMID:27230529

  15. Endocytic recycling protein EHD1 regulates primary cilia morphogenesis and SHH signaling during neural tube development

    PubMed Central

    Bhattacharyya, Sohinee; Rainey, Mark A; Arya, Priyanka; Dutta, Samikshan; George, Manju; Storck, Matthew D.; McComb, Rodney D.; Muirhead, David; Todd, Gordon L.; Gould, Karen; Datta, Kaustubh; Waes, Janee Gelineau-van; Band, Vimla; Band, Hamid

    2016-01-01

    Members of the four-member C-terminal EPS15-Homology Domain-containing (EHD) protein family play crucial roles in endocytic recycling of cell surface receptors from endosomes to the plasma membrane. In this study, we show that Ehd1 gene knockout in mice on a predominantly B6 background is embryonic lethal. Ehd1-null embryos die at mid-gestation with a failure to complete key developmental processes including neural tube closure, axial turning and patterning of the neural tube. We found that Ehd1-null embryos display short and stubby cilia on the developing neuroepithelium at embryonic day 9.5 (E9.5). Loss of EHD1 also deregulates the ciliary SHH signaling with Ehd1-null embryos displaying features indicative of increased SHH signaling, including a significant downregulation in the formation of the GLI3 repressor and increase in the ventral neuronal markers specified by SHH. Using Ehd1-null MEFS we found that EHD1 protein co-localizes with the SHH receptor Smoothened in the primary cilia upon ligand stimulation. Under the same conditions, EHD1 was shown to co-traffic with Smoothened into the developing primary cilia and we identify EHD1 as a direct binding partner of Smoothened. Overall, our studies identify the endocytic recycling regulator EHD1 as a novel regulator of the primary cilium-associated trafficking of Smoothened and Hedgehog signaling. PMID:26884322

  16. Pulsed electromagnetic fields promote osteoblast mineralization and maturation needing the existence of primary cilia.

    PubMed

    Yan, Juan-Li; Zhou, Jian; Ma, Hui-Ping; Ma, Xiao-Ni; Gao, Yu-Hai; Shi, Wen-Gui; Fang, Qing-Qing; Ren, Qian; Xian, Cory J; Chen, Ke-Ming

    2015-03-15

    Although pulsed electromagnetic fields (PEMFs) have been approved as a therapy for osteoporosis, action mechanisms and optimal parameters are elusive. To determine the optimal intensity, exposure effects of 50 Hz PEMFs of 0.6-3.6 mT (0.6 interval at 90 min/day) were investigated on proliferation and osteogenic differentiation of cultured calvarial osteoblasts. All intensity groups stimulated proliferation significantly with the highest effect at 0.6 mT. The 0.6 mT group also obtained the optimal osteogenic effect as demonstrated by the highest ALP activity, ALP(+) CFU-f colony formation, nodule mineralization, and expression of COL-1 and BMP-2. To verify our hypothesis that the primary cilia are the cellular sensors for PEMFs, osteoblasts were also transfected with IFT88 siRNA or scrambled control, and osteogenesis-promoting effects of 0.6 mT PEMFs were found abrogated when primary cilia were inhibited by IFT88 siRNA. Thus primary cilia of osteoblasts play an indispensable role in mediating PEMF osteogenic effect in vitro. PMID:25661534

  17. Active stochastic stress fluctuations in the cell cytoskeleton stir the cell and activate primary cilia

    NASA Astrophysics Data System (ADS)

    Schmidt, Christoph F.; Fakhri, Nikta; Battle, Christopher; Ott, Carolyn M.; Wessel, Alok D.; Lippincott-Schwartz, Jennifer; Mackintosh, Frederick C.

    2015-03-01

    Cells are active systems with molecular force generation that drives complex dynamics at the supramolecular scale. Much of cellular dynamics is driven by myosin motors interacting with the actin cytoskeleton. We discovered active random ``stirring'' driven by cytoplasmic myosin as an intermediate mode of transport, different from both thermal diffusion and directed motor activity. We found a further manifestation of cytoskeletal dynamics in the active motion patterns of primary cilia generated by epithelial cells. These cilia were thought to be immotile due to the absence of dynein motors, but it turns out that their anchoring deeper inside the cell in combination with the strongly fluctuating cortex results in clearly measurable non-equilibrium fluctuations.

  18. Galectin-7 modulates the length of the primary cilia and wound repair in polarized kidney epithelial cells.

    PubMed

    Rondanino, Christine; Poland, Paul A; Kinlough, Carol L; Li, Hui; Rbaibi, Youssef; Myerburg, Michael M; Al-bataineh, Mohammad M; Kashlan, Ossama B; Pastor-Soler, Nuria M; Hallows, Kenneth R; Weisz, Ora A; Apodaca, Gerard; Hughey, Rebecca P

    2011-09-01

    Galectins (Gal) are β-galactoside-binding proteins that function in epithelial development and homeostasis. An overlapping role for Gal-3 and Gal-7 in wound repair was reported in stratified epithelia. Although Gal-7 was thought absent in simple epithelia, it was reported in a proteomic analysis of cilia isolated from cultured human airway, and we recently identified Gal-7 transcripts in Madin-Darby canine kidney (MDCK) cells (Poland PA, Rondanino C, Kinlough CL, Heimburg-Molinaro J, Arthur CM, Stowell SR, Smith DF, Hughey RP. J Biol Chem 286: 6780-6790, 2011). We now report that Gal-7 is localized exclusively on the primary cilium of MDCK, LLC-PK(1) (pig kidney), and mpkCCD(c14) (mouse kidney) cells as well as on cilia in the rat renal proximal tubule. Gal-7 is also present on most cilia of multiciliated cells in human airway epithelia primary cultures. Interestingly, exogenous glutathione S-transferase (GST)-Gal-7 bound the MDCK apical plasma membrane as well as the cilium, while the lectin Ulex europeaus agglutinin, with glycan preferences similar to Gal-7, bound the basolateral plasma membrane as well as the cilium. In pull-down assays, β1-integrin isolated from either the basolateral or apical/cilia membranes of MDCK cells was similarly bound by GST-Gal-7. Selective localization of Gal-7 to cilia despite the presence of binding sites on all cell surfaces suggests that intracellular Gal-7 is specifically delivered to cilia rather than simply binding to surface glycoconjugates after generalized secretion. Moreover, depletion of Gal-7 using tetracycline-induced short-hairpin RNA in mpkCCD(c14) cells significantly reduced cilia length and slowed wound healing in a scratch assay. We conclude that Gal-7 is selectively targeted to cilia and plays a key role in surface stabilization of glycoconjugates responsible for integrating cilia function with epithelial repair. PMID:21677144

  19. Axonemal Positioning and Orientation in 3-D Space for Primary Cilia: What is Known, What is Assumed, and What Needs Clarification

    PubMed Central

    Farnum, Cornelia E.; Wilsman, Norman J.

    2012-01-01

    Two positional characteristics of the ciliary axoneme – its location on the plasma membrane as it emerges from the cell, and its orientation in three-dimensional space – are known to be critical for optimal function of actively motile cilia (including nodal cilia), as well as for modified cilia associated with special senses. However, these positional characteristics have not been analyzed to any significant extent for primary cilia. This review briefly summarizes the history of knowledge of these two positional characteristics across a wide spectrum of cilia, emphasizing their importance for proper function. Then the review focuses what is known about these same positional characteristics for primary cilia in all major tissue types where they have been reported. The review emphasizes major areas that would be productive for future research for understanding how positioning and 3-D orientation of primary cilia may be related to their hypothesized signaling roles within different cellular populations. PMID:22012592

  20. Mechanical signals promote osteogenic fate through a primary cilia-mediated mechanism.

    PubMed

    Chen, Julia C; Hoey, David A; Chua, Mardonn; Bellon, Raymond; Jacobs, Christopher R

    2016-04-01

    It has long been suspected, but never directly shown, that bone formed to accommodate an increase in mechanical loading is related to the creation of osteoblasts from skeletal stem cells. Indeed, biophysical stimuli potently regulate osteogenic lineage commitmentin vitro In this study, we transplanted bone marrow cells expressing green fluorescent protein, to enable lineage tracing, and subjected mice to a biophysical stimulus, to elicit a bone-forming response. We detected cells derived from transplanted progenitors embedded within the bone matrix near active bone-forming surfaces in response to loading, demonstrating for the first time, that mechanical signals enhance the homing and attachment of bone marrow cells to bone surfaces and the commitment to an osteogenic lineage of these cellsin vivo Furthermore, we used an inducible Cre/Lox recombination system to delete kinesin family member 3A (Kif3a), a gene that is essential for primary cilia formation, at will in transplanted cells and their progeny, regardless of which tissue may have incorporated them. Disruption of the mechanosensing organelle, the primary cilium in a progenitor population, significantly decreased the amount of bone formed in response to mechanical stimulation. The collective results of our study directly demonstrate that, in a novel experimental stem cell mechanobiology model, mechanical signals enhance osteogenic lineage commitmentin vivoand that the primary cilium contributes to this process.-Chen, J. C., Hoey, D. A., Chua, M., Bellon, R., Jacobs, C. R. Mechanical signals promote osteogenic fate through a primary cilia-mediated mechanism. PMID:26675708

  1. Neonatal seizures induced by pentylenetetrazol or kainic acid disrupt primary cilia growth on developing mouse cortical neurons.

    PubMed

    Parker, Alexander K; Le, Megan M; Smith, Tyler S; Hoang-Minh, Lan B; Atkinson, Eric W; Ugartemendia, George; Semple-Rowland, Susan; Coleman, Jason E; Sarkisian, Matthew R

    2016-08-01

    Neonatal or early-life seizures (ELS) are often associated with life-long neurophysiological, cognitive and behavioral deficits, but the underlying mechanisms contributing to these deficits remain poorly understood. Newborn, post-migratory cortical neurons sprout ciliary buds (procilia) that mature into primary cilia. Disruption of the growth or signaling capabilities of these cilia has been linked to atypical neurite outgrowth from neurons and abnormalities in neuronal circuitry. Here, we tested the hypothesis that generalized seizures induced by pentylenetetrazol (PTZ) or kainic acid (KA) during early postnatal development impair neuronal and/or glial ciliogenesis. Mice received PTZ (50 or 100mg/kg), KA (2mg/kg), or saline either once at birth (P0), or once daily from P0 to P4. Using immunohistochemistry and electron microscopy, the cilia of neurons and glia were examined at P7, P14, and P42. A total of 83 regions were analyzed, representing 13 unique neocortical and hippocampal regions. Neuronal cilia were identified by co-expression of NeuN and type 3 adenylyl cyclase (ACIII) or somatostatin receptor 3 (SSTR3), while glial cilia were identified by co-expression of GFAP, Arl13b, and gamma-tubulin. We found that PTZ exposure at either P0 or from P0 to P4 induced convulsive behavior, followed by acute and lasting effects on neuronal cilia lengths that varied depending on the cortical region, PTZ dose, injection frequency, and time post-PTZ. Both increases and decreases in neuronal cilia length were observed. No changes in the length of glial cilia were observed under any of the test conditions. Lastly, we found that a single KA seizure at P0 led to similar abnormalities in neuronal cilia lengths. Our results suggest that seizure(s) occurring during early stages of cortical development induce persistent and widespread changes in neuronal cilia length. Given the impact neuronal cilia have on neuronal differentiation, ELS-induced changes in ciliogenesis may

  2. Primary cilia function regulates the length of the embryonic trunk axis and urogenital field in mice.

    PubMed

    Wainwright, Elanor N; Svingen, Terje; Ng, Ee Ting; Wicking, Carol; Koopman, Peter

    2014-11-15

    The issues of whether and how some organs coordinate their size and shape with the blueprint of the embryo axis, while others appear to regulate their morphogenesis autonomously, remain poorly understood. Mutations in Ift144, encoding a component of the trafficking machinery of primary cilia assembly, result in a range of embryo patterning defects, affecting the limbs, skeleton and neural system. Here, we show that embryos of the mouse mutant Ift144(twt) develop gonads that are larger than wild-type. Investigation of the early patterning of the urogenital ridge revealed that the anterior-posterior domain of the gonad/mesonephros was extended at 10.5 dpc, with no change in the length of the metanephros. In XY embryos, this extension resulted in an increase in testis cord number. Moreover, we observed a concomitant extension of the trunk axis in both sexes, with no change in the length of the tail domain or somite number. Our findings support a model in which: (1) primary cilia regulate embryonic trunk elongation; (2) the length of the trunk axis determines the size of the urogenital ridges; and (3) the gonad domain is partitioned into a number of testis cords that depends on the available space, rather than being divided a predetermined number of times to generate a specific number of cords. PMID:25224227

  3. Phosphorylation-dependent Akt-Inversin interaction at the basal body of primary cilia.

    PubMed

    Suizu, Futoshi; Hirata, Noriyuki; Kimura, Kohki; Edamura, Tatsuma; Tanaka, Tsutomu; Ishigaki, Satoko; Donia, Thoria; Noguchi, Hiroko; Iwanaga, Toshihiko; Noguchi, Masayuki

    2016-06-15

    A primary cilium is a microtubule-based sensory organelle that plays an important role in human development and disease. However, regulation of Akt in cilia and its role in ciliary development has not been demonstrated. Using yeast two-hybrid screening, we demonstrate that Inversin (INVS) interacts with Akt. Mutation in the INVS gene causes nephronophthisis type II (NPHP2), an autosomal recessive chronic tubulointerstitial nephropathy. Co-immunoprecipitation assays show that Akt interacts with INVS via the C-terminus. In vitro kinase assays demonstrate that Akt phosphorylates INVS at amino acids 864-866 that are required not only for Akt interaction, but also for INVS dimerization. Co-localization of INVS and phosphorylated form of Akt at the basal body is augmented by PDGF-AA Akt-null MEF cells as well as siRNA-mediated inhibition of Akt attenuated ciliary growth, which was reversed by Akt reintroduction. Mutant phosphodead- or NPHP2-related truncated INVS, which lack Akt phosphorylation sites, suppress cell growth and exhibit distorted lumen formation and misalignment of spindle axis during cell division. Further studies will be required for elucidating functional interactions of Akt-INVS at the primary cilia for identifying the molecular mechanisms underlying NPHP2. PMID:27220846

  4. Silibinin negatively contributes to primary cilia length via autophagy regulated by histone deacetylase 6 in confluent mouse embryo fibroblast 3T3-L1 cells.

    PubMed

    Xu, Qian; Liu, Wei; Liu, Xiaoling; Liu, Weiwei; Wang, Hongju; Yao, Guodong; Zang, Linghe; Hayashi, Toshihiko; Tashiro, Shin-Ichi; Onodera, Satoshi; Ikejima, Takashi

    2016-09-01

    Primary cilium is a cellular antenna, signalling as a sensory organelle. Numerous pathological manifestation is associated with change of its length. Although the interaction between autophagy and primary cilia has been suggested, the role of autophagy in primary cilia length is largely unknown. In this study the primary cilia were immunostained and observed by using confocal fluorescence microscopy, and we found that silibinin, a natural flavonoid, shortened the length of primary cilia, meanwhile it also induced autophagy in 3T3-L1 cells. This study was designed to investigate the significance of silibinin-induced autophagy in primary ciliary structure in confluent mouse embryo fibroblast 3T3-L1 cells. Either blocking the autophagic flux with pre-treatment with the autophagy inhibitor, 3-methyladenine (3-MA), or transfection of siRNA targeting LC3 inhibited the reduction of cilia length caused by silibinin exposure. Autophagy induced by silibinin decreased expressions of the cilia-associated proteins, such as IFT88, KIF3a and Ac-tubulin, while 3-MA restored it, indicating that autophagy induced by silibinin led to a reduction of primary cilia length. Histone deacetylase 6 (HDAC6), which was suggested as a mediator of autophagy, was up-regulated by silibinin in a time-dependent manner. In addition, 3T3-L1 cells treated with siRNA against HDAC6 had a reduced autophagic level and were protected from silibinin-induced cilia shortening. Taken together, we conclude that the HDAC6-mediated autophagy negatively regulates primary cilia length during silibinin treatment and has the potential to serve as a therapeutic target for primary cilia-associated ciliopathies. These findings thus provide new information about the potential link between autophagy and primary cilia. PMID:27435857

  5. Meckel-Gruber syndrome and the role of primary cilia in kidney, skeleton, and central nervous system development

    PubMed Central

    Barker, Amy R; Thomas, Rhys; Dawe, Helen R

    2014-01-01

    The ciliopathies are a group of related inherited diseases characterized by malformations in organ development. The diseases affect multiple organ systems, with kidney, skeleton, and brain malformations frequently observed. Research over the last decade has revealed that these diseases are due to defects in primary cilia, essential sensory organelles found on most cells in the human body. Here we discuss the genetic and cell biological basis of one of the most severe ciliopathies, Meckel-Gruber syndrome, and explain how primary cilia contribute to the development of the affected organ systems. PMID:24322779

  6. Primary cilia distribution and orientation during involution of the bovine mammary gland.

    PubMed

    Biet, J; Poole, C A; Stelwagen, K; Margerison, J K; Singh, K

    2016-05-01

    The regulation of mammary gland involution occurs through multiple levels including environmental factors, hormones, and local intramammary signals. Primary cilia (PC) are signaling organelles that sense biochemical and biophysical extracellular stimuli and are vital for cellular and tissue function. The aim of this study was to examine the distribution, incidence, and orientation of PC. Furthermore, we determined changes in expression levels of the signal transducer and activator of transcription (STAT)6 at the onset of bovine mammary gland involution. Mammary tissue was collected from pasture-fed, primiparous, nonpregnant Friesian dairy cows at mid lactation (n=5 per group) killed 6-h after milking (lactating controls) and during involution after 7 and 28 d of nonmilking (NM). Fluorescent immunohistochemistry and confocal microscopy of tissue sections showed that PC were present on luminal secretory epithelial cells (SEC), myoepithelial cells (MEC), and stromal fibroblast cells (SFC). Furthermore, in all 3 experimental groups, different PC positions or orientations relative to the cell surface were identified on SEC and MEC, which projected toward the lumen and were either straight, bent, or deflected against the apical cell surface, whereas PC in SFC were confined to the interalveolar space. However, by 28-d NM, fewer PC projected into the luminal space and most appeared deflected or projected toward the interalveolar space. Furthermore, by 28-d NM, with the increase in stromal connective tissue, more PC were detected within the interalveolar and interlobular stroma. At 28-d NM, we observed a decrease in luminal cilia relative to the total number of cilia. The number of ciliated cells in the total fraction (SEC, MEC, and SFC) was the same for all 3 groups, although in the luminal fraction (SEC and MEC), PC per nuclei increased by 28-d NM relative to lactation. At all 3 stages, we detected variations in shape and orientation of PC within the same alveolus, with

  7. RTTN Mutations Link Primary Cilia Function to Organization of the Human Cerebral Cortex

    PubMed Central

    Kheradmand Kia, Sima; Verbeek, Elly; Engelen, Erik; Schot, Rachel; Poot, Raymond A.; de Coo, Irenaeus F.M.; Lequin, Maarten H.; Poulton, Cathryn J.; Pourfarzad, Farzin; Grosveld, Frank G.; Brehm, António; de Wit, Marie Claire Y.; Oegema, Renske; Dobyns, William B.; Verheijen, Frans W.; Mancini, Grazia M.S.

    2012-01-01

    Polymicrogyria is a malformation of the developing cerebral cortex caused by abnormal organization and characterized by many small gyri and fusion of the outer molecular layer. We have identified autosomal-recessive mutations in RTTN, encoding Rotatin, in individuals with bilateral diffuse polymicrogyria from two separate families. Rotatin determines early embryonic axial rotation, as well as anteroposterior and dorsoventral patterning in the mouse. Human Rotatin has recently been identified as a centrosome-associated protein. The Drosophila melanogaster homolog of Rotatin, Ana3, is needed for structural integrity of centrioles and basal bodies and maintenance of sensory neurons. We show that Rotatin colocalizes with the basal bodies at the primary cilium. Cultured fibroblasts from affected individuals have structural abnormalities of the cilia and exhibit downregulation of BMP4, WNT5A, and WNT2B, which are key regulators of cortical patterning and are expressed at the cortical hem, the cortex-organizing center that gives rise to Cajal-Retzius (CR) neurons. Interestingly, we have shown that in mouse embryos, Rotatin colocalizes with CR neurons at the subpial marginal zone. Knockdown experiments in human fibroblasts and neural stem cells confirm a role for RTTN in cilia structure and function. RTTN mutations therefore link aberrant ciliary function to abnormal development and organization of the cortex in human individuals. PMID:22939636

  8. Primary ciliary dyskinesia: evaluation using cilia beat frequency assessment via spectral analysis of digital microscopy images

    PubMed Central

    Kögler, João E.; Macchione, Mariangela; Shoemark, Amelia; Saldiva, Paulo H. N.; Rodrigues, Joaquim C.

    2011-01-01

    Ciliary beat frequency (CBF) measurements provide valuable information for diagnosing of primary ciliary dyskinesia (PCD). We developed a system for measuring CBF, used it in association with electron microscopy to diagnose PCD, and then analyzed characteristics of PCD patients.1 The CBF measurement system was based on power spectra measured through digital imaging. Twenty-four patients suspected of having PCD (age 1–19 yr) were selected from a group of 75 children and adolescents with pneumopathies of unknown causes. Ten healthy, nonsmoking volunteers (age ≥17 yr) served as a control group. Nasal brush samples were collected, and CBF and electron microscopy were performed. PCD was diagnosed in 12 patients: 5 had radial spoke defects, 3 showed absent central microtubule pairs with transposition, 2 had outer dynein arm defects, 1 had a shortened outer dynein arm, and 1 had a normal ultrastructure. Previous studies have reported that the most common cilia defects are in the dynein arm. As expected, the mean CBF was higher in the control group (P < 0.001) and patients with normal ultrastructure (P < 0.002), than in those diagnosed with cilia ultrastructural defects (i.e., PCD patients). An obstructive ventilatory pattern was observed in 70% of the PCD patients who underwent pulmonary function tests. All PCD patients presented bronchial wall thickening on chest computed tomography scans. The protocol and diagnostic techniques employed allowed us to diagnose PCD in 16% of patients in this study. PMID:21551013

  9. Single-molecule imaging of Hedgehog pathway protein Smoothened in primary cilia reveals binding events regulated by Patched1

    PubMed Central

    Milenkovic, Ljiljana; Weiss, Lucien E.; Yoon, Joshua; Roth, Theodore L.; Su, YouRong S.; Sahl, Steffen J.; Scott, Matthew P.; Moerner, W. E.

    2015-01-01

    Accumulation of the signaling protein Smoothened (Smo) in the membrane of primary cilia is an essential step in Hedgehog (Hh) signal transduction, yet the molecular mechanisms of Smo movement and localization are poorly understood. Using ultrasensitive single-molecule tracking with high spatial/temporal precision (30 nm/10 ms), we discovered that binding events disrupt the primarily diffusive movement of Smo in cilia at an array of sites near the base. The affinity of Smo for these binding sites was modulated by the Hh pathway activation state. Activation, by either a ligand or genetic loss of the negatively acting Hh receptor Patched-1 (Ptch), reduced the affinity and frequency of Smo binding at the base. Our findings quantify activation-dependent changes in Smo dynamics in cilia and highlight a previously unknown step in Hh pathway activation. PMID:26100903

  10. Primary cilia act as mechanosensors during bone healing around an implant.

    PubMed

    Leucht, P; Monica, S D; Temiyasathit, S; Lenton, K; Manu, A; Longaker, M T; Jacobs, C R; Spilker, R L; Guo, H; Brunski, J B; Helms, J A

    2013-03-01

    The primary cilium is an organelle that senses cues in a cell's local environment. Some of these cues constitute molecular signals; here, we investigate the extent to which primary cilia can also sense mechanical stimuli. We used a conditional approach to delete Kif3a in pre-osteoblasts and then employed a motion device that generated a spatial distribution of strain around an intra-osseous implant positioned in the mouse tibia. We correlated interfacial strain fields with cell behaviors ranging from proliferation through all stages of osteogenic differentiation. We found that peri-implant cells in the Col1Cre;Kif3a(fl/fl) mice were unable to proliferate in response to a mechanical stimulus, failed to deposit and then orient collagen fibers to the strain fields caused by implant displacement, and failed to differentiate into bone-forming osteoblasts. Collectively, these data demonstrate that the lack of a functioning primary cilium blunts the normal response of a cell to a defined mechanical stimulus. The ability to manipulate the genetic background of peri-implant cells within the context of a whole, living tissue provides a rare opportunity to explore mechanotransduction from a multi-scale perspective. PMID:22784673

  11. Primary cilia act as mechanosensors during bone healing around an implant

    PubMed Central

    Leucht, P.; Monica, S.D.; Temiyasathit, S.; Lenton, K.; Manu, A.; Longaker, M.T.; Jacobs, C.R.; Spilker, R.L.; Guo, H.; Brunski, J.B.; Helms, J.A.

    2012-01-01

    The primary cilium is an organelle that senses cues in a cell’s local environment. Some of these cues constitute molecular signals; here, we investigate the extent to which primary cilia can also sense mechanical stimuli. We used a conditional approach to delete Kif3a in pre-osteoblasts and then employed a motion device that generated a spatial distribution of strain around an intra-osseous implant positioned in the mouse tibia. We correlated interfacial strain fields with cell behaviors ranging from proliferation through all stages of osteogenic differentiation. We found that peri-implant cells in the Col1Cre;Kif3afl/fl mice were unable to proliferate in response to a mechanical stimulus, failed to deposit and then orient collagen fibers to the strain fields caused by implant displacement, and failed to differentiate into bone-forming osteoblasts. Collectively, these data demonstrate that the lack of a functioning primary cilium blunts the normal response of a cell to a defined mechanical stimulus. The ability to manipulate the genetic background of peri-implant cells within the context of a whole, living tissue provides a rare opportunity to explore mechanotransduction from a multi-scale perspective. PMID:22784673

  12. Biological and Chemical Removal of Primary Cilia Affects Mechanical Activation of Chondrogenesis Markers in Chondroprogenitors and Hypertrophic Chondrocytes

    PubMed Central

    Deren, Matthew E.; Yang, Xu; Guan, Yingjie; Chen, Qian

    2016-01-01

    Chondroprogenitors and hypertrophic chondrocytes, which are the first and last stages of the chondrocyte differentiation process, respectively, are sensitive to mechanical signals. We hypothesize that the mechanical sensitivity of these cells depends on the cell surface primary cilia. To test this hypothesis, we removed the primary cilia by biological means with transfection with intraflagellar transport protein 88 (IFT88) siRNA or by chemical means with chloral hydrate treatment. Transfection of IFT88 siRNA significantly reduced the percentage of ciliated cells in both chondroprogenitor ATDC5 cells as well as primary hypertrophic chondrocytes. Cyclic loading (1 Hz, 10% matrix deformation) of ATDC5 cells in three-dimensional (3D) culture stimulates the mRNA levels of chondrogenesis marker Type II collagen (Col II), hypertrophic chondrocyte marker Type X collagen (Col X), and a molecular regulator of chondrogenesis and chondrocyte hypertrophy bone morphogenetic protein 2 (BMP-2). The reduction of ciliated chondroprogenitors abolishes mechanical stimulation of Col II, Col X, and BMP-2. In contrast, cyclic loading stimulates Col X mRNA levels in hypertrophic chondrocytes, but not those of Col II and BMP-2. Both biological and chemical reduction of ciliated hypertrophic chondrocytes reduced but failed to abolish mechanical stimulation of Col X mRNA levels. Thus, primary cilia play a major role in mechanical stimulation of chondrogenesis and chondrocyte hypertrophy in chondroprogenitor cells and at least a partial role in hypertrophic chondrocytes. PMID:26861287

  13. Primary cilia regulate mTORC1 activity and cell size through Lkb1.

    PubMed

    Boehlke, Christopher; Kotsis, Fruzsina; Patel, Vishal; Braeg, Simone; Voelker, Henriette; Bredt, Saskia; Beyer, Theresa; Janusch, Heike; Hamann, Christoph; Gödel, Markus; Müller, Klaus; Herbst, Martin; Hornung, Miriam; Doerken, Mara; Köttgen, Michael; Nitschke, Roland; Igarashi, Peter; Walz, Gerd; Kuehn, E Wolfgang

    2010-11-01

    The mTOR pathway is the central regulator of cell size. External signals from growth factors and nutrients converge on the mTORC1 multi-protein complex to modulate downstream targets, but how the different inputs are integrated and translated into specific cellular responses is incompletely understood. Deregulation of the mTOR pathway occurs in polycystic kidney disease (PKD), where cilia (filiform sensory organelles) fail to sense urine flow because of inherited mutations in ciliary proteins. We therefore investigated if cilia have a role in mTOR regulation. Here, we show that ablation of cilia in transgenic mice results in enlarged cells when compared with control animals. In vitro analysis demonstrated that bending of the cilia by flow is required for mTOR downregulation and cell-size control. Surprisingly, regulation of cell size by cilia is independent of flow-induced calcium transients, or Akt. However, the tumour-suppressor protein Lkb1 localises in the cilium, and flow results in increased AMPK phosphorylation at the basal body. Conversely, knockdown of Lkb1 prevents normal cell-size regulation under flow conditions. Our results demonstrate that the cilium regulates mTOR signalling and cell size, and identify the cilium-basal body compartment as a spatially restricted activation site for Lkb1 signalling. PMID:20972424

  14. Functional aspects of primary cilia in signaling, cell cycle and tumorigenesis

    PubMed Central

    2013-01-01

    Dysfunctional cilia underlie a broad range of cellular and tissue phenotypes and can eventually result in the development of ciliopathies: pathologically diverse diseases that range from clinically mild to highly complex and severe multi-organ failure syndromes incompatible with neonatal life. Given that virtually all cells of the human body have the capacity to generate cilia, it is likely that clinical manifestations attributed to ciliary dysfunction will increase in the years to come. Disputed but nevertheless enigmatic is the notion that at least a subset of tumor phenotypes fit within the ciliopathy disease spectrum and that cilia loss may be required for tumor progression. Contending for the centrosome renders ciliation and cell division mutually exclusive; a regulated tipping of balance promotes either process. The mechanisms involved, however, are complex. If the hypothesis that tumorigenesis results from dysfunctional cilia is true, then why do the classic ciliopathies only show limited hyperplasia at best? Although disassembly of the cilium is a prerequisite for cell proliferation, it does not intrinsically drive tumorigenesis per se. Alternatively, we will explore the emerging evidence suggesting that some tumors depend on ciliary signaling. After reviewing the structure, genesis and signaling of cilia, the various ciliopathy syndromes and their genetics, we discuss the current debate of tumorigenesis as a ciliopathy spectrum defect, and describe recent advances in this fascinating field. PMID:23628112

  15. Kinetics of Hedgehog-Dependent Full-Length Gli3 Accumulation in Primary Cilia and Subsequent Degradation ▿ † ‡

    PubMed Central

    Wen, Xiaohui; Lai, Cary K.; Evangelista, Marie; Hongo, Jo-Anne; de Sauvage, Frederic J.; Scales, Suzie J.

    2010-01-01

    Hedgehog (Hh) signaling in vertebrates depends on intraflagellar transport (IFT) within primary cilia. The Hh receptor Patched is found in cilia in the absence of Hh and is replaced by the signal transducer Smoothened within an hour of Hh stimulation. By generating antibodies capable of detecting endogenous pathway transcription factors Gli2 and Gli3, we monitored their kinetics of accumulation in cilia upon Hh stimulation. Localization occurs within minutes of Hh addition, making it the fastest reported readout of pathway activity, which permits more precise temporal and spatial localization of Hh signaling events. We show that the species of Gli3 that accumulates at cilium tips is full-length and likely not protein kinase A phosphorylated. We also confirmed that phosphorylation and βTrCP/Cul1 are required for endogenous Gli3 processing and that this is inhibited by Hh. Surprisingly, however, Hh-dependent inhibition of processing does not lead to accumulation of full-length Gli3, but instead renders it labile, leading to its proteasomal degradation via the SPOP/Cul3 complex. In fact, full-length Gli3 disappears with faster kinetics than the Gli3 repressor, the latter not requiring SPOP/Cul3 or βTrCP/Cul1. This may contribute to the increased Gli3 activator/repressor ratios found in IFT mutants. PMID:20154143

  16. EFCAB7 and IQCE regulate Hedgehog signaling by tethering the EVC-EVC2 complex to the base of primary cilia

    PubMed Central

    Pusapati, Ganesh V.; Hughes, Casey E; Dorn, Karolin V.; Zhang, Dapeng; Sugianto, Priscilla; Aravind, L.; Rohatgi, Rajat

    2014-01-01

    The Hedgehog (Hh) pathway depends on primary cilia in vertebrates, but the signaling machinery within cilia remains incompletely defined. We report the identification of a complex between two ciliary proteins, EFCAB7 and IQCE, which positively regulates the Hh pathway. The EFCAB7-IQCE module anchors the EVC-EVC2 complex in a signaling microdomain at the base of cilia. EVC and EVC2 genes are mutated in Ellis van Creveld and Weyers syndromes, characterized by impaired Hh signaling in skeletal, cardiac and orofacial tissues. EFCAB7 binds to a C-terminal disordered region in EVC2 that is deleted in Weyers patients. EFCAB7 depletion mimics the Weyers cellular phenotype— the mis-localization of EVC-EVC2 within cilia and impaired activation of the transcription factor GLI2. Evolutionary analysis suggests that emergence of these complexes might have been important for adaptation of an ancient organelle, the cilium, for an animal-specific signaling network. PMID:24582806

  17. Concise Review: Primary Cilia: Control Centers for Stem Cell Lineage Specification and Potential Targets for Cell-Based Therapies.

    PubMed

    Bodle, Josephine C; Loboa, Elizabeth G

    2016-06-01

    Directing stem cell lineage commitment prevails as the holy grail of translational stem cell research, particularly to those interested in the application of mesenchymal stem cells and adipose-derived stem cells in tissue engineering. However, elucidating the mechanisms underlying their phenotypic specification persists as an active area of research. In recent studies, the primary cilium structure has been intimately associated with defining cell phenotype, maintaining stemness, as well as functioning in a chemo, electro, and mechanosensory capacity in progenitor and committed cell types. Many hypothesize that the primary cilium may indeed be another important player in defining and controlling cell phenotype, concomitant with lineage-dictated cytoskeletal dynamics. Many of the studies on the primary cilium have emerged from disparate areas of biological research, and crosstalk amongst these areas of research is just beginning. To date, there has not been a thorough review of how primary cilia fit into the current paradigm of stem cell differentiation and this review aims to summarize the current cilia work in this context. The goal of this review is to highlight the cilium's function and integrate this knowledge into the working knowledge of stem cell biologists and tissue engineers developing regenerative medicine technologies. Stem Cells 2016;34:1445-1454. PMID:26866419

  18. Inv acts as a molecular anchor for Nphp3 and Nek8 in the proximal segment of primary cilia.

    PubMed

    Shiba, Dai; Manning, Danielle K; Koga, Hisashi; Beier, David R; Yokoyama, Takahiko

    2010-02-01

    A primary cilium is an antenna-like structure extending from the surface of most vertebrate cells. It is structurally divided along its vertical axis into sub-compartments that include the ciliary tip, the shaft, the ciliary necklace segment, the transitional zone and the basal body. We recently discovered that the shaft of the primary cilia has a distinct molecular compartment, termed the "Inv compartment", which is characterized by the accumulation of Inv at the base of primary cilia. Inv was discovered as a causative gene in inv mutant mice. It was later found to be responsible for the infantile type of nephronophthisis (NPHP2). Nephronophthisis (NPHP) is an autosomal recessive kidney disease. Nine causative genes have been identified, with all examined products thought to function in cilia, basal body and/or centrioles. However, their exact intra-ciliary localization and relationship have not been clear. Here, we report that products of Nphp3 and Nek8 (the mouse orthologs of the causative genes for NPHP3 and NPHP9, respectively) localize to the Inv compartment. We also show that Inv is essential for the compartmental localization of Nphp3 and Nek8, whereas localization of Inv does not require Nphp3 or Nek8. Nphp1 and Nphp4 also localize at the proximal region of the cilium, but not in Inv compartment. Our results indicate that Inv acts as an anchor for Nphp3 and Nek8 in the Inv compartment, and suggest that Inv compartment is a candidate site for intra-ciliary interaction of Inv, Nphp3 and Nek8. PMID:20169535

  19. Shear stress blunts tubuloglomerular feedback partially mediated by primary cilia and nitric oxide at the macula densa.

    PubMed

    Wang, Lei; Shen, Chunyu; Liu, Haifeng; Wang, Shaohui; Chen, Xinshan; Roman, Richard J; Juncos, Luis A; Lu, Yan; Wei, Jin; Zhang, Jie; Yip, Kay-Pong; Liu, Ruisheng

    2015-10-01

    The present study tested whether primary cilia on macula densa serve as a flow sensor to enhance nitric oxide synthase 1 (NOS1) activity and inhibit tubuloglomerular feedback (TGF). Isolated perfused macula densa was loaded with calcein red and 4,5-diaminofluorescein diacetate to monitor cell volume and nitric oxide (NO) generation. An increase in tubular flow rate from 0 to 40 nl/min enhanced NO production by 40.0 ± 1.2%. The flow-induced NO generation was blocked by an inhibitor of NOS1 but not by inhibition of the Na/K/2Cl cotransporter or the removal of electrolytes from the perfusate. NO generation increased from 174.8 ± 21 to 276.1 ± 24 units/min in cultured MMDD1 cells when shear stress was increased from 0.5 to 5.0 dynes/cm(2). The shear stress-induced NO generation was abolished in MMDD1 cells in which the cilia were disrupted using a siRNA to ift88. Increasing the NaCl concentration of the tubular perfusate from 10 to 80 mM NaCl in the isolated perfused juxtaglomerular preparation reduced the diameter of the afferent arteriole by 3.8 ± 0.1 μm. This response was significantly blunted to 2.5 ± 0.2 μm when dextran was added to the perfusate to increase the viscosity and shear stress. Inhibition of NOS1 blocked the effect of dextran on TGF response. In vitro, the effects of raising perfusate viscosity with dextran on tubular hydraulic pressure were minimized by reducing the outflow resistance to avoid stretching of tubular cells. These results suggest that shear stress stimulates primary cilia on the macula densa to enhance NO generation and inhibit TGF responsiveness. PMID:26269519

  20. Planar cell polarity effector gene Intu regulates cell fate-specific differentiation of keratinocytes through the primary cilia.

    PubMed

    Dai, D; Li, L; Huebner, A; Zeng, H; Guevara, E; Claypool, D J; Liu, A; Chen, J

    2013-01-01

    Genes involved in the planar cell polarity (PCP) signaling pathway are essential for a number of developmental processes in mammals, such as convergent extension and ciliogenesis. Tissue-specific PCP effector genes of the PCP signaling pathway are believed to mediate PCP signals in a tissue- and cell type-specific manner. However, how PCP signaling controls the morphogenesis of mammalian tissues remains unclear. In this study, we investigated the role of inturned (Intu), a tissue-specific PCP effector gene, during hair follicle formation in mice. Tissue-specific disruption of Intu in embryonic epidermis resulted in hair follicle morphogenesis arrest because of the failure of follicular keratinocyte to differentiate. Targeting Intu in the epidermis resulted in almost complete loss of primary cilia in epidermal and follicular keratinocytes, and a suppressed hedgehog signaling pathway. Surprisingly, the epidermal stratification and differentiation programs and barrier function were not affected. These results demonstrate that tissue-specific PCP effector genes of the PCP signaling pathway control the differentiation of keratinocytes through the primary cilia in a cell fate- and context-dependent manner, which may be critical in orchestrating the propagation and interpretation of polarity signals established by the core PCP components. PMID:22935613

  1. Changes in cell surface primary cilia and microvilli concurrent with measurements of fluid flow across the rabbit corneal endothelium ex vivo.

    PubMed

    Doughty, M J

    1998-12-01

    Primary cilia and microvilli have been reported on the mammalian rabbit corneal endothelium but their relationship to cell function is undefined. Six corneas from healthy 2 kg female albino rabbits were glutaraldehyde-fixed post mortem (15:00 h) or twelve corneal stroma-endothelial preparations incubated at 37 degrees C under an applied hydrostatic pressure of 20 cm H2O for 4 h prior to fixation. The corneal endothelium was assessed by quantitative scanning electron microscopy. Cells fixed immediately post mortem were decorated with small stubby microvilli (average 21 +/- 13/100 micron 2), and only 25% of the cells were decorated with primary cilia having an average length of 2.44 +/- 1.56 microns. Following 4 h ex vivo incubation with a phosphate-buffered Ringer solution, conspicuous microvilli developed to an average density of 40 +/- 19/100 micron 2 and primary cilia were found on 12% of the cells, having on average length of 2.27 +/- 1.38 microns. Following 4 h incubation in a bicarbonate-buffered Ringer solution, small stubby microvilli developed to a density of 49 +/- 18/100 micron 2, and 40% of the cells showed primary cilia with an average length of 4.31 +/- 1.93 microns; the net trans-endothelial fluid flow in the latter set was 60% greater. These studies indicate that the primary cilia on corneal endothelial cells might be responsive to fluid flow, but that mild mechanical and/or chemical stress could also be the cause of the change since the elaboration of primary cilia can be accompanied by microvilli as well. PMID:10036788

  2. Cilia Dysfunction in Lung Disease

    PubMed Central

    Tilley, Ann E.; Walters, Matthew S.; Shaykhiev, Renat; Crystal, Ronald G.

    2015-01-01

    A characteristic feature of the human airway epithelium is the presence of ciliated cells bearing motile cilia, specialized cell surface projections containing axonemes comprised of microtubules and dynein arms, which provide ATP-driven motility. In the airways, cilia function in concert with airway mucus to mediate the critical function of mucociliary clearance, cleansing the airways of inhaled particles and pathogens. The prototypical disorder of respiratory cilia is primary ciliary dyskinesia, an inherited disorder that leads to impaired mucociliary clearance, repeated chest infections, and progressive destruction of lung architecture. Numerous acquired lung diseases are also marked by abnormalities in both cilia structure and function. In this review we summarize current knowledge regarding airway ciliated cells and cilia, how they function to maintain a healthy epithelium, and how disorders of cilia structure and function contribute to inherited and acquired lung disease. PMID:25386990

  3. Cilia dysfunction in lung disease.

    PubMed

    Tilley, Ann E; Walters, Matthew S; Shaykhiev, Renat; Crystal, Ronald G

    2015-01-01

    A characteristic feature of the human airway epithelium is the presence of ciliated cells bearing motile cilia, specialized cell surface projections containing axonemes composed of microtubules and dynein arms, which provide ATP-driven motility. In the airways, cilia function in concert with airway mucus to mediate the critical function of mucociliary clearance, cleansing the airways of inhaled particles and pathogens. The prototypical disorder of respiratory cilia is primary ciliary dyskinesia, an inherited disorder that leads to impaired mucociliary clearance, to repeated chest infections, and to the progressive destruction of lung architecture. Numerous acquired lung diseases are also marked by abnormalities in both cilia structure and function. In this review we summarize current knowledge regarding airway ciliated cells and cilia, how they function to maintain a healthy epithelium, and how disorders of cilia structure and function contribute to inherited and acquired lung disease. PMID:25386990

  4. PTEN regulates cilia through Dishevelled

    PubMed Central

    Shnitsar, Iryna; Bashkurov, Mikhail; Masson, Glenn R.; Ogunjimi, Abiodun A.; Mosessian, Sherly; Cabeza, Eduardo Aguiar; Hirsch, Calley L.; Trcka, Daniel; Gish, Gerald; Jiao, Jing; Wu, Hong; Winklbauer, Rudolf; Williams, Roger L.; Pelletier, Laurence; Wrana, Jeffrey L.; Barrios-Rodiles, Miriam

    2015-01-01

    Cilia are hair-like cellular protrusions important in many aspects of eukaryotic biology. For instance, motile cilia enable fluid movement over epithelial surfaces, while primary (sensory) cilia play roles in cellular signalling. The molecular events underlying cilia dynamics, and particularly their disassembly, are not well understood. Phosphatase and tensin homologue (PTEN) is an extensively studied tumour suppressor, thought to primarily act by antagonizing PI3-kinase signalling. Here we demonstrate that PTEN plays an important role in multicilia formation and cilia disassembly by controlling the phosphorylation of Dishevelled (DVL), another ciliogenesis regulator. DVL is a central component of WNT signalling that plays a role during convergent extension movements, which we show here are also regulated by PTEN. Our studies identify a novel protein substrate for PTEN that couples PTEN to regulation of cilia dynamics and WNT signalling, thus advancing our understanding of potential underlying molecular etiologies of PTEN-related pathologies. PMID:26399523

  5. NIMA-related kinases defective in murine models of polycystic kidney diseases localize to primary cilia and centrosomes.

    PubMed

    Mahjoub, Moe R; Trapp, Melissa L; Quarmby, Lynne M

    2005-12-01

    A key feature of the polycystic kidney diseases is aberrant cell proliferation, a consequence of dysfunctional ciliary signaling. The NIMA-related kinases (Nek) Nek1 and Nek8 carry the causal mutations of two of the eight established mouse models of polycystic kidneys. Nek proteins have roles in cell cycle and may contribute to coordinate regulation of cilia and cell-cycle progression. Herein is reported that in a mouse kidney epithelial cell line, mNek1 localizes to centrosomes in interphase and remains associated with the mitotic spindle pole during mitosis. In contrast, mNek8 localizes to the proximal region of the primary cilium and is not observed in dividing cells. Knockdown of mNek8 by siRNA does not affect ciliary assembly. Taken together with the phenotypes of the mutant mice, these data suggest that mNek1 and mNek8 provide links between cilia, centrosomes, and cell-cycle regulation. PMID:16267153

  6. Over-expression of Plk4 induces centrosome amplification, loss of primary cilia and associated tissue hyperplasia in the mouse

    PubMed Central

    Coelho, Paula A.; Bury, Leah; Shahbazi, Marta N.; Liakath-Ali, Kifayathullah; Tate, Peri H.; Wormald, Sam; Hindley, Christopher J.; Huch, Meritxell; Archer, Joy; Skarnes, William C.; Zernicka-Goetz, Magdalena; Glover, David M.

    2015-01-01

    To address the long-known relationship between supernumerary centrosomes and cancer, we have generated a transgenic mouse that permits inducible expression of the master regulator of centriole duplication, Polo-like-kinase-4 (Plk4). Over-expression of Plk4 from this transgene advances the onset of tumour formation that occurs in the absence of the tumour suppressor p53. Plk4 over-expression also leads to hyperproliferation of cells in the pancreas and skin that is enhanced in a p53 null background. Pancreatic islets become enlarged following Plk4 over-expression as a result of equal expansion of α- and β-cells, which exhibit centrosome amplification. Mice overexpressing Plk4 develop grey hair due to a loss of differentiated melanocytes and bald patches of skin associated with a thickening of the epidermis. This reflects an increase in proliferating cells expressing keratin 5 in the basal epidermal layer and the expansion of these cells into suprabasal layers. Such cells also express keratin 6, a marker for hyperplasia. This is paralleled by a decreased expression of later differentiation markers, involucrin, filaggrin and loricrin. Proliferating cells showed an increase in centrosome number and a loss of primary cilia, events that were mirrored in primary cultures of keratinocytes established from these animals. We discuss how repeated duplication of centrioles appears to prevent the formation of basal bodies leading to loss of primary cilia, disruption of signalling and thereby aberrant differentiation of cells within the epidermis. The absence of p53 permits cells with increased centrosomes to continue dividing, thus setting up a neoplastic state of error prone mitoses, a prerequisite for cancer development. PMID:26701933

  7. Over-expression of Plk4 induces centrosome amplification, loss of primary cilia and associated tissue hyperplasia in the mouse.

    PubMed

    Coelho, Paula A; Bury, Leah; Shahbazi, Marta N; Liakath-Ali, Kifayathullah; Tate, Peri H; Wormald, Sam; Hindley, Christopher J; Huch, Meritxell; Archer, Joy; Skarnes, William C; Zernicka-Goetz, Magdalena; Glover, David M

    2015-12-01

    To address the long-known relationship between supernumerary centrosomes and cancer, we have generated a transgenic mouse that permits inducible expression of the master regulator of centriole duplication, Polo-like-kinase-4 (Plk4). Over-expression of Plk4 from this transgene advances the onset of tumour formation that occurs in the absence of the tumour suppressor p53. Plk4 over-expression also leads to hyperproliferation of cells in the pancreas and skin that is enhanced in a p53 null background. Pancreatic islets become enlarged following Plk4 over-expression as a result of equal expansion of α- and β-cells, which exhibit centrosome amplification. Mice overexpressing Plk4 develop grey hair due to a loss of differentiated melanocytes and bald patches of skin associated with a thickening of the epidermis. This reflects an increase in proliferating cells expressing keratin 5 in the basal epidermal layer and the expansion of these cells into suprabasal layers. Such cells also express keratin 6, a marker for hyperplasia. This is paralleled by a decreased expression of later differentiation markers, involucrin, filaggrin and loricrin. Proliferating cells showed an increase in centrosome number and a loss of primary cilia, events that were mirrored in primary cultures of keratinocytes established from these animals. We discuss how repeated duplication of centrioles appears to prevent the formation of basal bodies leading to loss of primary cilia, disruption of signalling and thereby aberrant differentiation of cells within the epidermis. The absence of p53 permits cells with increased centrosomes to continue dividing, thus setting up a neoplastic state of error prone mitoses, a prerequisite for cancer development. PMID:26701933

  8. Mutations in INVS encoding inversin cause nephronophthisis type 2, linking renal cystic disease to the function of primary cilia and left-right axis determination

    PubMed Central

    Otto, Edgar A; Schermer, Bernhard; Obara, Tomoko; O'Toole, John F; Hiller, Karl S; Mueller, Adelheid M; Ruf, Rainer G; Hoefele, Julia; Beekmann, Frank; Landau, Daniel; Foreman, John W; Goodship, Judith A; Strachan, Tom; Kispert, Andreas; Wolf, Matthias T; Gagnadoux, Marie F; Nivet, Hubert; Antignac, Corinne; Walz, Gerd; Drummond, Iain A; Benzing, Thomas; Hildebrandt, Friedhelm

    2013-01-01

    Nephronophthisis (NPHP), an autosomal recessive cystic kidney disease, leads to chronic renal failure in children. The genes mutated in NPHP1 and NPHP4 have been identified, and a gene locus associated with infantile nephronophthisis (NPHP2) was mapped. The kidney phenotype of NPHP2 combines clinical features of NPHP and polycystic kidney disease (PKD). Here, we identify inversin (INVS) as the gene mutated in NPHP2 with and without situs inversus. We show molecular interaction of inversin with nephrocystin, the product of the gene mutated in NPHP1 and interaction of nephrocystin with β-tubulin, a main component of primary cilia. We show that nephrocystin, inversin and β-tubulin colocalize to primary cilia of renal tubular cells. Furthermore, we produce a PKD-like renal cystic phenotype and randomization of heart looping by knockdown of invs expression in zebrafish. The interaction and colocalization in cilia of inversin, nephrocystin and β-tubulin connect pathogenetic aspects of NPHP to PKD, to primary cilia function and to left-right axis determination. PMID:12872123

  9. Cilia and Diseases

    PubMed Central

    Brown, Jason M.; Witman, George B.

    2014-01-01

    In recent decades, cilia have moved from relative obscurity to a position of importance for understanding multiple complex human diseases. Now termed the ciliopathies, these diseases inflict devastating effects on millions of people worldwide. In this review, written primarily for teachers and students who may not yet be aware of the recent exciting developments in this field, we provide a general overview of our current understanding of cilia and human disease. We start with an introduction to cilia structure and assembly and indicate where they are found in the human body. We then discuss the clinical features of selected ciliopathies, with an emphasis on primary ciliary dyskinesia, polycystic kidney disease, and retinal degeneration. The history of ciliopathy research involves a fascinating interplay between basic and clinical sciences, highlighted in a timeline. Finally, we summarize the relative strengths of individual model organisms for ciliopathy research; many of these are suitable for classroom use. PMID:25960570

  10. Revealing the Molecular Structure and the Transport Mechanism at the Base of Primary Cilia Using Superresolution STED Microscopy

    NASA Astrophysics Data System (ADS)

    Yang, Tung-Lin

    The primary cilium is an organelle that serves as a signaling center of the cell and is involved in the hedgehog signaling, cAMP pathway, Wnt pathways, etc. Ciliary function relies on the transportation of molecules between the primary cilium and the cell, which is facilitated by intraflagellar transport (IFT). IFT88, one of the important IFT proteins in complex B, is known to play a role in the formation and maintenance of cilia in various types of organisms. The ciliary transition zone (TZ), which is part of the gating apparatus at the ciliary base, is home to a large number of ciliopathy molecules. Recent studies have identified important regulating elements for TZ gating in cilia. However, the architecture of the TZ region and its arrangement relative to intraflagellar transport (IFT) proteins remain largely unknown, hindering the mechanistic understanding of the regulation processes. One of the major challenges comes from the tiny volume at the ciliary base packed with numerous proteins, with the diameter of the TZ close to the diffraction limit of conventional microscopes. Using a series of stimulated emission depletion (STED) superresolution images mapped to electron microscopy images, we analyzed the structural organization of the ciliary base. Subdiffraction imaging of TZ components defines novel geometric distributions of RPGRIP1L, MKS1, CEP290, TCTN2 and TMEM67, shedding light on their roles in TZ structure, assembly, and function. We found TCTN2 at the outmost periphery of the TZ close to the ciliary membrane, with a 227+/-18 nm diameter. TMEM67 was adjacent to TCTN2, with a 205+/-20 nm diameter. RPGRIP1L was localized toward the axoneme at the same axial level as TCTN2 and TMEM67, with a 165+/-8 nm diameter. MKS1 was situated between TMEM67 and RPGRIP1L, with an 186+/-21 nm diameter. Surprisingly, CEP290 was localized at the proximal side of the TZ close to the distal end of the centrin-labeled basal body. The lateral width was unexpectedly close to

  11. Morphogenesis of respiratory syncytial virus in human primary nasal ciliated epithelial cells occurs at surface membrane microdomains that are distinct from cilia

    SciTech Connect

    Jumat, Muhammad Raihan; Yan, Yan; Ravi, Laxmi Iyer; Wong, Puisan; Huong, Tra Nguyen; Li, Chunwei; Tan, Boon Huan; Wang, De Yun; Sugrue, Richard J.

    2015-10-15

    The distribution of cilia and the respiratory syncytial virus (RSV) nucleocapsid (N) protein, fusion (F) protein, attachment (G) protein, and M2-1 protein in human ciliated nasal epithelial cells was examined at between 1 and 5 days post-infection (dpi). All virus structural proteins were localized at cell surface projections that were distinct from cilia. The F protein was also trafficked into the cilia, and while its presence increased as the infection proceeded, the N protein was not detected in the cilia at any time of infection. The presence of the F protein in the cilia correlated with cellular changes in the cilia and reduced cilia function. At 5 dpi extensive cilia loss and further reduced cilia function was noted. These data suggested that although RSV morphogenesis occurs at non-cilia locations on ciliated nasal epithelial cells, RSV infection induces changes in the cilia body that leads to extensive cilia loss. - Highlights: • Respiratory syncytial virus (RSV) infects nasal ciliated epithelial cells. • Virus morphogenesis occurs within filamentous projections distinct from cilia. • The RSV N protein was not detected in the cilia at any time during infection. • Trafficking of the F protein into the cilia occurred early in infection. • Presence of the F protein in cilia correlated with impaired cilia function.

  12. Motile Cilia of Human Airway Epithelia Are Chemosensory

    PubMed Central

    Shah, Alok S; Ben-Shahar, Yehuda; Moninger, Thomas O; Kline, Joel N; Welsh, Michael J

    2010-01-01

    Cilia are microscopic projections that extend from eukaryotic cells. There are two general types of cilia; primary cilia serve as sensory organelles, whereas motile cilia exert mechanical force. The motile cilia emerging from human airway epithelial cells propel harmful inhaled material out of the lung. We found that these cells express sensory bitter taste receptors, which localized on motile cilia. Bitter compounds increased the intracellular Ca2+ concentration and stimulated ciliary beat frequency. Thus, airway epithelia contain a cell-autonomous system in which motile cilia both sense noxious substances entering airways and initiate a defensive mechanical mechanism to eliminate the offending compound. Hence, like primary cilia, classical motile cilia also contain sensors to detect the external environment. PMID:19628819

  13. Bone cell mechanosensation of fluid flow stimulation: a fluid-structure interaction model characterising the role integrin attachments and primary cilia.

    PubMed

    Vaughan, T J; Mullen, C A; Verbruggen, S W; McNamara, L M

    2015-08-01

    Load-induced fluid flow acts as an important biophysical signal for bone cell mechanotransduction in vivo, where the mechanical environment is thought to be monitored by integrin and primary cilia mechanoreceptors on the cell body. However, precisely how integrin- and primary cilia-based mechanosensors interact with the surrounding fluid flow stimulus and ultimately contribute to the biochemical response of bone cells within either the in vitro or in vivo environment remains poorly understood. In this study, we developed fluid-structure interaction models to characterise the deformation of integrin- and primary cilia-based mechanosensors in bone cells under fluid flow stimulation. Under in vitro fluid flow stimulation, these models predicted that integrin attachments on the cell-substrate interface were highly stimulated ε(eq) > 200,000 με, while the presence of a primary cilium on the cell also resulted in significant strain amplifications, arising at the ciliary base. As such, these mechanosensors likely play a role in mediating bone mechanotransduction in vitro. Under in vivo fluid flow stimulation, integrin attachments along the canalicular wall were highly stimulated and likely play a role in mediating cellular responses in vivo. The role of the primary cilium as a flow sensor in vivo depended upon its configuration within the lacunar cavity. Specifically, our results showed that a short free-standing primary cilium could not effectively fulfil a flow sensing role in vivo. However, a primary cilium that discretely attaches the lacunar wall can be highly stimulated, due to hydrodynamic pressure in the lacunocanalicular system and, as such, could play a role in mediating bone mechanotransduction in vivo. PMID:25399300

  14. The role of primary cilia in the development and disease of the retina

    PubMed Central

    Wheway, Gabrielle; Parry, David A; Johnson, Colin A

    2014-01-01

    The normal development and function of photoreceptors is essential for eye health and visual acuity in vertebrates. Mutations in genes encoding proteins involved in photoreceptor development and function are associated with a suite of inherited retinal dystrophies, often as part of complex multi-organ syndromic conditions. In this review, we focus on the role of the photoreceptor outer segment, a highly modified and specialized primary cilium, in retinal health and disease. We discuss the many defects in the structure and function of the photoreceptor primary cilium that can cause a class of inherited conditions known as ciliopathies, often characterized by retinal dystrophy and degeneration, and highlight the recent insights into disease mechanisms. PMID:24162842

  15. Dopamine receptors reveal an essential role of IFT-B, KIF17, and Rab23 in delivering specific receptors to primary cilia

    PubMed Central

    Leaf, Alison; Von Zastrow, Mark

    2015-01-01

    Appropriate physiological signaling by primary cilia depends on the specific targeting of particular receptors to the ciliary membrane, but how this occurs remains poorly understood. In this study, we show that D1-type dopaminergic receptors are delivered to cilia from the extra-ciliary plasma membrane by a mechanism requiring the receptor cytoplasmic tail, the intraflagellar transport complex-B (IFT-B), and ciliary kinesin KIF17. This targeting mechanism critically depends on Rab23, a small guanine nucleotide binding protein that has important effects on physiological signaling from cilia but was not known previously to be essential for ciliary delivery of any cargo. Depleting Rab23 prevents dopamine receptors from accessing the ciliary membrane. Conversely, fusion of Rab23 to a non-ciliary receptor is sufficient to drive robust, nucleotide-dependent mis-localization to the ciliary membrane. Dopamine receptors thus reveal a previously unrecognized mechanism of ciliary receptor targeting and functional role of Rab23 in promoting this process. DOI: http://dx.doi.org/10.7554/eLife.06996.001 PMID:26182404

  16. Multiple cilia suppress tumour formation.

    PubMed

    Eberhart, Charles

    2016-04-01

    Primary cilia are cellular structures that have important functions in development and disease. The suppression of multiciliate differentiation of choroid plexus precursors, and maintenance of a single primary cilium by Notch1, is now shown to be involved in choroid plexus tumour formation. PMID:27027488

  17. Cilia and coordination of signaling networks during heart development

    PubMed Central

    Koefoed, Karen; Veland, Iben Rønn; Pedersen, Lotte Bang; Larsen, Lars Allan; Christensen, Søren Tvorup

    2014-01-01

    Primary cilia are unique sensory organelles that coordinate a wide variety of different signaling pathways to control cellular processes during development and in tissue homeostasis. Defects in function or assembly of these antenna-like structures are therefore associated with a broad range of developmental disorders and diseases called ciliopathies. Recent studies have indicated a major role of different populations of cilia, including nodal and cardiac primary cilia, in coordinating heart development, and defects in these cilia are associated with congenital heart disease. Here, we present an overview of the role of nodal and cardiac primary cilia in heart development. PMID:24345806

  18. Cilia and cilia-associated proteins in cancer

    PubMed Central

    Seeger-Nukpezah, Tamina; Little, Joy L.; Serzhanova, Victoria; Golemis, Erica A.

    2013-01-01

    The primary cilium is a well-established target in the pathogenesis of numerous developmental and chronic disorders, and more recently is attracting interest as a structure relevant to cancer. Here we discuss mechanisms by which changes in cilia can contribute to the formation and growth of tumors. We emphasize the cancer-relevance of cilia-dependent signaling pathways and proteins including mTOR, VHL, TSC, WNT, Aurora-A, NEDD9, and Hedgehog, and highlight the emerging role of ciliary dysfunction in renal cell carcinoma, medulloblastoma, and breast cancer. PMID:24982684

  19. Non-essential role for cilia in coordinating precise alignment of lens fibres.

    PubMed

    Sugiyama, Yuki; Shelley, Elizabeth J; Yoder, Bradley K; Kozmik, Zbynek; May-Simera, Helen L; Beales, Philip L; Lovicu, Frank J; McAvoy, John W

    2016-02-01

    The primary cilium, a microtubule-based organelle found in most cells, is a centre for mechano-sensing fluid movement and cellular signalling, notably through the Hedgehog pathway. We recently found that each lens fibre cell has an apically situated primary cilium that is polarised to the side of the cell facing the anterior pole of the lens. The direction of polarity is similar in neighbouring cells so that in the global view, lens fibres exhibit planar cell polarity (PCP) along the equatorial-anterior polar axis. Ciliogenesis has been associated with the establishment of PCP, although the exact relationship between PCP and the role of cilia is still controversial. To test the hypothesis that the primary cilia have a role in coordinating the precise alignment/orientation of the fibre cells, IFT88, a key component of the intraflagellar transport (IFT) complex, was removed specifically from the lens at different developmental stages using several lens-specific Cre-expressing mouse lines (MLR10- and LR-Cre). Irrespective of which Cre-line was adopted, both demonstrated that in IFT88-depleted cells, the ciliary axoneme was absent or substantially shortened, confirming the disruption of primary cilia formation. However no obvious histological defects were detected even when IFT88 was removed from the lens placode as early as E9.5. Specifically, the lens fibres aligned/oriented towards the poles to form the characteristic Y-shaped sutures as normal. Consistent with this, in primary lens epithelial explants prepared from these conditional knockout mouse lenses, the basal bodies still showed polarised localisation at the apical surface of elongating cells upon FGF-induced fibre differentiation. We further investigated the lens phenotype in knockouts of Bardet-Biedl Syndrome (BBS) proteins 4 and 8, the components of the BBSome complex which modulate ciliary function. In these BBS4 and 8 knockout lenses, again we found the pattern of the anterior sutures formed by the

  20. The differential distribution of acetylated and detyrosinated alpha-tubulin in the microtubular cytoskeleton and primary cilia of hyaline cartilage chondrocytes

    PubMed Central

    POOLE, C. ANTHONY; ZHANG, ZI-JUN; ROSS, JACQUELINE M.

    2001-01-01

    The primary cilium is a ubiquitous cytoplasmic organelle of unknown function. Ultrastructural evidence of primary cilia in chondrocytes, and their colocalisation with the Golgi apparatus, has led to speculation that these structures are functionally linked. To investigate the relationship between these organelles, we examined the molecular anatomy of the microtubular cytoskeleton in the chondrocytes of chick embryo sterna. Thick cryosections were immunolabelled with antibodies directed against acetylated α-tubulin (C3B9), detyrosinated α-tubulin (ID5) and total α-tubulin (TAT), and imaged at high magnification using confocal laser scanning microscopy. Transmission electron microscopy confirmed the ultrastructure of the chondrocyte primary cilium and its structural relationship to the Golgi apparatus. Detyrosinated and acetylated α-tubulins were concentrated in the centrioles, centrosome and microtubule organising centre adjacent to the nucleus, with total α-tubulin distributed throughout the cytoplasm. ID5 stained the primary cilium at an incidence of 1 per cell, its colocalisation with C3B9 identifying the primary cilium as one of the most stable features of the microtubular cytoskeleton. Primary cilia varied from 1 to 4 μm in length, and 3 patterns of projection into the extracellular matrix were identified; (1) full extension and matrix contact, with minor undulations along the length; (2) partial extension and matrix contact, with a range of bending deflections; (3) cilium reclined against the cell surface with minimal matrix contact. Ultrastructural studies identified direct connections between extracellular collagen fibres and the proteins which decorate ciliary microtubules, suggesting a matrix–cilium–Golgi continuum in hyaline chondrocytes. These results strengthen the hypothesis that the primary cilium acts as a ‘cellular cybernetic probe' capable of transducing environmental information from the extracellular matrix, communicating this

  1. Primary cilia membrane assembly is initiated by Rab11 and transport protein particle II (TRAPPII) complex-dependent trafficking of Rabin8 to the centrosome

    PubMed Central

    Westlake, Christopher J.; Baye, Lisa M.; Nachury, Maxence V.; Wright, Kevin J.; Ervin, Karen E.; Phu, Lilian; Chalouni, Cecile; Beck, John S.; Kirkpatrick, Donald S.; Slusarski, Diane C.; Sheffield, Val C.; Scheller, Richard H.; Jackson, Peter K.

    2011-01-01

    Sensory and signaling pathways are exquisitely organized in primary cilia. Bardet-Biedl syndrome (BBS) patients have compromised cilia and signaling. BBS proteins form the BBSome, which binds Rabin8, a guanine nucleotide exchange factor (GEF) activating the Rab8 GTPase, required for ciliary assembly. We now describe serum-regulated upstream vesicular transport events leading to centrosomal Rab8 activation and ciliary membrane formation. Using live microscopy imaging, we show that upon serum withdrawal Rab8 is observed to assemble the ciliary membrane in ∼100 min. Rab8-dependent ciliary assembly is initiated by the relocalization of Rabin8 to Rab11-positive vesicles that are transported to the centrosome. After ciliogenesis, Rab8 ciliary transport is strongly reduced, and this reduction appears to be associated with decreased Rabin8 centrosomal accumulation. Rab11-GTP associates with the Rabin8 COOH-terminal region and is required for Rabin8 preciliary membrane trafficking to the centrosome and for ciliogenesis. Using zebrafish as a model organism, we show that Rabin8 and Rab11 are associated with the BBS pathway. Finally, using tandem affinity purification and mass spectrometry, we determined that the transport protein particle (TRAPP) II complex associates with the Rabin8 NH2-terminal domain and show that TRAPP II subunits colocalize with centrosomal Rabin8 and are required for Rabin8 preciliary targeting and ciliogenesis. PMID:21273506

  2. Sensory roles of neuronal cilia: cilia development, morphogenesis, and function in C. elegans.

    PubMed

    Bae, Young-Kyung; Barr, Maureen M

    2008-01-01

    In the free-living nematode Caenorhabditis elegans, cilia are found on the dendritic endings of sensory neurons. C. elegans cilia are classified as 'primary' or 'sensory' according to the '9+0' axonemal ultrastructure (nine doublet outer microtubules with no central microtubule pair) and lack of motility, characteristics of '9+2' cilia. The C. elegans ciliated nervous system allows the animal to perceive environmental stimuli and make appropriate developmental, physiological, and behavioral decisions. In vertebrates, the biological significance of primary cilia had been largely neglected. Recent findings have placed primary/sensory cilia in the center of cellular signaling and developmental processes. Studies using genetic model organisms such as C. elegans identified the link between ciliary dysfunction and human ciliopathies. Future studies in the worm will address important basic questions regarding ciliary development, morphogenesis, specialization, and signaling functions. PMID:18508635

  3. Polarisation of Light

    NASA Astrophysics Data System (ADS)

    Spottiswoode, William

    2015-01-01

    Preface; 1. Methods of polarisation; 2. Double refraction: polariscopes; 3. Chromatic polarisation: the wave theory; 4. Circular polarisation; 5. Circular polarisation by reflexion; 6. Phenomena produced by mechanical means: unannealed glass; 7. Atmospheric and other polarisation: the polar clock; 8. Rings and brushes produced by crystal plates; 9. Composition of colours by polarised light; Index.

  4. [Cilia and renal cysts].

    PubMed

    Paces-Fessy, Mélanie

    2014-11-01

    Advances in genomics, bioinformatics and the creation of model organisms have identified many genes associated with polycystic kidney diseases. Historically, these genes were not necessarily associated with ciliopathies, but it appeared that many connections can be made between the cystic kidney disease and function of the primary cilium. Indeed, the proteins encoded by these genes are localized to the cilium itself, to the basal body or are known to regulate the expression and localization of ciliary proteins. The goal of this article is to describe the multiple cellular processes that may lead to the development of renal cysts if they are deregulated. These include changes in proliferation rate, cell polarity or signaling pathways involved in embryonic kidney development. To highlight the role of the primary cilium in cystogenesis, I will discuss several studies investigating the function of ciliary genes and cilia in the kidneys of different model organisms. PMID:25388585

  5. Development and Distribution of Neuronal Cilia in Mouse Neocortex

    PubMed Central

    Arellano, Jon I.; Guadiana, Sarah M.; Breunig, Joshua J.; Rakic, Pasko; Sarkisian, Matthew R.

    2011-01-01

    Neuronal primary cilia are not generally recognized, but they are considered to extend from most, if not all, neurons in the neocortex. However, when and how cilia develop in neurons are not known. This study used immunohistochemistry for adenylyl cyclase III (ACIII), a marker of primary cilia, and electron microscopic analysis to describe the development and maturation of cilia in mouse neocortical neurons. Our results indicate that ciliogenesis is initiated in late fetal stages after neuroblast migration, when the mother centriole docks with the plasma membrane, becomes a basal body, and grows a cilia bud that we call a procilium. This procilium consists of a membranous protrusion extending from the basal body but lacking axonemal structure and remains undifferentiated until development of the axoneme and cilia elongation starts at about postnatal day 4. Neuronal cilia elongation and final cilia length depend on layer position, and the process extends for a long time, lasting 8–12 weeks. We show that, in addition to pyramidal neurons, inhibitory interneurons also grow cilia of comparable length, suggesting that cilia are indeed present in all neocortical neuron subtypes. Furthermore, the study of mice with defective ciliogenesis suggested that failed elongation of cilia is not essential for proper neuronal migration and laminar organization or establishment of neuronal polarity. Thus, the function of this organelle in neocortical neurons remains elusive. PMID:22020803

  6. Cilia, Wnt signaling, and the cytoskeleton.

    PubMed

    May-Simera, Helen L; Kelley, Matthew W

    2012-01-01

    Primary cilia have recently been highlighted as key regulators in development and disease. This review focuses on current work demonstrating the broad role of cilia-related proteins in developmental signaling systems. Of particular consideration is the importance of the basal body region, located at the base of the cilium, in its role as a focal point for many signaling pathways and as a microtubule organizing center. As the cilium is effectively a microtubular extension of the cytoskeleton, investigating connections between the cilium and the cytoskeleton provides greater insight into signaling and cell function. Of the many signaling pathways associated with primary cilia, the most extensively studied in association with the cytoskeleton and cytoskeletal rearrangements are both canonical and non-canonical Wnt pathways. One of the key concepts currently emerging is a possible additional role for the traditionally 'cilia-related' proteins in other aspects of cellular processes. In many cases, disruption of such processes manifests at the level of the cilium. While the involvement of cilia and cilia-related proteins in signaling pathways is currently being unraveled, there is a growing body of evidence to support the notion that ciliary proteins are required not only for regulation of Wnt signaling, but also as downstream effectors of Wnt signaling. This review summarizes recent advances in our understanding of the involvement of cilia and basal body proteins in Wnt signaling pathways. PMID:23351924

  7. KIF3A binds to β-arrestin for suppressing Wnt/β-catenin signalling independently of primary cilia in lung cancer.

    PubMed

    Kim, Minsuh; Suh, Young-Ah; Oh, Ju-Hee; Lee, Bo Ra; Kim, Joon; Jang, Se Jin

    2016-01-01

    Aberrant Wnt/β-catenin signalling is implicated in the progression of several human cancers, including non-small cell lung cancer (NSCLC). However, mutations in Wnt/β-catenin pathway components are uncommon in NSCLC, and their epigenetic control remains unclear. Here, we show that KIF3A, a member of the kinesin-2 family, plays a role in suppressing Wnt/β-catenin signalling in NSCLC cells. KIF3A knockdown increases both β-catenin levels and transcriptional activity with concomitant promotion of malignant potential, such as increased proliferation and migration and upregulation of stemness markers. Because KIF3A binds β-arrestin, KIF3A depletion allows β-arrestin to form a complex with DVL2 and axin, stabilizing β-catenin. Although primary cilia, whose biogenesis requires KIF3A, are thought to restrain the Wnt response, pharmacological inhibition of ciliogenesis failed to increase β-catenin activity in NSCLC cells. A correlation between KIF3A loss and a poorer NSCLC prognosis as well as β-catenin and cyclin D1 upregulation further suggests that KIF3A suppresses Wnt/β-catenin signalling and tumourigenesis in NSCLC. PMID:27596264

  8. KIF3A binds to β-arrestin for suppressing Wnt/β-catenin signalling independently of primary cilia in lung cancer

    PubMed Central

    Kim, Minsuh; Suh, Young-Ah; Oh, Ju-Hee; Lee, Bo Ra; Kim, Joon; Jang, Se Jin

    2016-01-01

    Aberrant Wnt/β-catenin signalling is implicated in the progression of several human cancers, including non-small cell lung cancer (NSCLC). However, mutations in Wnt/β-catenin pathway components are uncommon in NSCLC, and their epigenetic control remains unclear. Here, we show that KIF3A, a member of the kinesin-2 family, plays a role in suppressing Wnt/β-catenin signalling in NSCLC cells. KIF3A knockdown increases both β-catenin levels and transcriptional activity with concomitant promotion of malignant potential, such as increased proliferation and migration and upregulation of stemness markers. Because KIF3A binds β-arrestin, KIF3A depletion allows β-arrestin to form a complex with DVL2 and axin, stabilizing β-catenin. Although primary cilia, whose biogenesis requires KIF3A, are thought to restrain the Wnt response, pharmacological inhibition of ciliogenesis failed to increase β-catenin activity in NSCLC cells. A correlation between KIF3A loss and a poorer NSCLC prognosis as well as β-catenin and cyclin D1 upregulation further suggests that KIF3A suppresses Wnt/β-catenin signalling and tumourigenesis in NSCLC. PMID:27596264

  9. Mutations in CEP78 Cause Cone-Rod Dystrophy and Hearing Loss Associated with Primary-Cilia Defects.

    PubMed

    Nikopoulos, Konstantinos; Farinelli, Pietro; Giangreco, Basilio; Tsika, Chrysanthi; Royer-Bertrand, Beryl; Mbefo, Martial K; Bedoni, Nicola; Kjellström, Ulrika; El Zaoui, Ikram; Di Gioia, Silvio Alessandro; Balzano, Sara; Cisarova, Katarina; Messina, Andrea; Decembrini, Sarah; Plainis, Sotiris; Blazaki, Styliani V; Khan, Muhammad Imran; Micheal, Shazia; Boldt, Karsten; Ueffing, Marius; Moulin, Alexandre P; Cremers, Frans P M; Roepman, Ronald; Arsenijevic, Yvan; Tsilimbaris, Miltiadis K; Andréasson, Sten; Rivolta, Carlo

    2016-09-01

    Cone-rod degeneration (CRD) belongs to the disease spectrum of retinal degenerations, a group of hereditary disorders characterized by an extreme clinical and genetic heterogeneity. It mainly differentiates from other retinal dystrophies, and in particular from the more frequent disease retinitis pigmentosa, because cone photoreceptors degenerate at a higher rate than rod photoreceptors, causing severe deficiency of central vision. After exome analysis of a cohort of individuals with CRD, we identified biallelic mutations in the orphan gene CEP78 in three subjects from two families: one from Greece and another from Sweden. The Greek subject, from the island of Crete, was homozygous for the c.499+1G>T (IVS3+1G>T) mutation in intron 3. The Swedish subjects, two siblings, were compound heterozygotes for the nearby mutation c.499+5G>A (IVS3+5G>A) and for the frameshift-causing variant c.633delC (p.Trp212Glyfs(∗)18). In addition to CRD, these three individuals had hearing loss or hearing deficit. Immunostaining highlighted the presence of CEP78 in the inner segments of retinal photoreceptors, predominantly of cones, and at the base of the primary cilium of fibroblasts. Interaction studies also showed that CEP78 binds to FAM161A, another ciliary protein associated with retinal degeneration. Finally, analysis of skin fibroblasts derived from affected individuals revealed abnormal ciliary morphology, as compared to that of control cells. Altogether, our data strongly suggest that mutations in CEP78 cause a previously undescribed clinical entity of a ciliary nature characterized by blindness and deafness but clearly distinct from Usher syndrome, a condition for which visual impairment is due to retinitis pigmentosa. PMID:27588451

  10. Cilia and Polycystic Kidney Disease, Kith and Kin

    PubMed Central

    Huang, Liwei; Lipschutz, Joshua H.

    2015-01-01

    In the past decade, cilia have been found to play important roles in renal cystogenesis. Many genes, such as PKD1 and PKD2 which, when mutated, cause autosomal dominant polycystic kidney disease (ADPKD), have been found to localize to primary cilia. The cilium functions as a sensor to transmit extracellular signals into the cell. Abnormal cilia structure and function are associated with the development of polyscystic kidney disease (PKD). Cilia assembly includes centriole migration to the apical surface of the cell, ciliary vesicle docking and fusion with the cell membrane at the intended site of cilium outgrowth, and microtubule growth from the basal body. This review summarizes the most recent advances in cilia and PKD research, with special emphasis on the mechanisms of cytoplasmic and intraciliary protein transport during ciliogenesis. PMID:24898006

  11. Genetic Ablation of Type III Adenylyl Cyclase Exerts Region-Specific Effects on Cilia Architecture in the Mouse Nose

    PubMed Central

    Challis, Rosemary C.; Tian, Huikai; Yin, Wenbin; Ma, Minghong

    2016-01-01

    We recently reported that olfactory sensory neurons in the dorsal zone of the mouse olfactory epithelium exhibit drastic location-dependent differences in cilia length. Furthermore, genetic ablation of type III adenylyl cyclase (ACIII), a key olfactory signaling protein and ubiquitous marker for primary cilia, disrupts the cilia length pattern and results in considerably shorter cilia, independent of odor-induced activity. Given the significant impact of ACIII on cilia length in the dorsal zone, we sought to further investigate the relationship between cilia length and ACIII level in various regions throughout the mouse olfactory epithelium. We employed whole-mount immunohistochemical staining to examine olfactory cilia morphology in phosphodiesterase (PDE) 1C-/-;PDE4A-/- (simplified as PDEs-/- hereafter) and ACIII-/- mice in which ACIII levels are reduced and ablated, respectively. As expected, PDEs-/- animals exhibit dramatically shorter cilia in the dorsal zone (i.e., where the cilia pattern is found), similar to our previous observation in ACIII-/- mice. Remarkably, in a region not included in our previous study, ACIII-/- animals (but not PDEs-/- mice) have dramatically elongated, comet-shaped cilia, as opposed to characteristic star-shaped olfactory cilia. Here, we reveal that genetic ablation of ACIII has drastic, location-dependent effects on cilia architecture in the mouse nose. These results add a new dimension to our current understanding of olfactory cilia structure and regional organization of the olfactory epithelium. Together, these findings have significant implications for both cilia and sensory biology. PMID:26942602

  12. Autophagy and regulation of cilia function and assembly

    PubMed Central

    Orhon, I; Dupont, N; Pampliega, O; Cuervo, A M; Codogno, P

    2015-01-01

    Motile and primary cilia (PC) are microtubule-based structures located at the cell surface of many cell types. Cilia govern cellular functions ranging from motility to integration of mechanical and chemical signaling from the environment. Recent studies highlight the interplay between cilia and autophagy, a conserved cellular process responsible for intracellular degradation. Signaling from the PC recruits the autophagic machinery to trigger autophagosome formation. Conversely, autophagy regulates ciliogenesis by controlling the levels of ciliary proteins. The cross talk between autophagy and ciliated structures is a novel aspect of cell biology with major implications in development, physiology and human pathologies related to defects in cilium function. PMID:25361082

  13. In vivo investigation of cilia structure and function using Xenopus

    PubMed Central

    Brooks, Eric R.; Wallingford, John B.

    2015-01-01

    Cilia are key organelles in development and homeostasis. The ever-expanding complement of cilia associated proteins necessitates rapid and tractable models for in vivo functional investigation. Xenopus laevis provides an attractive model for such studies, having multiple ciliated populations, including primary and multiciliated tissues. The rapid external development of Xenopus and the large cells make it an especially excellent platform for imaging studies. Here we present embryological and cell-biological methods for the investigation of cilia structure and function in Xenopus laevis, with a focus on quantitative live and fixed imaging. PMID:25837389

  14. Cilia/Ift protein and motor-related bone diseases and mouse models

    PubMed Central

    Yuan, Xue; Yang, Shuying

    2015-01-01

    Primary cilia are essential cellular organelles projecting from the cell surface to sense and transduce developmental signaling. They are tiny but have complicated structures containing microtubule (MT)-based internal structures (the axoneme) and mother centriole formed basal body. Intraflagellar transport (Ift) operated by Ift proteins and motors are indispensable for cilia formation and function. Mutations in Ift proteins or Ift motors cause various human diseases, some of which have severe bone defects. Over the last few decades, major advances have occurred in understanding the roles of these proteins and cilia in bone development and remodeling by examining cilia/Ift protein-related human diseases and establishing mouse transgenic models. In this review, we describe current advances in the understanding of the cilia/Ift structure and function. We further summarize cilia/Ift-related human diseases and current mouse models with an emphasis on bone-related phenotypes, cilia morphology, and signaling pathways. PMID:25553465

  15. Reduction of the immunostainable length of the hippocampal dentate granule cells' primary cilia in 3xAD-transgenic mice producing human A{beta}{sub 1-42} and tau

    SciTech Connect

    Chakravarthy, Balu; Gaudet, Chantal; Menard, Michel; Brown, Leslie; Atkinson, Trevor; LaFerla, Frank M.; Ito, Shingo; Armato, Ubaldo; Dal Pra, Ilaria; Whitfield, James

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer A{beta} and tau-induced neurofibrillary tangles play a key role in Alzheimer's disease. Black-Right-Pointing-Pointer A{beta}{sub 1-42} and mutant tau protein together reduce the primary cilium length. Black-Right-Pointing-Pointer This shortening likely reduces cilium-dependent neurogenesis and memory function. Black-Right-Pointing-Pointer This provides a model of an A{beta}/tau targeting of a neuronal signaling organelle. -- Abstract: The hippocampal dentate gyrus is one of the two sites of continuous neurogenesis in adult rodents and humans. Virtually all dentate granule cells have a single immobile cilium with a microtubule spine or axoneme covered with a specialized cell membrane loaded with receptors such as the somatostatin receptor 3 (SSTR3), and the p75 neurotrophin receptor (p75{sup NTR}). The signals from these receptors have been reported to stimulate neuroprogenitor proliferation and the post-mitotic maturation of newborn granule cells into functioning granule cells. We have found that in 6-24-months-old triple transgenic Alzheimer's disease model mice (3xTg-AD) producing both A{beta}{sub 1-42} and the mutant human tau protein tau{sub P301L,} the dentate granule cells still had immunostainable SSTR3- and p75{sup NTR}-bearing cilia but they were only half the length of the immunostained cilia in the corresponding wild-type mice. However, the immunostainable length of the granule cell cilia was not reduced either in 2xTg-AD mice accumulating large amounts of A{beta}{sub 1-42} or in mice accumulating only a mutant human tau protein. Thus it appears that a combination of A{beta}{sub 1-42} and tau protein accumulation affects the levels of functionally important receptors in 3xTg-AD mice. These observations raise the important possibility that structural and functional changes in granule cell cilia might have a role in AD.

  16. Cyclic GMP and Cilia Motility

    PubMed Central

    Wyatt, Todd A.

    2015-01-01

    Motile cilia of the lungs respond to environmental challenges by increasing their ciliary beat frequency in order to enhance mucociliary clearance as a fundamental tenant of innate defense. One important second messenger in transducing the regulable nature of motile cilia is cyclic guanosine 3′,5′-monophosphate (cGMP). In this review, the history of cGMP action is presented and a survey of the existing data addressing cGMP action in ciliary motility is presented. Nitric oxide (NO)-mediated regulation of cGMP in ciliated cells is presented in the context of alcohol-induced cilia function and dysfunction. PMID:26264028

  17. Olfactory Cilia: Linking Sensory Cilia Function and Human Disease

    PubMed Central

    Jenkins, Paul M.; McEwen, Dyke P.

    2009-01-01

    The olfactory system gives us an awareness of our immediate environment by allowing us to detect airborne stimuli. The components necessary for detection of these odorants are compartmentalized in the cilia of olfactory sensory neurons. Cilia are microtubule-based organelles, which can be found projecting from the surface of almost any mammalian cell, and are critical for proper olfactory function. Mislocalization of ciliary proteins and/or the loss of cilia cause impaired olfactory function, which is now recognized as a clinical manifestation of a broad class of human diseases, termed ciliopathies. Future work investigating the mechanisms of olfactory cilia function will provide us important new information regarding the pathogenesis of human sensory perception diseases. PMID:19406873

  18. Antennas of organ morphogenesis: the roles of cilia in vertebrate kidney development.

    PubMed

    Marra, Amanda N; Li, Yue; Wingert, Rebecca A

    2016-09-01

    Cilia arose early during eukaryotic evolution, and their structural components are highly conserved from the simplest protists to complex metazoan species. In recent years, the role of cilia in the ontogeny of vertebrate organs has received increasing attention due to a staggering correlation between human disease and dysfunctional cilia. In particular, the presence of cilia in both the developing and mature kidney has become a deep area of research due to ciliopathies common to the kidney, such as polycystic kidney disease (PKD). Interestingly, mutations in genes encoding proteins that localize to the cilia cause similar cystic phenotypes in kidneys of various vertebrates, suggesting an essential role for cilia in kidney organogenesis and homeostasis as well. Importantly, the genes so far identified in kidney disease have conserved functions across species, whose kidneys include both primary and motile cilia. Here, we aim to provide a comprehensive description of cilia and their role in kidney development, as well as highlight the usefulness of the zebrafish embryonic kidney as a model to further understand the function of cilia in kidney health. PMID:27389733

  19. Evaluating efficiency and robustness in cilia design

    NASA Astrophysics Data System (ADS)

    Guo, Hanliang; Kanso, Eva

    2016-03-01

    Motile cilia are used by many eukaryotic cells to transport flow. Cilia-driven flows are important to many physiological functions, yet a deep understanding of the interplay between the mechanical structure of cilia and their physiological functions in healthy and diseased conditions remains elusive. To develop such an understanding, one needs a quantitative framework to assess cilia performance and robustness when subject to perturbations in the cilia apparatus. Here we link cilia design (beating patterns) to function (flow transport) in the context of experimentally and theoretically derived cilia models. We particularly examine the optimality and robustness of cilia design. Optimality refers to efficiency of flow transport, while robustness is defined as low sensitivity to variations in the design parameters. We find that suboptimal designs can be more robust than optimal ones. That is, designing for the most efficient cilium does not guarantee robustness. These findings have significant implications on the understanding of cilia design in artificial and biological systems.

  20. Evaluating efficiency and robustness in cilia design.

    PubMed

    Guo, Hanliang; Kanso, Eva

    2016-03-01

    Motile cilia are used by many eukaryotic cells to transport flow. Cilia-driven flows are important to many physiological functions, yet a deep understanding of the interplay between the mechanical structure of cilia and their physiological functions in healthy and diseased conditions remains elusive. To develop such an understanding, one needs a quantitative framework to assess cilia performance and robustness when subject to perturbations in the cilia apparatus. Here we link cilia design (beating patterns) to function (flow transport) in the context of experimentally and theoretically derived cilia models. We particularly examine the optimality and robustness of cilia design. Optimality refers to efficiency of flow transport, while robustness is defined as low sensitivity to variations in the design parameters. We find that suboptimal designs can be more robust than optimal ones. That is, designing for the most efficient cilium does not guarantee robustness. These findings have significant implications on the understanding of cilia design in artificial and biological systems. PMID:27078459

  1. Type 3 Adenylyl Cyclase and Somatostatin Receptor 3 Expression Persists in Aged Rat Neocortical and Hippocampal Neuronal Cilia

    PubMed Central

    Guadiana, Sarah M.; Parker, Alexander K.; Filho, Gileno F.; Sequeira, Ashton; Semple-Rowland, Susan; Shaw, Gerry; Mandel, Ronald J.; Foster, Thomas C.; Kumar, Ashok; Sarkisian, Matthew R.

    2016-01-01

    The primary cilia of forebrain neurons assemble around birth and become enriched with neuromodulatory receptors. Our understanding of the permanence of these structures and their associated signaling pathways in the aging brain is poor, but they are worthy of investigation because disruptions in neuronal cilia signaling have been implicated in changes in learning and memory, depression-like symptoms, and sleep anomalies. Here, we asked whether neurons in aged forebrain retain primary cilia and whether the staining characteristics of aged cilia for type 3 adenylyl cyclase (ACIII), somatostatin receptor 3 (SSTR3), and pericentrin resemble those of cilia in younger forebrain. To test this, we analyzed immunostained sections of forebrain tissues taken from young and aged male Fischer 344 (F344) and F344 × Brown Norway (F344 × BN) rats. Analyses of ACIII and SSTR3 in young and aged cortices of both strains of rats revealed that the staining patterns in the neocortex and hippocampus were comparable. Virtually every NeuN positive cell examined possessed an ACIII positive cilium. The lengths of ACIII positive cilia in neocortex were similar between young and aged for both strains, whereas in F344 × BN hippocampus, the cilia lengths increased with age in CA1 and CA3, but not in dentate gyrus (DG). Additionally, the percentages of ACIII positive cilia that were also SSTR3 positive did not differ between young and aged tissues in either strain. We also found that pericentrin, a protein that localizes to the basal bodies of neuronal cilia and functions in primary cilia assembly, persisted in aged cortical neurons of both rat strains. Collectively, our data show that neurons in aged rat forebrain possess primary cilia and that these cilia, like those present in younger brain, continue to localize ACIII, SSTR3, and pericentrin. Further studies will be required to determine if the function and signaling pathways regulated by cilia are similar in aged compared to young brain

  2. Polarised light sheet tomography.

    PubMed

    Reidt, Sascha L; O'Brien, Daniel J; Wood, Kenneth; MacDonald, Michael P

    2016-05-16

    The various benefits of light sheet microscopy have made it a widely used modality for capturing three-dimensional images. It is mostly used for fluorescence imaging, but recently another technique called light sheet tomography solely relying on scattering was presented. The method was successfully applied to imaging of plant roots in transparent soil, but is limited when it comes to more turbid samples. This study presents a polarised light sheet tomography system and its advantages when imaging in highly scattering turbid media. The experimental configuration is guided by Monte Carlo radiation transfer methods, which model the propagation of a polarised light sheet in the sample. Images of both reflecting and absorbing phantoms in a complex collagenous matrix were acquired, and the results for different polarisation configurations are compared. Focus scanning methods were then used to reduce noise and produce three-dimensional reconstructions of absorbing targets. PMID:27409945

  3. Polarisation properties of pulsars at optical wavelengths

    NASA Astrophysics Data System (ADS)

    Mignani, Roberto; Marelli, Martino; Shearer, Andrew; Slowikowska, Agnieszka

    2016-07-01

    Polarisation measurements of pulsars offer unique insights into their highly-magnetised relativistic environments and represent a primary test for neutron star magnetosphere models and radiation emission mechanisms. Besides the radio band, optical observations have been, so far, best suited to these goals, with polarisation measurements in the X-rays becoming possible in the near future thanks to missions, such as XIPE and IXPE. In this talk, we review the status of the optical polarisation measurements of pulsars and we foresee possible synergies between X-ray polarimetry observations of selected pulsars with, e.g XIPE and IXPE, and optical observations with the next generation of extremely large telescope, such as the E-ELT.

  4. Tau polarisation at LEP

    NASA Astrophysics Data System (ADS)

    Alemany, Ricard

    1999-04-01

    The measurements of the tau polarisation at LEP I are reviewed. Special emphasis is given to the new preliminary results presented at this conference. The ALEPH collaboration has studied the polarisation as a function of the polar angle using a new method based on the tau direction reconstruction and fully exploiting the angular correlations. A second traditional approach, based on the single tau decays has been also developed. The DELPHI collaboration has also studied the full data sample using an individual tau decay method and an inclusive hadronic selection. The results from the four experiments are presented with discussion of the compatibility among the methods and experiments.

  5. Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease.

    PubMed

    Lewis, Wesley R; Malarkey, Erik B; Tritschler, Douglas; Bower, Raqual; Pasek, Raymond C; Porath, Jonathan D; Birket, Susan E; Saunier, Sophie; Antignac, Corinne; Knowles, Michael R; Leigh, Margaret W; Zariwala, Maimoona A; Challa, Anil K; Kesterson, Robert A; Rowe, Steven M; Drummond, Iain A; Parant, John M; Hildebrandt, Friedhelm; Porter, Mary E; Yoder, Bradley K; Berbari, Nicolas F

    2016-07-01

    Ciliopathies are genetic disorders arising from dysfunction of microtubule-based cellular appendages called cilia. Different cilia types possess distinct stereotypic microtubule doublet arrangements with non-motile or 'primary' cilia having a 9+0 and motile cilia have a 9+2 array of microtubule doublets. Primary cilia are critical sensory and signaling centers needed for normal mammalian development. Defects in their structure/function result in a spectrum of clinical and developmental pathologies including abnormal neural tube and limb patterning. Altered patterning phenotypes in the limb and neural tube are due to perturbations in the hedgehog (Hh) signaling pathway. Motile cilia are important in fluid movement and defects in motility result in chronic respiratory infections, altered left-right asymmetry, and infertility. These features are the hallmarks of Primary Ciliary Dyskinesia (PCD, OMIM 244400). While mutations in several genes are associated with PCD in patients and animal models, the genetic lesion in many cases is unknown. We assessed the in vivo functions of Growth Arrest Specific 8 (GAS8). GAS8 shares strong sequence similarity with the Chlamydomonas Nexin-Dynein Regulatory Complex (NDRC) protein 4 (DRC4) where it is needed for proper flagella motility. In mammalian cells, the GAS8 protein localizes not only to the microtubule axoneme of motile cilia, but also to the base of non-motile cilia. Gas8 was recently implicated in the Hh signaling pathway as a regulator of Smoothened trafficking into the cilium. Here, we generate the first mouse with a Gas8 mutation and show that it causes severe PCD phenotypes; however, there were no overt Hh pathway phenotypes. In addition, we identified two human patients with missense variants in Gas8. Rescue experiments in Chlamydomonas revealed a subtle defect in swim velocity compared to controls. Further experiments using CRISPR/Cas9 homology driven repair (HDR) to generate one of these human missense variants in

  6. Emergence of metachronal waves in cilia arrays

    PubMed Central

    Elgeti, Jens; Gompper, Gerhard

    2013-01-01

    Propulsion by cilia is a fascinating and universal mechanism in biological organisms to generate fluid motion on the cellular level. Cilia are hair-like organelles, which are found in many different tissues and many uni- and multicellular organisms. Assembled in large fields, cilia beat neither randomly nor completely synchronously—instead they display a striking self-organization in the form of metachronal waves (MCWs). It was speculated early on that hydrodynamic interactions provide the physical mechanism for the synchronization of cilia motion. Theory and simulations of physical model systems, ranging from arrays of highly simplified actuated particles to a few cilia or cilia chains, support this hypothesis. The main questions are how the individual cilia interact with the flow field generated by their neighbors and synchronize their beats for the metachronal wave to emerge and how the properties of the metachronal wave are determined by the geometrical arrangement of the cilia, like cilia spacing and beat direction. Here, we address these issues by large-scale computer simulations of a mesoscopic model of 2D cilia arrays in a 3D fluid medium. We show that hydrodynamic interactions are indeed sufficient to explain the self-organization of MCWs and study beat patterns, stability, energy expenditure, and transport properties. We find that the MCW can increase propulsion velocity more than 3-fold and efficiency almost 10-fold—compared with cilia all beating in phase. This can be a vital advantage for ciliated organisms and may be interesting to guide biological experiments as well as the design of efficient microfluidic devices and artificial microswimmers. PMID:23487771

  7. Emergence of metachronal waves in cilia arrays.

    PubMed

    Elgeti, Jens; Gompper, Gerhard

    2013-03-19

    Propulsion by cilia is a fascinating and universal mechanism in biological organisms to generate fluid motion on the cellular level. Cilia are hair-like organelles, which are found in many different tissues and many uni- and multicellular organisms. Assembled in large fields, cilia beat neither randomly nor completely synchronously--instead they display a striking self-organization in the form of metachronal waves (MCWs). It was speculated early on that hydrodynamic interactions provide the physical mechanism for the synchronization of cilia motion. Theory and simulations of physical model systems, ranging from arrays of highly simplified actuated particles to a few cilia or cilia chains, support this hypothesis. The main questions are how the individual cilia interact with the flow field generated by their neighbors and synchronize their beats for the metachronal wave to emerge and how the properties of the metachronal wave are determined by the geometrical arrangement of the cilia, like cilia spacing and beat direction. Here, we address these issues by large-scale computer simulations of a mesoscopic model of 2D cilia arrays in a 3D fluid medium. We show that hydrodynamic interactions are indeed sufficient to explain the self-organization of MCWs and study beat patterns, stability, energy expenditure, and transport properties. We find that the MCW can increase propulsion velocity more than 3-fold and efficiency almost 10-fold--compared with cilia all beating in phase. This can be a vital advantage for ciliated organisms and may be interesting to guide biological experiments as well as the design of efficient microfluidic devices and artificial microswimmers. PMID:23487771

  8. Magnetically Actuated Cilia for Microfluidic Manipulation

    NASA Astrophysics Data System (ADS)

    Hanasoge, Srinivas; Owen, Drew; Ballard, Matt; Hesketh, Peter J.; Alexeev, Alexander; Woodruff School of Mechanical Engineering Collaboration; Petit InstituteBioengineering; Biosciences Collaboration

    2015-11-01

    We demonstrate magnetic micro-cilia based microfluidic mixing and capture techniques. For this, we use a simple and easy to fabricate high aspect ratio cilia, which are actuated magnetically. These micro-features are fabricated by evaporating NiFe alloy at room temperature, on to patterned photoresist. The evaporated alloy curls upwards when the seed layer is removed to release the cilia, thus making a free standing `C' shaped magnetic microstructure. This is actuated using an external electromagnet or a rotating magnet. The artificial cilia can be actuated upto 20Hz. We demonstrate the active mixing these cilia can produce in the microchannel. Also, we demonstrate the capture of target species in a sample using these fast oscillating cilia. The surface of the cilia is functionalized by streptavidin which binds to biotin labelled fluorescent microspheres and mimic the capture of bacteria. We show very high capture efficiencies by using these methods. These simple to fabricate micro cilia can easily be incorporated into many microfluidic systems which require high mixing and capture efficiencies.

  9. Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease

    PubMed Central

    Lewis, Wesley R.; Malarkey, Erik B.; Tritschler, Douglas; Bower, Raqual; Pasek, Raymond C.; Porath, Jonathan D.; Birket, Susan E.; Saunier, Sophie; Antignac, Corinne; Leigh, Margaret W.; Zariwala, Maimoona A.; Drummond, Iain A.; Parant, John M.; Hildebrandt, Friedhelm; Yoder, Bradley K.

    2016-01-01

    Ciliopathies are genetic disorders arising from dysfunction of microtubule-based cellular appendages called cilia. Different cilia types possess distinct stereotypic microtubule doublet arrangements with non-motile or ‘primary’ cilia having a 9+0 and motile cilia have a 9+2 array of microtubule doublets. Primary cilia are critical sensory and signaling centers needed for normal mammalian development. Defects in their structure/function result in a spectrum of clinical and developmental pathologies including abnormal neural tube and limb patterning. Altered patterning phenotypes in the limb and neural tube are due to perturbations in the hedgehog (Hh) signaling pathway. Motile cilia are important in fluid movement and defects in motility result in chronic respiratory infections, altered left-right asymmetry, and infertility. These features are the hallmarks of Primary Ciliary Dyskinesia (PCD, OMIM 244400). While mutations in several genes are associated with PCD in patients and animal models, the genetic lesion in many cases is unknown. We assessed the in vivo functions of Growth Arrest Specific 8 (GAS8). GAS8 shares strong sequence similarity with the Chlamydomonas Nexin-Dynein Regulatory Complex (NDRC) protein 4 (DRC4) where it is needed for proper flagella motility. In mammalian cells, the GAS8 protein localizes not only to the microtubule axoneme of motile cilia, but also to the base of non-motile cilia. Gas8 was recently implicated in the Hh signaling pathway as a regulator of Smoothened trafficking into the cilium. Here, we generate the first mouse with a Gas8 mutation and show that it causes severe PCD phenotypes; however, there were no overt Hh pathway phenotypes. In addition, we identified two human patients with missense variants in Gas8. Rescue experiments in Chlamydomonas revealed a subtle defect in swim velocity compared to controls. Further experiments using CRISPR/Cas9 homology driven repair (HDR) to generate one of these human missense variants

  10. Swimming like algae: biomimetic soft artificial cilia.

    PubMed

    Sareh, Sina; Rossiter, Jonathan; Conn, Andrew; Drescher, Knut; Goldstein, Raymond

    2013-01-01

    Cilia are used effectively in a wide variety of biological systems from fluid transport to thrust generation. Here, we present the design and implementation of artificial cilia, based on a biomimetic planar actuator using soft-smart materials. This actuator is modelled on the cilia movement of the alga Volvox, and represents the cilium as a piecewise constant-curvature robotic actuator that enables the subsequent direct translation of natural articulation into a multi-segment ionic polymer metal composite actuator. It is demonstrated how the combination of optimal segmentation pattern and biologically derived per-segment driving signals reproduce natural ciliary motion. The amenability of the artificial cilia to scaling is also demonstrated through the comparison of the Reynolds number achieved with that of natural cilia. PMID:23097503

  11. Swimming like algae: biomimetic soft artificial cilia

    PubMed Central

    Sareh, Sina; Rossiter, Jonathan; Conn, Andrew; Drescher, Knut; Goldstein, Raymond E.

    2013-01-01

    Cilia are used effectively in a wide variety of biological systems from fluid transport to thrust generation. Here, we present the design and implementation of artificial cilia, based on a biomimetic planar actuator using soft-smart materials. This actuator is modelled on the cilia movement of the alga Volvox, and represents the cilium as a piecewise constant-curvature robotic actuator that enables the subsequent direct translation of natural articulation into a multi-segment ionic polymer metal composite actuator. It is demonstrated how the combination of optimal segmentation pattern and biologically derived per-segment driving signals reproduce natural ciliary motion. The amenability of the artificial cilia to scaling is also demonstrated through the comparison of the Reynolds number achieved with that of natural cilia. PMID:23097503

  12. Microfluidic manipulation with artificial/bioinspired cilia.

    PubMed

    den Toonder, Jaap M J; Onck, Patrick R

    2013-02-01

    A recent development, inspired by nature, is the use of 'artificial cilia' to create pumping and/or mixing in microfluidic devices. Cilia are small hairs that can be found in biology and are used for (fluid) actuation and sensing. Microscopic actuators resembling cilia, actuated to move under the influence of various stimuli such as electrostatic field, magnetic field, and even light, have been developed by a number of groups and shown to be capable of generating flow and mixing in microfluidic environments. The research on artificial cilia started about a decade ago and is rapidly expanding. In addition to being relevant for potential application in lab-on-a-chip devices, the work on artificial cilia forms a beautiful example of how a biological system can form the successful basis for both scientific research and technological applications. In this review, we will give an overview of the most important approaches in this exciting field. PMID:23245658

  13. Loss of Bardet–Biedl syndrome proteins alters the morphology and function of motile cilia in airway epithelia

    PubMed Central

    Shah, Alok S.; Farmen, Sara L.; Moninger, Thomas O.; Businga, Thomas R.; Andrews, Michael P.; Bugge, Kevin; Searby, Charles C.; Nishimura, Darryl; Brogden, Kim A.; Kline, Joel N.; Sheffield, Val C.; Welsh, Michael J.

    2008-01-01

    Mutations in a group of genes that contribute to ciliary function cause Bardet–Biedl syndrome (BBS). Most studies of BBS have focused on primary, sensory cilia. Here, we asked whether loss of BBS proteins would also affect motile cilia lining the respiratory tract. We found that BBS genes were expressed in human airway epithelia, and BBS2 and BBS4 localized to cellular structures associated with motile cilia. Although BBS proteins were not required for ciliogenesis, their loss caused structural defects in a fraction of cilia covering mouse airway epithelia. The most common abnormality was bulges filled with vesicles near the tips of cilia. We discovered this same misshapen appearance in airway cilia from Bbs1, Bbs2, Bbs4, and Bbs6 mutant mice. The structural abnormalities were accompanied by functional defects; ciliary beat frequency was reduced in Bbs mutant mice. Previous reports suggested BBS might increase the incidence of asthma. However, compared with wild-type controls, neither airway hyperresponsiveness nor inflammation increased in Bbs2−/− or Bbs4−/− mice immunized with ovalbumin. Instead, these animals were partially protected from airway hyperresponsiveness. These results emphasize the role of BBS proteins in both the structure and function of motile cilia. They also invite additional scrutiny of motile cilia dysfunction in patients with this disease. PMID:18299575

  14. Polarised neutron scattering from dynamic polarised targets in biology

    NASA Astrophysics Data System (ADS)

    Knop, W.; Hirai, M.; Olah, G.; Meerwinck, W.; Schink, H.-J.; Stuhrman, H. B.; Wagner, R.; Wenkow-EsSouni, M.; Zhao, J.; Schärpf, O.; Crichton, R. R.; Krumpolc, M.; Nierhaus, K. H.; Niinikoski, T. O.; Rijllart, A.

    1991-10-01

    The contrast giving rise to neutron small-angle scattering can be enhanced considerably by polarisation of the hydrogen nuclei [J. des Coizeaux and G. Jannink, Les Polymères en Solution, Les Editions de Physique, F-91944 Les Ulis, France (1987)]. Using polarised neutrons the scattering from protonated labels in a deuterated matrix will increase by an order of magnitude. This is the basis of nuclear spin contrast variation, a method which is of particular interest for the in situ structure determination of macromolecular components. A new polarised target for neutron scattering has been designed by CERN and tested successfully at FRG-1 of the GKSS research centre. For the purpose of thermal-neutron scattering the frozen solutions of biomolecules are immersed in liquid helium 4, which is thermally coupled to the cooling mixture of helium 3/helium 4 of the dilution refrigerator. The nuclear spins are aligned with respect to the external magnetic field-parallel or antiparallel-by dynamic nuclear polarisation (DNP). The gain in neutron scattering compared to earlier experiments using direct cooling of the sample by helium 3 is a factor of 30. Another factor of 30 arises from the installation of the cold source and the beryllium reflector in FRG-1 [W. Knop et al., J. Appl. Cryst. 22 (1989) 352]. Pure nuclear spin targets are produced from dynamic polarised targets by selective depolarisation. In biological material only the hydrogen isotopes contribute significantly to polarised neutron scattering. Thus, saturation of the proton NMR yields a deuteron target, provided the target material has been enriched by the latter isotope. A proton target is obtained from the dynamic polarised target by saturation of deuteron NMR. This leads to six additional scattering functions reflecting the proton and deuteron spin densities and the correlations between the polarised isotopes. Polarised neutron scattering from nuclear spin targets of apoferritin and various derivatives of the

  15. Centrosomal protein CP110 controls maturation of the mother centriole during cilia biogenesis

    PubMed Central

    Yadav, Sharda Prasad; Sharma, Neel Kamal; Liu, Chunqiao; Dong, Lijin; Li, Tiansen; Swaroop, Anand

    2016-01-01

    ABSTRACT Defects in cilia centrosomal genes cause pleiotropic clinical phenotypes, collectively called ciliopathies. Cilia biogenesis is initiated by the interaction of positive and negative regulators. Centriolar coiled coil protein 110 (CP110) caps the distal end of the mother centriole and is known to act as a suppressor to control the timing of ciliogenesis. Here, we demonstrate that CP110 promotes cilia formation in vivo, in contrast to findings in cultured cells. Cp110−/− mice die shortly after birth owing to organogenesis defects as in ciliopathies. Shh signaling is impaired in null embryos and primary cilia are reduced in multiple tissues. We show that CP110 is required for anchoring of basal bodies to the membrane during cilia formation. CP110 loss resulted in an abnormal distribution of core components of subdistal appendages (SDAs) and of recycling endosomes, which may be associated with premature extension of axonemal microtubules. Our data implicate CP110 in SDA assembly and ciliary vesicle docking, two requisite early steps in cilia formation. We suggest that CP110 has unique context-dependent functions, acting as both a suppressor and a promoter of ciliogenesis. PMID:26965371

  16. An in vitro assay for entry into cilia reveals unique properties of the soluble diffusion barrier.

    PubMed

    Breslow, David K; Koslover, Elena F; Seydel, Federica; Spakowitz, Andrew J; Nachury, Maxence V

    2013-10-14

    Specific proteins are concentrated within primary cilia, whereas others remain excluded. To understand the mechanistic basis of entry into cilia, we developed an in vitro assay using cells in which the plasma membrane was permeabilized, but the ciliary membrane was left intact. Using a diffusion-to-capture system and quantitative analysis, we find that proteins >9 nm in diameter (∼100 kD) are restricted from entering cilia, and we confirm these findings in vivo. Interference with the nuclear pore complex (NPC) or the actin cytoskeleton in permeabilized cells demonstrated that the ciliary diffusion barrier is mechanistically distinct from those of the NPC or the axon initial segment. Moreover, applying a mass transport model to this system revealed diffusion coefficients for soluble and membrane proteins within cilia that are compatible with rapid exploration of the ciliary space in the absence of active transport. Our results indicate that large proteins require active transport for entry into cilia but not necessarily for movement inside cilia. PMID:24100294

  17. An in vitro assay for entry into cilia reveals unique properties of the soluble diffusion barrier

    PubMed Central

    Breslow, David K.; Koslover, Elena F.; Seydel, Federica; Spakowitz, Andrew J.

    2013-01-01

    Specific proteins are concentrated within primary cilia, whereas others remain excluded. To understand the mechanistic basis of entry into cilia, we developed an in vitro assay using cells in which the plasma membrane was permeabilized, but the ciliary membrane was left intact. Using a diffusion-to-capture system and quantitative analysis, we find that proteins >9 nm in diameter (∼100 kD) are restricted from entering cilia, and we confirm these findings in vivo. Interference with the nuclear pore complex (NPC) or the actin cytoskeleton in permeabilized cells demonstrated that the ciliary diffusion barrier is mechanistically distinct from those of the NPC or the axon initial segment. Moreover, applying a mass transport model to this system revealed diffusion coefficients for soluble and membrane proteins within cilia that are compatible with rapid exploration of the ciliary space in the absence of active transport. Our results indicate that large proteins require active transport for entry into cilia but not necessarily for movement inside cilia. PMID:24100294

  18. Manipulating Cilia Using the 3DFM

    NASA Astrophysics Data System (ADS)

    Fisher, Jay K.; O'Brien, E. Timothy; Taylor, R. M.; Davis, C. W.; Matsui, H.; Vicci, L.; Matthews, G.; Cribb, J.; Desai, K.; Wilde, B.; Superfine, R.

    2003-11-01

    Mucus flow generated by beating cilia projecting from epithelial cells is responsible for the removal of pathogens in the lungs. When the hydrodynamics of this mucociliary clearance system fails, as happens in the condition of Cystic Fibrosis, ensuing infections can destroy the lungs. A complex phenomenology includes the force generation of the cilia, the coupling of the cilia tips to the overlying mucus during the power stroke, and finally, it is speculated that the cilia act as force sensors to allow the control of the mucus viscosity and volume. We have designed a system that allows the measurement of 3 dimensional forces within an optical microscope using magnetic forces and superparamagnetic beads. We have functionalized the beads with antibodies and attached them to the cilia of human lung cell cultures. A laser is focused onto the bead whose motion is tracked using back focal plane detection of the forward scattered light. We have measured the 3 dimensional trajectory of beads attached to cilia beating up to 15Hz in human lung cell cultures with 10 nm, 1msec resolution. When forces are applied to the bead, we observe changes in the trajectory and velocity, implying a strong mechanoresponse of the cell as sensed by the cilia.

  19. Reduced cilia frequencies in human renal cell carcinomas versus neighboring parenchymal tissue

    PubMed Central

    2013-01-01

    eighty-nine clear cell, eight papillary, five chromophobe renal cell carcinomas, two sarcomatoid renal tumors and six oncocytomas were determined. A marked decrease of primary cilia across renal cell carcinoma subtypes was observed compared to adjacent nontumorigenic tissue. Conclusions Our study shows that cilia are predominantly lost in renal cell carcinomas compared to tissue of the tumor parenchyma. These results suggest that ciliary loss is common in renal tumorigenesis, possibly participating in the sequence of cellular events leading to malignant tumor development. Future therapies aimed at restoring or circumventing cilia signaling might therefore aid in current treatment efficacy. PMID:23369289

  20. Specialized Cilia in Mammalian Sensory Systems

    PubMed Central

    Falk, Nathalie; Lösl, Marlene; Schröder, Nadja; Gießl, Andreas

    2015-01-01

    Cilia and flagella are highly conserved and important microtubule-based organelles that project from the surface of eukaryotic cells and act as antennae to sense extracellular signals. Moreover, cilia have emerged as key players in numerous physiological, developmental, and sensory processes such as hearing, olfaction, and photoreception. Genetic defects in ciliary proteins responsible for cilia formation, maintenance, or function underlie a wide array of human diseases like deafness, anosmia, and retinal degeneration in sensory systems. Impairment of more than one sensory organ results in numerous syndromic ciliary disorders like the autosomal recessive genetic diseases Bardet-Biedl and Usher syndrome. Here we describe the structure and distinct functional roles of cilia in sensory organs like the inner ear, the olfactory epithelium, and the retina of the mouse. The spectrum of ciliary function in fundamental cellular processes highlights the importance of elucidating ciliopathy-related proteins in order to find novel potential therapies. PMID:26378583

  1. Cilia driven flow networks in the brain

    NASA Astrophysics Data System (ADS)

    Wang, Yong; Faubel, Regina; Westendorf, Chrsitian; Eichele, Gregor; Bodenschatz, Eberhard

    Neurons exchange soluble substances via the cerebrospinal fluid (CSF) that fills the ventricular system. The walls of the ventricular cavities are covered with motile cilia that constantly beat and thereby induce a directional flow. We recently discovered that cilia in the third ventricle generate a complex flow pattern leading to partitioning of the ventricular volume and site-directed transport paths along the walls. Transient and daily recurrent alterations in the cilia beating direction lead to changes in the flow pattern. This has consequences for delivery of CSF components along the near wall flow. The contribution of this cilia-induced flow to overall CSF flow remains to be investigated. The state-of-art lattice Boltzmann method is adapted for studying the CFS flow. The 3D geometry of the third ventricle at high resolution was reconstructed. Simulation of CSF flow without cilia in this geometry confirmed that the previous idea about unidirectional flow does not explain how different components of CSF can be delivered to their various target sites. We study the contribution of the cilia-induced flow pattern to overall CSF flow and identify target areas for site-specific delivery of CSF-constituents with respect to the temporal changes.

  2. DNAH11 Localization in the Proximal Region of Respiratory Cilia Defines Distinct Outer Dynein Arm Complexes.

    PubMed

    Dougherty, Gerard W; Loges, Niki T; Klinkenbusch, Judith A; Olbrich, Heike; Pennekamp, Petra; Menchen, Tabea; Raidt, Johanna; Wallmeier, Julia; Werner, Claudius; Westermann, Cordula; Ruckert, Christian; Mirra, Virginia; Hjeij, Rim; Memari, Yasin; Durbin, Richard; Kolb-Kokocinski, Anja; Praveen, Kavita; Kashef, Mohammad A; Kashef, Sara; Eghtedari, Fardin; Häffner, Karsten; Valmari, Pekka; Baktai, György; Aviram, Micha; Bentur, Lea; Amirav, Israel; Davis, Erica E; Katsanis, Nicholas; Brueckner, Martina; Shaposhnykov, Artem; Pigino, Gaia; Dworniczak, Bernd; Omran, Heymut

    2016-08-01

    Primary ciliary dyskinesia (PCD) is a recessively inherited disease that leads to chronic respiratory disorders owing to impaired mucociliary clearance. Conventional transmission electron microscopy (TEM) is a diagnostic standard to identify ultrastructural defects in respiratory cilia but is not useful in approximately 30% of PCD cases, which have normal ciliary ultrastructure. DNAH11 mutations are a common cause of PCD with normal ciliary ultrastructure and hyperkinetic ciliary beating, but its pathophysiology remains poorly understood. We therefore characterized DNAH11 in human respiratory cilia by immunofluorescence microscopy (IFM) in the context of PCD. We used whole-exome and targeted next-generation sequence analysis as well as Sanger sequencing to identify and confirm eight novel loss-of-function DNAH11 mutations. We designed and validated a monoclonal antibody specific to DNAH11 and performed high-resolution IFM of both control and PCD-affected human respiratory cells, as well as samples from green fluorescent protein (GFP)-left-right dynein mice, to determine the ciliary localization of DNAH11. IFM analysis demonstrated native DNAH11 localization in only the proximal region of wild-type human respiratory cilia and loss of DNAH11 in individuals with PCD with certain loss-of-function DNAH11 mutations. GFP-left-right dynein mice confirmed proximal DNAH11 localization in tracheal cilia. DNAH11 retained proximal localization in respiratory cilia of individuals with PCD with distinct ultrastructural defects, such as the absence of outer dynein arms (ODAs). TEM tomography detected a partial reduction of ODAs in DNAH11-deficient cilia. DNAH11 mutations result in a subtle ODA defect in only the proximal region of respiratory cilia, which is detectable by IFM and TEM tomography. PMID:26909801

  3. Developmental Signaling: Does It Bridge the Gap Between Cilia Dysfunction and Renal Cystogenesis?

    PubMed Central

    Tran, Pamela V.; Sharma, Madhulika; Li, Xiaogang; Calvet, James P.

    2015-01-01

    For more than a decade, evidence has accumulated linking dysfunction of primary cilia to renal cystogenesis, yet molecular mechanisms remain undefined. The pathogenesis of renal cysts is complex, involving multiple cellular aberrations and signaling pathways. Adding to this complexity, primary cilia exhibit multiple roles in a context-dependent manner. On renal epithelial cells, primary cilia act as mechanosensors and trigger extracellular Ca2+ influx in response to laminar fluid flow. During mammalian development, primary cilia mediate the Hedgehog (Hh), Wnt, and Notch pathways, which control cell proliferation and differentiation, and tissue morphogenesis. Further, experimental evidence suggests the developmental state of the kidney strongly influences renal cystic disease. Thus, we review evidence for regulation of Ca2+ and cAMP, key molecules in renal cystogenesis, at the primary cilium, the role of Hh, Wnt, and Notch signaling in renal cystic disease, and the interplay between these developmental pathways and Ca2+ signaling. Indeed if these developmental pathways influence renal cystogenesis, these may represent novel therapeutic targets that can be integrated into a combination therapy for renal cystic disease. PMID:24861210

  4. The zebrafish foxj1a transcription factor regulates cilia function in response to injury and epithelial stretch.

    PubMed

    Hellman, Nathan E; Liu, Yan; Merkel, Erin; Austin, Christina; Le Corre, Stephanie; Beier, David R; Sun, Zhaoxia; Sharma, Neeraj; Yoder, Bradley K; Drummond, Iain A

    2010-10-26

    Cilia are essential for normal organ function and developmental patterning, but their role in injury and regeneration responses is unknown. To probe the role of cilia in injury, we analyzed the function of foxj1, a transcriptional regulator of cilia genes, in response to tissue damage and renal cyst formation. Zebrafish foxj1a, but not foxj1b, was rapidly induced in response to epithelial distension and stretch, kidney cyst formation, acute kidney injury by gentamicin, and crush injury in spinal cord cells. Obstruction-induced up-regulation of foxj1a was not inhibited by cycloheximide, identifying foxj1a as a primary response gene to epithelial injury. Foxj1 was also dramatically up-regulated in murine cystic kidney disease epithelia [jck/jck (nek8) and Ift88Tg737Rpw(-/-)] as well as in response to kidney ischemia-reperfusion injury. Obstruction of the zebrafish pronephric tubule caused a rapid increase in cilia beat rate that correlated tightly with expanded tubule diameter and epithelial stretch. Zebrafish foxj1a was specifically required for cilia motility. Enhanced foxj1a expression in obstructed tubules induced cilia motility target genes efhc1, tektin-1, and dnahc9. foxj1a-deficient embryos failed to up-regulate efhc1, tektin-1, and dnahc9 and could not maintain enhanced cilia beat rates after obstruction, identifying an essential role for foxj1 in modulating cilia function after injury. These studies reveal that activation of a Foxj1 transcriptional network of ciliogenic genes is an evolutionarily conserved response to multiple forms of tissue damage and highlight enhanced cilia function as a previously uncharacterized component of organ homeostasis. PMID:20937855

  5. Sperm-Associated Antigen–17 Gene Is Essential for Motile Cilia Function and Neonatal Survival

    PubMed Central

    Teves, Maria Eugenia; Zhang, Zhibing; Costanzo, Richard M.; Henderson, Scott C.; Corwin, Frank D.; Zweit, Jamal; Sundaresan, Gobalakrishnan; Subler, Mark; Salloum, Fadi N.; Rubin, Bruce K.

    2013-01-01

    Primary ciliary dyskinesia (PCD), resulting from defects in cilia assembly or motility, is caused by mutations in a number of genes encoding axonemal proteins. PCD phenotypes are variable, and include recurrent respiratory tract infections, bronchiectasis, hydrocephaly, situs inversus, and male infertility. We generated knockout mice for the sperm-associated antigen–17 (Spag17) gene, which encodes a central pair (CP) protein present in the axonemes of cells with “9 + 2” motile cilia or flagella. The targeting of Spag17 resulted in a severe phenotype characterized by immotile nasal and tracheal cilia, reduced clearance of nasal mucus, profound respiratory distress associated with lung fluid accumulation and disruption of the alveolar epithelium, cerebral ventricular expansion consistent with emerging hydrocephalus, failure to suckle, and neonatal demise within 12 hours of birth. Ultrastructural analysis revealed the loss of one CP microtubule in approximately one quarter of tracheal cilia axonemes, an absence of a C1 microtubule projection, and other less frequent CP structural abnormalities. SPAG6 and SPAG16 (CP proteins that interact with SPAG17) were increased in tracheal tissue from SPAG17-deficient mice. We conclude that Spag17 plays a critical role in the function and structure of motile cilia, and that neonatal lethality is likely explained by impaired airway mucociliary clearance. PMID:23418344

  6. The essential roles of transition fibers in the context of cilia

    PubMed Central

    Wei, Qing; Ling, Kun; Hu, Jinghua

    2015-01-01

    Once thought of as a vestigial organelle, the primary cilium is now recognized as a signaling hub for key cellular pathways in vertebrate development. The recent renaissance in cilia studies significantly improved our understanding of how cilia form and function, but little is known about how ciliogenesis is initiated and how ciliary proteins enter cilia. These important ciliary events require transition fibers (TFs) that are positioned at the ciliary base as symmetric nine-bladed propeller fibrous structures. Up until recently, TFs have been the most underappreciated ciliary structures due to limited knowledge about their molecular composition and function. Here, we highlight recent advances in our understanding of TF composition and the indispensable roles of TFs in regulating the initiation of ciliogenesis and the selective import of ciliary proteins. PMID:25988548

  7. Mutations in the Cilia Gene ARL13B Lead to the Classical Form of Joubert Syndrome

    PubMed Central

    Cantagrel, Vincent; Silhavy, Jennifer L.; Bielas, Stephanie L.; Swistun, Dominika; Marsh, Sarah E.; Bertrand, Julien Y.; Audollent, Sophie; Attié-Bitach, Tania; Holden, Kenton R.; Dobyns, William B.; Traver, David; Al-Gazali, Lihadh; Ali, Bassam R.; Lindner, Tom H.; Caspary, Tamara; Otto, Edgar A.; Hildebrandt, Friedhelm; Glass, Ian A.; Logan, Clare V.; Johnson, Colin A.; Bennett, Christopher; Brancati, Francesco; Valente, Enza Maria; Woods, C. Geoffrey; Gleeson, Joseph G.

    2008-01-01

    Joubert syndrome (JS) and related disorders are a group of autosomal-recessive conditions sharing the “molar tooth sign” on axial brain MRI, together with cerebellar vermis hypoplasia, ataxia, and psychomotor delay. JS is suggested to be a disorder of cilia function and is part of a spectrum of disorders involving retinal, renal, digital, oral, hepatic, and cerebral organs. We identified mutations in ARL13B in two families with the classical form of JS. ARL13B belongs to the Ras GTPase family, and in other species is required for ciliogenesis, body axis formation, and renal function. The encoded Arl13b protein was expressed in developing murine cerebellum and localized to the cilia in primary neurons. Overexpression of human wild-type but not patient mutant ARL13B rescued the Arl13b scorpion zebrafish mutant. Thus, ARL13B has an evolutionarily conserved role mediating cilia function in multiple organs. PMID:18674751

  8. Stall Force and Response of Lung Cilia

    NASA Astrophysics Data System (ADS)

    Superfine, Richard; Hill, David; Swaminathan, Vinay; O'Brien, E. Timothy; Boucher, Ric; Button, Brian; Estes, Ashley

    2008-03-01

    We report on the response of lung cilia to applied forces. We have applied magnetic forces to magnetic beads attached to individual human lung cilia in cell cultures. Our magnetic system is capable of generating large forces (˜1nanoNewton on 1 micron beads) with a 3kHz bandwidth. We record the cilia beat motion using video microscopy to record beat frequency and amplitude as a function of applied force. We present three major findings. First, the stall force is approximately 150 pN. Second the frequency is unchanged by the application of forces up to the stall point. Third, the speed of the beat motion slows down according to the diminution of the beat amplitude while maintaining a constant frequency and the speed of the motion is the same whether the beat direction is in the same direction as the applied force or against the applied force.

  9. RFX3 governs growth and beating efficiency of motile cilia in mouse and controls the expression of genes involved in human ciliopathies.

    PubMed

    El Zein, Loubna; Ait-Lounis, Aouatef; Morlé, Laurette; Thomas, Joëlle; Chhin, Brigitte; Spassky, Nathalie; Reith, Walter; Durand, Bénédicte

    2009-09-01

    Cilia are cellular organelles that play essential physiological and developmental functions in various organisms. They can be classified into two categories, primary cilia and motile cilia, on the basis of their axonemal architecture. Regulatory factor X (RFX) transcription factors have been shown to be involved in the assembly of primary cilia in Caenorhabditis elegans, Drosophila and mice. Here, we have taken advantage of a novel primary-cell culture system derived from mouse brain to show that RFX3 is also necessary for biogenesis of motile cilia. We found that the growth and beating efficiencies of motile cilia are impaired in multiciliated Rfx3(-/-) cells. RFX3 was required for optimal expression of the FOXJ1 transcription factor, a key player in the differentiation program of motile cilia. Furthermore, we demonstrate for the first time that RFX3 regulates the expression of axonemal dyneins involved in ciliary motility by binding directly to the promoters of their genes. In conclusion, RFX proteins not only regulate genes involved in ciliary assembly, but also genes that are involved in ciliary motility and that are associated with ciliopathies such as primary ciliary dyskinesia in humans. PMID:19671664

  10. Mathematical embryology: the fluid mechanics of nodal cilia

    NASA Astrophysics Data System (ADS)

    Smith, D. J.; Smith, A. A.; Blake, J. R.

    2011-07-01

    Left-right symmetry breaking is critical to vertebrate embryonic development; in many species this process begins with cilia-driven flow in a structure termed the `node'. Primary `whirling' cilia, tilted towards the posterior, transport morphogen-containing vesicles towards the left, initiating left-right asymmetric development. We review recent theoretical models based on the point-force stokeslet and point-torque rotlet singularities, explaining how rotation and surface-tilt produce directional flow. Analysis of image singularity systems enforcing the no-slip condition shows how tilted rotation produces a far-field `stresslet' directional flow, and how time-dependent point-force and time-independent point-torque models are in this respect equivalent. Associated slender body theory analysis is reviewed; this approach enables efficient and accurate simulation of three-dimensional time-dependent flow, time-dependence being essential in predicting features of the flow such as chaotic advection, which have subsequently been determined experimentally. A new model for the nodal flow utilising the regularized stokeslet method is developed, to model the effect of the overlying Reichert's membrane. Velocity fields and particle paths within the enclosed domain are computed and compared with the flow profiles predicted by previous `membrane-less' models. Computations confirm that the presence of the membrane produces flow-reversal in the upper region, but no continuous region of reverse flow close to the epithelium. The stresslet far-field is no longer evident in the membrane model, due to the depth of the cavity being of similar magnitude to the cilium length. Simulations predict that vesicles released within one cilium length of the epithelium are generally transported to the left via a `loopy drift' motion, sometimes involving highly unpredictable detours around leftward cilia [truncated

  11. Cottingham formula and nucleon polarisabilities

    NASA Astrophysics Data System (ADS)

    Gasser, J.; Hoferichter, M.; Leutwyler, H.; Rusetsky, A.

    2015-08-01

    The difference between the electromagnetic self-energies of proton and neutron can be calculated with the Cottingham formula, which expresses the self-energies as an integral over the electroproduction cross sections - provided the nucleon matrix elements of the current commutator do not contain a fixed pole. We show that, under the same proviso, the subtraction function occurring in the dispersive representation of the virtual Compton forward scattering amplitude is determined by the cross sections. The representation in particular leads to a parameter-free sum rule for the nucleon polarisabilities. We evaluate the sum rule for the difference between the electric polarisabilities of proton and neutron by means of the available parameterisations of the data and compare the result with experiment.

  12. Regeneration of cilia in heavily irradiated sea urchin embryos

    SciTech Connect

    Rustad, R.C.

    1981-12-01

    Cilia were removed from blastulae, gastrulae, and plutei of the sea urchins Arbacia punctulata and Lytechinus variegatus by shaking the embryos in hypertonic media. Exposure to 50 krad (and in some experiments 100 krad) of ..gamma.. radiation either before or after deciliation had no effect on the time of appearance of regenerating cilia. There were no visually obvious differences in the rate of growth of the cilia in control and irradiated embryos. The cilia commenced beating at the same time, but the initial beating sometimes seemed less vigorous following irradiation. The data support the hypothesis that radiation has no major effect on the assembly from mature basal bodies of the microtubules of cilia.

  13. Cildb: a knowledgebase for centrosomes and cilia

    PubMed Central

    Arnaiz, Olivier; Malinowska, Agata; Klotz, Catherine; Sperling, Linda; Dadlez, Michal; Koll, France; Cohen, Jean

    2009-01-01

    Ciliopathies, pleiotropic diseases provoked by defects in the structure or function of cilia or flagella, reflect the multiple roles of cilia during development, in stem cells, in somatic organs and germ cells. High throughput studies have revealed several hundred proteins that are involved in the composition, function or biogenesis of cilia. The corresponding genes are potential candidates for orphan ciliopathies. To study ciliary genes, model organisms are used in which particular questions on motility, sensory or developmental functions can be approached by genetics. In the course of high throughput studies of cilia in Paramecium tetraurelia, we were confronted with the problem of comparing our results with those obtained in other model organisms. We therefore developed a novel knowledgebase, Cildb, that integrates ciliary data from heterogeneous sources. Cildb links orthology relationships among 18 species to high throughput ciliary studies, and to OMIM data on human hereditary diseases. The web interface of Cildb comprises three tools, BioMart for complex queries, BLAST for sequence homology searches and GBrowse for browsing the human genome in relation to OMIM information for human diseases. Cildb can be used for interspecies comparisons, building candidate ciliary proteomes in any species, or identifying candidate ciliopathy genes. Database URL: http://cildb.cgm.cnrs-gif.fr PMID:20428338

  14. Transmembrane protein OSTA-1 shapes sensory cilia morphology via regulation of intracellular membrane trafficking in C. elegans

    PubMed Central

    Olivier-Mason, Anique; Wojtyniak, Martin; Bowie, Rachel V.; Nechipurenko, Inna V.; Blacque, Oliver E.; Sengupta, Piali

    2013-01-01

    The structure and function of primary cilia are critically dependent on intracellular trafficking pathways that transport ciliary membrane and protein components. The mechanisms by which these trafficking pathways are regulated are not fully characterized. Here we identify the transmembrane protein OSTA-1 as a new regulator of the trafficking pathways that shape the morphology and protein composition of sensory cilia in C. elegans. osta-1 encodes an organic solute transporter alpha-like protein, mammalian homologs of which have been implicated in membrane trafficking and solute transport, although a role in regulating cilia structure has not previously been demonstrated. We show that mutations in osta-1 result in altered ciliary membrane volume, branch length and complexity, as well as defects in localization of a subset of ciliary transmembrane proteins in different sensory cilia types. OSTA-1 is associated with transport vesicles, localizes to a ciliary compartment shown to house trafficking proteins, and regulates both retrograde and anterograde flux of the endosome-associated RAB-5 small GTPase. Genetic epistasis experiments with sensory signaling, exocytic and endocytic proteins further implicate OSTA-1 as a crucial regulator of ciliary architecture via regulation of cilia-destined trafficking. Our findings suggest that regulation of transport pathways in a cell type-specific manner contributes to diversity in sensory cilia structure and might allow dynamic remodeling of ciliary architecture via multiple inputs. PMID:23482491

  15. Entropy-based measures of in vivo cilia-driven microfluidic mixing derived from quantitative optical imaging

    NASA Astrophysics Data System (ADS)

    Chandrasekera, Kenny; Jonas, Stephan; Bhattacharya, Dipankan; Khokha, Mustafa; Choma, Michael A.

    2012-02-01

    Motile cilia are cellular organelles that project from different epithelial surfaces including respiratory epithelium. They generate directional fluid flow that removes harmful pathogens and particulate matter from the respiratory system. While it has been known that primary ciliary dyskinesia increases the risk of recurrent pulmonary infections, there is now heightened interest in understanding the role that cilia play in a wide-variety of respiratory diseases. Different optical imaging technologies are being investigated to visualize cilia-driven fluid flow, and quantitative image analysis is used to generate measures of ciliary performance. Here, we demonstrate the quantification of in vivo cilia-driven microfluidic mixing using spatial and temporal measures of Shannon information entropy. Using videomicroscopy, we imaged in vivo cilia-driven fluid flow generated by the epidermis of the Xenopus tropicalis embryo. Flow was seeded with either dyes or microparticles. Both spatial and temporal measures of entropy show significant levels of mixing, with maximum entropy measures of ~6.5 (out of a possible range of 0 to 8). Spatial entropy measures showed localization of mixing "hot-spots" and "cold-spots" and temporal measures showed mixing throughout.In sum, entropy-based measures of microfluidic mixing can characterize in vivo cilia-driven fluid flow and hold the potential for better characterization of ciliary dysfunction.

  16. Bio-inspired artificial cilia with magnetic dynamic properties

    NASA Astrophysics Data System (ADS)

    Sun, Leilei; Zheng, Yongmei

    2015-04-01

    Inspired by the structure and properties of natural cilia, we focused on a facile template-free approach to prepare magnetic artificial cilia grown on the substrate (glass, PDMS, or others). In an applied magnetic field, the cilia formed spontaneously and immediately from magnetic nanoparticles and elastomeric polymer in a liquid solvent by bottom-up self-assembly. The length of prepared cilia could be in the scale of millimeter and reach a high aspect ratio of even over 100. We studied the effect of the magnetic strength applied and the size of nanoparticles to get tunable scale of cilia. The cilia show reversibly bending in an external magnetic field and this bending actuation gave some important functions: to transport macroscopic nonmagnetic materials on the cilia and to mix liquids.

  17. Microtubule modifications and stability are altered by cilia perturbation and in cystic kidney disease

    PubMed Central

    Berbari, Nicolas F.; Sharma, Neeraj; Malarkey, Erik B.; Pieczynski, Jay N.; Boddu, Ravindra; Gaertig, Jacek; Guay-Woodford, Lisa; Yoder, Bradley K.

    2013-01-01

    Summary Disruption of the primary cilium is associated with a growing number of human diseases collectively termed ciliopathies. Ciliopathies present with a broad range of clinical features consistent with the near ubiquitous nature of the organelle and its role in diverse signaling pathways throughout development and adult homeostasis. The clinical features associated with cilia dysfunction can include such phenotypes as polycystic kidneys, skeletal abnormalities, blindness, anosmia, and obesity. Although the clinical relevance of the primary cilium is evident, the effects that cilia dysfunction has on the cell and how this contributes to disease remains poorly understood. Here, we show that loss of ciliogenesis genes such as Ift88 and Kif3a lead to increases in post-translational modifications on cytosolic microtubules. This effect was observed in cilia mutant kidney cells grown in vitro and in vivo in cystic kidneys. The hyper-acetylation of microtubules resulting from cilia loss is associated with both altered microtubule stability and increased α-tubulin acetyl-transferase activity. Intriguingly, the effect on microtubules was also evident in renal samples from patients with autosomal recessive polycystic kidneys. These findings indicate that altered microtubule post-translational modifications may influence some of the phenotypes observed in ciliopathies. PMID:23124988

  18. IFT46 plays an essential role in cilia development

    PubMed Central

    Lee, Mi-Sun; Hwang, Kyu-Seok; Oh, Hyun-Woo; Ji-Ae, Kim; Kim, Hyun-Taek; Cho, Hyun-Soo; Lee, Jeong-Ju; Ko, Je Yeong; Choi, Jung-Hwa; Jeong, Yun-Mi; You, Kwan-Hee; Kim, Joon; Park, Doo-Sang; Nam, Ki-Hoan; Aizawa, Shinichi; Kiyonari, Hiroshi; Shioi, Go; Park, Jong-Hoon; Zhou, Weibin; Kim, Nam-Soon; Kim, Cheol-Hee

    2015-01-01

    Cilia are microtubule-based structures that project into the extracellular space. Ciliary defects are associated with several human diseases, including polycystic kidney disease, primary ciliary dyskinesia, left-right axis patterning, hydrocephalus and retinal degeneration. However, the genetic and cellular biological control of ciliogenesis remains poorly understood. The IFT46 is one of the highly conserved intraflagellar transport complex B proteins. In zebrafish, ift46 is expressed in various ciliated tissues such as Kupffer’s vesicle, pronephric ducts, ears and spinal cord. We show that ift46 is localized to the basal body. Knockdown of ift46 gene results in multiple phenotypes associated with various ciliopathies including kidney cysts, pericardial edema and ventral axis curvature. In ift46 morphants, cilia in kidney and spinal canal are shortened and abnormal. Similar ciliary defects are observed in otic vesicles, lateral line hair cells, olfactory pits, but not in Kupffer’s vesicle. To explore the functions of Ift46 during mouse development, we have generated Ift46 knock-out mice. The Ift46 mutants have developmental defects in brain, neural tube and heart. In particular Ift46(−/−) homozygotes displays randomization of the embryo heart looping, which is a hallmark of defective left-right (L/R) axis patterning. Taken together, our results demonstrated that IFT46 has an essential role in vertebrate ciliary development. PMID:25722189

  19. Magnetically-actuated artificial cilia for microfluidic propulsion.

    PubMed

    Khaderi, S N; Craus, C B; Hussong, J; Schorr, N; Belardi, J; Westerweel, J; Prucker, O; Rühe, J; den Toonder, J M J; Onck, P R

    2011-06-21

    In this paper we quantitatively analyse the performance of magnetically-driven artificial cilia for lab-on-a-chip applications. The artificial cilia are fabricated using thin polymer films with embedded magnetic nano-particles and their deformation is studied under different external magnetic fields and flows. A coupled magneto-mechanical solid-fluid model that accurately captures the interaction between the magnetic field, cilia and fluid is used to simulate the cilia motion. The elastic and magnetic properties of the cilia are obtained by fitting the results of the computational model to the experimental data. The performance of the artificial cilia with a non-uniform cross-section is characterised using the numerical model for two channel configurations that are of practical importance: an open-loop and a closed-loop channel. We predict that the flow and pressure head generated by the artificial cilia can be as high as 18 microlitres per minute and 3 mm of water, respectively. We also study the effect of metachronal waves on the flow generated and show that the fluid propelled increases drastically compared to synchronously beating cilia, and is unidirectional. This increase is significant even when the phase difference between adjacent cilia is small. The obtained results provide guidelines for the optimal design of magnetically-driven artificial cilia for microfluidic propulsion. PMID:21331419

  20. Polarised antibranes from Smarr relations

    NASA Astrophysics Data System (ADS)

    Cohen-Maldonado, Diego; Diaz, Juan; Gautason, Fridrik Freyr

    2016-05-01

    We study the backreaction of smeared and localised anti M2-branes placed at the tip of the CGLP background. To this end we derive a Smarr relation for backreacted antibranes at zero and finite temperature. For extremal antibranes we show that if smeared they cannot have regular horizons, whereas localised M2-branes can potentially be regular when polarised into M5-branes, in agreement with the probe result of Klebanov and Pufu. We further discuss antibranes at finite temperature and argue that localised antibrane solutions with regular horizons are not excluded.

  1. Transport of the outer dynein arm complex to cilia requires a cytoplasmic protein Lrrc6.

    PubMed

    Inaba, Yasuko; Shinohara, Kyosuke; Botilde, Yanick; Nabeshima, Ryo; Takaoka, Katsuyoshi; Ajima, Rieko; Lamri, Lynda; Takeda, Hiroyuki; Saga, Yumiko; Nakamura, Tetsuya; Hamada, Hiroshi

    2016-07-01

    Lrrc6 encodes a cytoplasmic protein that is expressed specifically in cells with motile cilia including the node, trachea and testes of the mice. A mutation of Lrrc6 has been identified in human patients with primary ciliary dyskinesia (PCD). Mutant mice lacking Lrrc6 show typical PCD defects such as hydrocephalus and laterality defects. We found that in the absence of Lrrc6, the morphology of motile cilia remained normal, but their motility was completely lost. The 9 + 2 arrangement of microtubules remained normal in Lrrc6(-/-) mice, but the outer dynein arms (ODAs), the structures essential for the ciliary beating, were absent from the cilia. In the absence of Lrrc6, ODA proteins such as DNAH5, DNAH9 and IC2, which are assembled in the cytoplasm and transported to the ciliary axoneme, remained in the cytoplasm and were not transported to the ciliary axoneme. The IC2-IC1 interaction, which is the first step of ODA assembly, was normal in Lrrc6(-/-) mice testes. Our results suggest that ODA proteins may be transported from the cytoplasm to the cilia by an Lrrc6-dependent mechanism. PMID:27353389

  2. A simple homemade polarised sunglasses test card

    NASA Astrophysics Data System (ADS)

    Bamdad, Farzad

    2016-05-01

    In this article construction of a simple and inexpensive test card which can be used to demonstrate the polarisation ability of sunglasses is described. The card was fabricated simply by using a piece of polariser sheet with one to three layers of cellophane tape fixed on it.

  3. A Simple Homemade Polarised Sunglasses Test Card

    ERIC Educational Resources Information Center

    Bamdad, Farzad

    2016-01-01

    In this article construction of a simple and inexpensive test card which can be used to demonstrate the polarisation ability of sunglasses is described. The card was fabricated simply by using a piece of polariser sheet with one to three layers of cellophane tape fixed on it.

  4. Polarisation of Social Studies Textbooks in Pakistan

    ERIC Educational Resources Information Center

    Zaidi, Syed Manzar Abbas

    2011-01-01

    This article looks at the evolution of the social studies curricula in Pakistan, which are of critical importance in shaping the outlook of many young Pakistanis, who are affected by this polarised discourse. The author argues that this trend of polarisation springing from dynamics of education also effectively contributes to a widening social…

  5. Assembly and dynamics of synthetic cilia

    NASA Astrophysics Data System (ADS)

    Sanchez, Tim

    2012-02-01

    From motility of simple protists to determining the handedness of complex vertebrates, highly conserved eukaryotic cilia and flagella are essential for the reproduction and survival of many biological organisms. Despite extensive studies, the exact mechanism by which individual components coordinate to produce ciliary beating patterns remains unknown. We describe a novel approach towards studying ciliary beating. Instead of deconstructing a fully functional organelle from the top-down, we describe a process by which synthetic cilia-like structures are assembled from the bottom-up. We find that simple mixtures of microtubules, kinesin clusters, and a bundling agent produce spontaneous oscillations in MT bundles, suggesting that self-organized beating may be a generic feature of internally driven bundles. Furthermore, bundles in close proximity spontaneously coordinate their beating to generate metachronal traveling waves, reminiscent of the waves seen in ciliary fields. These findings and future refinements of the system can potentially provide insights into general design principles required for engineering synthetic cilia as well as understanding the biological analogues.

  6. Bardet-Biedl syndrome: Is it only cilia dysfunction?

    PubMed

    Novas, Rossina; Cardenas-Rodriguez, Magdalena; Irigoín, Florencia; Badano, Jose L

    2015-11-14

    Bardet-Biedl syndrome (BBS) is a genetically heterogeneous, pleiotropic disorder, characterized by both congenital and late onset defects. From the analysis of the mutational burden in patients to the functional characterization of the BBS proteins, this syndrome has become a model for both understanding oligogenic patterns of inheritance and the biology of a particular cellular organelle: the primary cilium. Here we briefly review the genetics of BBS to then focus on the function of the BBS proteins, not only in the context of the cilium but also highlighting potential extra-ciliary roles that could be relevant to the etiology of the disorder. Finally, we provide an overview of how the study of this rare syndrome has contributed to the understanding of cilia biology and how this knowledge has informed on the cellular basis of different clinical manifestations that characterize BBS and the ciliopathies. PMID:26231314

  7. Microfabrication of IPMC cilia for bio-inspired flow sensing

    NASA Astrophysics Data System (ADS)

    Lei, Hong; Li, Wen; Tan, Xiaobo

    2012-04-01

    As the primary flow sensing organ for fishes, the lateral line system plays a critical role in fish behavior. Analogous to its biological counterpart, an artificial lateral line system, consisting of arrays of micro flow sensors, is expected to be instrumental in the navigation and control of underwater robots. In this paper we investigate the microfabrication of ionic polymer-metal composite (IPMC) cilia for the purpose of flow sensing. While existing macro- and microfabrication methods for IPMCs have predominantly focused on planar structures, we propose a device where micro IPMC beams stand upright on a substrate to effectively interact with the flow. Challenges in the casting of 3D Nafion structure and selective formation of electrodes are discussed, and potential solutions for addressing these challenges are presented together with preliminary microfabrication results.

  8. Methods for imaging individual cilia in living echinoid embryos.

    PubMed

    Morris, Robert L; Pope, Hans W; Sholi, Adam N; Williams, Leah M; Ettinger, Chelsea R; Beacham, Gwendolyn M; Shintaku, Tatsushi; Abbott, Zachary D; Doherty, Elyse M

    2015-01-01

    The embryos of echinoids (sea urchins and sand dollars) serve as excellent models for studying cilia differentiation and stages of the cilia life cycle including ciliogenic initiation, growth, maintenance, and retraction. Early in echinoid development, uniform motile cilia form on all cells simultaneously but then rapidly differentiate into multiple cilia types that differ in morphology, motility, and signaling sensitivity. Metal ion treatments that shift germ layer boundaries and thereby "animalize" or "vegetalize" embryos can be used to enrich for low-abundance cilia types rendering those specialized cilia and the differentiation processes they exhibit much easier to study. The experimental advantages of having robust cilia growth and differentiation is tempered by the challenge of restraining ciliated embryos well enough to view the process of ciliogenesis live. We have developed four observation chambers as modifications of the Kiehart chamber for long-term light microscopic imaging of ciliated echinoid embryos. One of these systems employs paramagnetic beads to render ciliated larvae magnetic so they can be gently and reversibly trapped directly under the objective lens. With this magnetic trapping system, the larva can be positioned and repositioned until they achieve the orientation with the clearest view of any cilia of interest. These methods of gentle embryo restraint allow normal embryo development and the normal ciliogenic cycle and ciliary differentiation processes to continue in direct view. Sequential image series can then be collected and analyzed to quantitatively study the wide spectrum of cilia behaviors and properties that arise in developing echinoid embryos. PMID:25837394

  9. Ion channels and calcium signaling in motile cilia

    PubMed Central

    Doerner, Julia F; Delling, Markus; Clapham, David E

    2015-01-01

    The beating of motile cilia generates fluid flow over epithelia in brain ventricles, airways, and Fallopian tubes. Here, we patch clamp single motile cilia of mammalian ependymal cells and examine their potential function as a calcium signaling compartment. Resting motile cilia calcium concentration ([Ca2+] ~170 nM) is only slightly elevated over cytoplasmic [Ca2+] (~100 nM) at steady state. Ca2+ changes that arise in the cytoplasm rapidly equilibrate in motile cilia. We measured CaV1 voltage-gated calcium channels in ependymal cells, but these channels are not specifically enriched in motile cilia. Membrane depolarization increases ciliary [Ca2+], but only marginally alters cilia beating and cilia-driven fluid velocity within short (~1 min) time frames. We conclude that beating of ependymal motile cilia is not tightly regulated by voltage-gated calcium channels, unlike that of well-studied motile cilia and flagella in protists, such as Paramecia and Chlamydomonas. DOI: http://dx.doi.org/10.7554/eLife.11066.001 PMID:26650848

  10. Sensory functions of motile cilia and implication for bronchiectasis

    PubMed Central

    Jain, Raksha; Javidan-Nejad, Cylen; Alexander-Brett, Jennifer; Horani, Amjad; Cabellon, Michelle C.; Walter, Michael J.; Brody, Steven L.

    2013-01-01

    Cilia are specialized organelles that extend from the surface of cells into the local environment. Airway epithelial cell cilia are motile to provide mucociliary clearance for host defense. On other cells, solitary cilia are specialized to detect chemical or mechanosensory signals. Sensory proteins in motile cilia have recently been identified that detect shear stress, osmotic force, fluid flow, bitter taste and sex hormones. The relationship of sensory function in human motile cilia to disease is now being revealed. One example is polycystin-1 and polycystin-2. As a complex, these proteins function as a flow sensor in cilia and are mutated in autosomal dominant polycystic kidney disease (ADPKD). The polycystins are also expressed in motile cilia of the airways, potentially operating as sensors in the lung. Computed tomography studies from patients with ADPKD revealed radiographic evidence for bronchiectasis, suggesting that polycystin-1 and -2 are important in lung function. The expression of this complex and sensory channel TRPV4, and bitter taste and sex hormones receptors in motile cilia indicate that the cell is wired to interpret environmental cues to regulate cilia beat frequency and other functions. Defective signaling of sensory proteins may result in a ciliopathy that includes lung disease. PMID:22202111

  11. Methods for Studying Movement of Molecules Within Cilia.

    PubMed

    Lechtreck, Karl F

    2016-01-01

    The assembly of cilia and eukaryotic flagella (interchangeable terms) requires the import of numerous proteins from the cell body into the growing organelle. Proteins move into and inside cilia by diffusion and by motor-based intraflagellar transport (IFT). Many aspects of ciliary protein transport such as the distribution of unloading sites and the frequency of transport can be analyzed using direct in vivo imaging of fluorescently tagged proteins. Here, we will describe how to use total internal reflection fluorescence microcopy (TIRFM) to analyze protein transport in the flagella of the unicellular alga Chlamydomonas reinhardtii, a widely used model for cilia and cilia-related disease. PMID:27514917

  12. Vector solitons with a uniform polarisation state induced by polarisation filtering in a fibre laser

    SciTech Connect

    Chen Weicheng; Chen Guojie; Han Dingan; Xie Jianing

    2013-06-30

    A fibre laser with a SESAM as a passive mode-locker is constructed for obtaining a vector soliton with the Kelly sidebands. The analysis of the peculiarities of the sidebands shows that the polarisation states are nonuniform across the entire pulse spectral profile from the leading edge to the trailing edge. Polarisation filtering effect is proposed to obtain a vector soliton with a uniform polarisation state. It is shown that during the polarisation filtering by a polariser incorporated into the laser cavity, the spectral width of the vector solitons gradually broadens and the pulse power decreases. It is found that at a maximum spectral width and a minimum pulse power, vector solitons with a uniform polarisation state are generated. (nonlinear optical phenomena)

  13. Left-right asymmetry: cilia and calcium revisited.

    PubMed

    Blum, Martin; Vick, Philipp

    2015-03-01

    Leftward flow generated by motile cilia is known to underlie left-right asymmetry in vertebrate embryos. A new study now links intraciliary calcium oscillations to cilia motility and the downstream nodal signaling cascade that drives left-sided development. PMID:25734272

  14. Cilia and Nuclear Pore Proteins: Pore No More?

    PubMed

    Obado, Samson O; Rout, Michael P

    2016-09-12

    Nuclear pore proteins at the base of cilia were thought to regulate transport into cilia. In this issue of Developmental Cell, Del Viso et al. (2016) challenge this view, showing instead that pore proteins localize to ciliary basal bodies and that their perturbation leads to congenital heart disease. PMID:27623377

  15. ATR promotes cilia signalling: links to developmental impacts.

    PubMed

    Stiff, Tom; Casar Tena, Teresa; O'Driscoll, Mark; Jeggo, Penny A; Philipp, Melanie

    2016-04-15

    Mutations in ATR(ataxia telangiectasia and RAD3-related) cause Seckel syndrome (ATR-SS), a microcephalic primordial dwarfism disorder. Hitherto, the clinical manifestation of ATR deficiency has been attributed to its canonical role in DNA damage response signalling following replication fork stalling/collapse. Here, we show that ATR regulates cilia-dependent signalling in a manner that can be uncoupled from its function during replication. ATR-depleted or patient-derived ATR-SS cells form cilia of slightly reduced length but are dramatically impaired in cilia-dependent signalling functions, including growth factor and Sonic hedgehog signalling. To better understand the developmental impact of ATR loss of function, we also used zebrafish as a model. Zebrafish embryos depleted of Atr resembled ATR-SS morphology, showed a modest but statistically significant reduction in cilia length and other morphological features indicative of cilia dysfunction. Additionally, they displayed defects in left-right asymmetry including ambiguous expression of southpaw, incorrectly looped hearts and randomized localization of internal organs including the pancreas, features typically conferred by cilia dysfunction. Our findings reveal a novel role for ATR in cilia signalling distinct from its canonical function during replication and strengthen emerging links between cilia function and development. PMID:26908596

  16. ATR promotes cilia signalling: links to developmental impacts

    PubMed Central

    Stiff, Tom; Casar Tena, Teresa; O'Driscoll, Mark; Jeggo, Penny A.; Philipp, Melanie

    2016-01-01

    Mutations in ATR (ataxia telangiectasia and RAD3-related) cause Seckel syndrome (ATR-SS), a microcephalic primordial dwarfism disorder. Hitherto, the clinical manifestation of ATR deficiency has been attributed to its canonical role in DNA damage response signalling following replication fork stalling/collapse. Here, we show that ATR regulates cilia-dependent signalling in a manner that can be uncoupled from its function during replication. ATR-depleted or patient-derived ATR-SS cells form cilia of slightly reduced length but are dramatically impaired in cilia-dependent signalling functions, including growth factor and Sonic hedgehog signalling. To better understand the developmental impact of ATR loss of function, we also used zebrafish as a model. Zebrafish embryos depleted of Atr resembled ATR-SS morphology, showed a modest but statistically significant reduction in cilia length and other morphological features indicative of cilia dysfunction. Additionally, they displayed defects in left-right asymmetry including ambiguous expression of southpaw, incorrectly looped hearts and randomized localization of internal organs including the pancreas, features typically conferred by cilia dysfunction. Our findings reveal a novel role for ATR in cilia signalling distinct from its canonical function during replication and strengthen emerging links between cilia function and development. PMID:26908596

  17. First Polarised Light with the NIKA Camera

    NASA Astrophysics Data System (ADS)

    Ritacco, A.; Adam, R.; Adane, A.; Ade, P.; André, P.; Beelen, A.; Belier, B.; Benoît, A.; Bideaud, A.; Billot, N.; Bourrion, O.; Calvo, M.; Catalano, A.; Coiffard, G.; Comis, B.; D'Addabbo, A.; Désert, F.-X.; Doyle, S.; Goupy, J.; Kramer, C.; Leclercq, S.; Macías-Pérez, J. F.; Martino, J.; Mauskopf, P.; Maury, A.; Mayet, F.; Monfardini, A.; Pajot, F.; Pascale, E.; Perotto, L.; Pisano, G.; Ponthieu, N.; Rebolo-Iglesias, M.; Revéret, V.; Rodriguez, L.; Savini, G.; Schuster, K.; Sievers, A.; Thum, C.; Triqueneaux, S.; Tucker, C.; Zylka, R.

    2015-10-01

    NIKA is a dual-band camera operating with 315 frequency multiplexed LEKIDs cooled at 100 mK. NIKA is designed to observe the sky in intensity and polarisation at 150 and 260 GHz from the IRAM 30-m telescope. It is a test-bench for the final NIKA2 camera. The incoming linear polarisation is modulated at four times the mechanical rotation frequency by a warm rotating multi-layer half- wave plate. Then, the signal is analyzed by a wire grid and finally absorbed by the lumped element kinetic inductance detectors (LEKIDs). The small time constant (< 1 ms ) of the LEKIDs combined with the modulation of the HWP enables the quasi-simultaneous measurement of the three Stokes parameters I, Q, U, representing linear polarisation. In this paper, we present the results of recent observational campaigns demonstrating the good performance of NIKA in detecting polarisation at millimeter wavelength.

  18. First Polarised Light with the NIKA Camera

    NASA Astrophysics Data System (ADS)

    Ritacco, A.; Adam, R.; Adane, A.; Ade, P.; André, P.; Beelen, A.; Belier, B.; Benoît, A.; Bideaud, A.; Billot, N.; Bourrion, O.; Calvo, M.; Catalano, A.; Coiffard, G.; Comis, B.; D'Addabbo, A.; Désert, F.-X.; Doyle, S.; Goupy, J.; Kramer, C.; Leclercq, S.; Macías-Pérez, J. F.; Martino, J.; Mauskopf, P.; Maury, A.; Mayet, F.; Monfardini, A.; Pajot, F.; Pascale, E.; Perotto, L.; Pisano, G.; Ponthieu, N.; Rebolo-Iglesias, M.; Revéret, V.; Rodriguez, L.; Savini, G.; Schuster, K.; Sievers, A.; Thum, C.; Triqueneaux, S.; Tucker, C.; Zylka, R.

    2016-08-01

    NIKA is a dual-band camera operating with 315 frequency multiplexed LEKIDs cooled at 100 mK. NIKA is designed to observe the sky in intensity and polarisation at 150 and 260 GHz from the IRAM 30-m telescope. It is a test-bench for the final NIKA2 camera. The incoming linear polarisation is modulated at four times the mechanical rotation frequency by a warm rotating multi-layer half- wave plate. Then, the signal is analyzed by a wire grid and finally absorbed by the lumped element kinetic inductance detectors (LEKIDs). The small time constant (<1 ms ) of the LEKIDs combined with the modulation of the HWP enables the quasi-simultaneous measurement of the three Stokes parameters I, Q, U, representing linear polarisation. In this paper, we present the results of recent observational campaigns demonstrating the good performance of NIKA in detecting polarisation at millimeter wavelength.

  19. Polarisation losses in a ring prism cavity

    SciTech Connect

    Kuryatov, V N; Sokolov, A L

    2000-02-28

    The polarisation losses in a ring cavity, formed by total-internal-reflection prisms, were analysed. All the sources of the polarisation losses are indicated and expressions for their calculation are presented. The limit to loss reduction in cavities of this kind, set by the difference between the radii of curvature of the radiation wavefront and of the refracting prism faces, was determined. (laser gyroscopes)

  20. An ultrafast carbon nanotube terahertz polarisation modulator

    SciTech Connect

    Docherty, Callum J.; Stranks, Samuel D.; Habisreutinger, Severin N.; Joyce, Hannah J.; Herz, Laura M.; Nicholas, Robin J.; Johnston, Michael B.

    2014-05-28

    We demonstrate ultrafast modulation of terahertz radiation by unaligned optically pumped single-walled carbon nanotubes. Photoexcitation by an ultrafast optical pump pulse induces transient terahertz absorption in nanowires aligned parallel to the optical pump. By controlling the polarisation of the optical pump, we show that terahertz polarisation and modulation can be tuned, allowing sub-picosecond modulation of terahertz radiation. Such speeds suggest potential for semiconductor nanowire devices in terahertz communication technologies.

  1. An ultrafast carbon nanotube terahertz polarisation modulator

    NASA Astrophysics Data System (ADS)

    Docherty, Callum J.; Stranks, Samuel D.; Habisreutinger, Severin N.; Joyce, Hannah J.; Herz, Laura M.; Nicholas, Robin J.; Johnston, Michael B.

    2014-05-01

    We demonstrate ultrafast modulation of terahertz radiation by unaligned optically pumped single-walled carbon nanotubes. Photoexcitation by an ultrafast optical pump pulse induces transient terahertz absorption in nanowires aligned parallel to the optical pump. By controlling the polarisation of the optical pump, we show that terahertz polarisation and modulation can be tuned, allowing sub-picosecond modulation of terahertz radiation. Such speeds suggest potential for semiconductor nanowire devices in terahertz communication technologies.

  2. Dynamic nuclear polarisation of diamond

    NASA Astrophysics Data System (ADS)

    High, Grant Lysle

    Chapter one reviews the reported literature on the NMR of diamond. This signal consists of a single line at 39 ppm from TMS and two hyperfine lines due to 13C interactions. The second chapter introduces the apparatus used in this study. The availability of this excellently equipped laboratory presented a unique opportunity to perform this investigation. Chapter three outlines the experimental techniques used as well as the manner in which the acquired data was processed. The fourth chapter presents an overview of the most common defects found in diamond. Proposed models of these defects are presented and the EPR spectra displayed. The methods developed to determine the paramagnetic impurity concentration, from the EPR line width and the spin-spin relaxation times are presented in the fifth chapter. The line width gives the total paramagnetic impurity concentration to about 10 ppm. The spin-spin relaxation time allows the determination of paramagnetic impurity concentrations, to much lower levels. This information was used in the explanation of the relaxation behaviour for the diamonds investigated. The temperature dependence of the paramagnetic electron relaxation times is reported in the sixth chapter. The results obtained are consistent with the findings in prior work that P1 impurities are typical, Jahn Teller centres. Two diamonds, however, display trends that depart from this theory. It was found in these experiments that, bar thermal expansion effects, the spin-spin relaxation time is independent of temperature. The seventh chapter deals with the solid state and thermal mixing effects. The relevant theory, results obtained and a discussion of these results, are presented. The effect of various parameters on the polarisation rates and the 13C signal enhancement are investigated. Finally the effect of applying the DNP treatment on the central and hyperfine lines is discussed. The pulsed DNP process is presented in the eighth chapter. The relevant theory, the

  3. A new polarisation amplitude bias reduction method.

    NASA Astrophysics Data System (ADS)

    Vidal, M.; Leahy, J. P.; Dickinson, C.

    2016-06-01

    Polarisation amplitude estimation is affected by a positive noise bias, particularly important in regions with low signal-to-noise ratio (SNR). We present a new approach to correct for this bias in the case there is additional information about the polarisation angle. We develop the `known-angle estimator' that works in the special case when there is an independent and high signal-to-noise ratio (≳ 2σ) measurement of the polarisation angle. It is derived for the general case where the uncertainties in the Q, U Stokes parameters are not symmetric. This estimator completely corrects for the polarisation bias if the polarisation angle is perfectly known. In the realistic case, where the angle template has uncertainties, a small residual bias remains, but that is shown to be much smaller that the one left by other classical estimators. We also test our method with more realistic data, using the noise properties of the three lower frequency maps of WMAP. In this case, the known-angle estimator also produces better results than methods that do not include the angle information. This estimator is therefore useful in the case where the polarisation angle is expected to be constant over different data sets with different SNR.

  4. Computation of the internal forces in cilia: application to ciliary motion, the effects of viscosity, and cilia interactions.

    PubMed

    Gueron, S; Levit-Gurevich, K

    1998-04-01

    This paper presents a simple and reasonable method for generating a phenomenological model of the internal mechanism of cilia. The model uses a relatively small number of parameters whose values can be obtained by fitting to ciliary beat shapes. Here, we use beat patterns observed in Paramecium. The forces that generate these beats are computed and fit to a simple functional form called the "engine." This engine is incorporated into a recently developed hydrodynamic model that accounts for interactions between neighboring cilia and between the cilia and the surface from which they emerge. The model results are compared to data on ciliary beat patterns of Paramecium obtained under conditions where the beats are two-dimensional. Many essential features of the motion, including several properties that are not built in explicitly, are shown to be captured. In particular, the model displays a realistic change in beat pattern and frequency in response to increased viscosity and to the presence of neighboring cilia in configurations such as rows of cilia and two-dimensional arrays of cilia. We found that when two adjacent model cilia start beating at different phases they become synchronized within several beat periods, as observed in experiments where two flagella are brought into close proximity. Furthermore, examination of various multiciliary configurations shows that an approximately antiplectic wave pattern evolves autonomously. This modeling evidence supports earlier conjectures that metachronism may occur, at least partially, as a self-organized phenomenon due to hydrodynamic interactions between neighboring cilia. PMID:9545031

  5. Cilia-Associated Genes Play Differing Roles in Aminoglycoside-Induced Hair Cell Death in Zebrafish

    PubMed Central

    Stawicki, Tamara M.; Hernandez, Liana; Esterberg, Robert; Linbo, Tor; Owens, Kelly N.; Shah, Arish N.; Thapa, Nihal; Roberts, Brock; Moens, Cecilia B.; Rubel, Edwin W.; Raible, David W.

    2016-01-01

    Hair cells possess a single primary cilium, called the kinocilium, early in development. While the kinocilium is lost in auditory hair cells of most species it is maintained in vestibular hair cells. It has generally been believed that the primary role of the kinocilium and cilia-associated genes in hair cells is in the establishment of the polarity of actin-based stereocilia, the hair cell mechanotransduction apparatus. Through genetic screening and testing of candidate genes in zebrafish (Danio rerio) we have found that mutations in multiple cilia genes implicated in intraflagellar transport (dync2h1, wdr35, ift88, and traf3ip), and the ciliary transition zone (cc2d2a, mks1, and cep290) lead to resistance to aminoglycoside-induced hair cell death. These genes appear to have differing roles in hair cells, as mutations in intraflagellar transport genes, but not transition zone genes, lead to defects in kinocilia formation and processes dependent upon hair cell mechanotransduction activity. These mutants highlight a novel role of cilia-associated genes in hair cells, and provide powerful tools for further study. PMID:27207957

  6. Cilia-Associated Genes Play Differing Roles in Aminoglycoside-Induced Hair Cell Death in Zebrafish.

    PubMed

    Stawicki, Tamara M; Hernandez, Liana; Esterberg, Robert; Linbo, Tor; Owens, Kelly N; Shah, Arish N; Thapa, Nihal; Roberts, Brock; Moens, Cecilia B; Rubel, Edwin W; Raible, David W

    2016-01-01

    Hair cells possess a single primary cilium, called the kinocilium, early in development. While the kinocilium is lost in auditory hair cells of most species it is maintained in vestibular hair cells. It has generally been believed that the primary role of the kinocilium and cilia-associated genes in hair cells is in the establishment of the polarity of actin-based stereocilia, the hair cell mechanotransduction apparatus. Through genetic screening and testing of candidate genes in zebrafish (Danio rerio) we have found that mutations in multiple cilia genes implicated in intraflagellar transport (dync2h1, wdr35, ift88, and traf3ip), and the ciliary transition zone (cc2d2a, mks1, and cep290) lead to resistance to aminoglycoside-induced hair cell death. These genes appear to have differing roles in hair cells, as mutations in intraflagellar transport genes, but not transition zone genes, lead to defects in kinocilia formation and processes dependent upon hair cell mechanotransduction activity. These mutants highlight a novel role of cilia-associated genes in hair cells, and provide powerful tools for further study. PMID:27207957

  7. An age of enlightenment for cilia: The FASEB summer research conference on the "Biology of Cilia and Flagella".

    PubMed

    Tran, Pamela V; Lechtreck, Karl F

    2016-01-15

    From July 19-24, 2015, 169 clinicians and basic scientists gathered in the vertiginous heights of Snowmass, Colorado (2502 m) for the fourth FASEB summer research conference on the 'Biology of Cilia and Flagella'. Organizers Maureen Barr (Rutgers University), Iain Drummond (Massachusetts General Hospital/Harvard Medical School), and Jagesh Shah (Brigham and Women's Hospital/Harvard Medical School) assembled a program filled with new data and forward-thinking ideas documenting the ongoing growth of the field. Sixty oral presentations and 77 posters covered novel aspects of cilia structure, ciliogenesis, cilia motility, cilia-mediated signaling, and cilia-related disease. In this report, we summarize the meeting, highlight exciting developments and discuss open questions. PMID:26597000

  8. Structural defects in cilia of the choroid plexus, subfornical organ and ventricular ependyma are associated with ventriculomegaly

    PubMed Central

    2012-01-01

    Background Hydrocephalus is a heterogeneous disorder with multiple etiologies that are not yet fully understood. Animal models have implicated dysfunctional cilia of the ependyma and choroid plexus in the development of the disorder. In this report, we sought to determine the origin of the ventriculomegaly in four Bardet Biedl syndrome (BBS) mutant mouse strains as models of a ciliopathy. Methods Evans Blue dye was injected into the lateral ventricle of wild- type and BBS mutant mice to determine whether obstruction of intra- or extra-ventricular CSF flow contributed to ventriculomegaly. Transmission electron microscopy (TEM) was used to examine the ultrastructure of the choroid plexus, subfornical organ (SFO), subcommisural organ (SCO), and ventricular ependyma to evaluate their ultrastructure and the morphology of their primary and motile cilia. Results and discussion No obstruction of intra- or extra-ventricular CSF flow was observed, implying a communicating form of hydrocephalus in BBS mutant mice. TEM analyses of the mutants showed no evidence of choroidal papillomas or breakdown of the blood:CSF barrier. In contrast, structural defects were observed in a subpopulation of cilia lining the choroid plexus, SFO, and ventricular ependyma. These included disruptions of the microtubular structure of the axoneme and the presence of electron-dense vesicular-like material along the ciliary shaft and at the tips of cilia. Conclusions Abnormalities in cilia structure and function have the potential to influence ciliary intraflagellar transport (IFT), cilia maintenance, protein trafficking, and regulation of CSF production. Ciliary structural defects are the only consistent pathological features associated with CSF-related structures in BBS mutant mice. These defects are observed from an early age, and may contribute to the underlying pathophysiology of ventriculomegaly. PMID:23046663

  9. Biomimetic cilia arrays generate simultaneous pumping and mixing regimes

    PubMed Central

    Shields, A. R.; Fiser, B. L.; Evans, B. A.; Falvo, M. R.; Washburn, S.; Superfine, R.

    2010-01-01

    Living systems employ cilia to control and to sense the flow of fluids for many purposes, such as pumping, locomotion, feeding, and tissue morphogenesis. Beyond their use in biology, functional arrays of artificial cilia have been envisaged as a potential biomimetic strategy for inducing fluid flow and mixing in lab-on-a-chip devices. Here we report on fluid transport produced by magnetically actuated arrays of biomimetic cilia whose size approaches that of their biological counterparts, a scale at which advection and diffusion compete to determine mass transport. Our biomimetic cilia recreate the beat shape of embryonic nodal cilia, simultaneously generating two sharply segregated regimes of fluid flow: Above the cilia tips their motion causes directed, long-range fluid transport, whereas below the tips we show that the cilia beat generates an enhanced diffusivity capable of producing increased mixing rates. These two distinct types of flow occur simultaneously and are separated in space by less than 5 μm, approximately 20% of the biomimetic cilium length. While this suggests that our system may have applications as a versatile microfluidics device, we also focus on the biological implications of our findings. Our statistical analysis of particle transport identifying an enhanced diffusion regime provides novel evidence for the existence of mixing in ciliated systems, and we demonstrate that the directed transport regime is Poiseuille–Couette flow, the first analytical model consistent with biological measurements of fluid flow in the embryonic node. PMID:20798342

  10. Fetus Sound Stimulation: Cilia Memristor Effect of Signal Transduction

    PubMed Central

    Jankovic-Raznatovic, Svetlana; Dragojevic-Dikic, Svetlana; Rakic, Snezana; Nikolic, Branka; Plesinac, Snezana; Tasic, Lidija; Perisic, Zivko; Sovilj, Mirjana; Adamovic, Tatjana; Koruga, Djuro

    2014-01-01

    Background. This experimental study evaluates fetal middle cerebral artery (MCA) circulation after the defined prenatal acoustical stimulation (PAS) and the role of cilia in hearing and memory and could explain signal transduction and memory according to cilia optical-acoustical properties. Methods. PAS was performed twice on 119 no-risk term pregnancies. We analyzed fetal MCA circulation before, after first and second PAS. Results. Analysis of the Pulsatility index basic (PIB) and before PAS and Pulsatility index reactive after the first PAS (PIR 1) shows high statistical difference, representing high influence on the brain circulation. Analysis of PIB and Pulsatility index reactive after the second PAS (PIR 2) shows no statistical difference. Cilia as nanoscale structure possess magnetic flux linkage that depends on the amount of charge that has passed between two-terminal variable resistors of cilia. Microtubule resistance, as a function of the current through and voltage across the structure, leads to appearance of cilia memory with the “memristor” property. Conclusion. Acoustical and optical cilia properties play crucial role in hearing and memory processes. We suggest that fetuses are getting used to sound, developing a kind of memory patterns, considering acoustical and electromagnetically waves and involving cilia and microtubules and try to explain signal transduction. PMID:24719851

  11. New Measurements of Polarised Light from Exoplanets

    NASA Astrophysics Data System (ADS)

    Bott, Kimberly Marie; Cotton, Daniel; Kedziora-Chudczer, Lucyna; Bailey, Jeremy

    2015-08-01

    Detections of polarised light from exoplanets are an important expansion of exoplanet studies, as they provide a complimentary and advantageous diagnostic to the other characterisation methods. However some of the earliest claimed detections of polarised light from exoplanets are disputed.The High Precision Polarimetric Instrument (HIPPI) is currently the highest sensitivity astronomical polarimeter (Bailey 2015) in the world at 4.3 ppm or better precision, and has been used to observe exoplanet systems including the disputed first detection source (Berdyugina 2008, Wiktorowicz 2009, Berdyugina 2011), HD189733b. HIPPI is designed for best sensitivity in blue light where Rayleigh scattering would produce a strong signal for hot Jupiter planets (Burrows 2008). These observations, taken at the Anglo-Australian Telescope in 2014 and 2015, are compared to models from a polarised light adaptation of the Versatile Software for the Transfer of Atmospheric Radiation (VSTAR).Our new, independent Hot Jupiter polarised light detections are discussed, including the physical sources of the detected polarised light, as well as additional contributing factors (e.g. debris, ISM anomalies). For HD 189733b we use available visible and near infrared transit and polarimetric data to create and interpret a planetary portrait using VSTAR.

  12. Cilia-associated bacteria in fatal Bordetella bronchiseptica pneumonia of dogs and cats.

    PubMed

    Taha-Abdelaziz, Khaled; Bassel, Laura L; Harness, Melanie L; Clark, Mary Ellen; Register, Karen B; Caswell, Jeff L

    2016-07-01

    Bordetella bronchiseptica frequently causes nonfatal tracheobronchitis, but its role in fatal pneumonia is less recognized. Our study evaluated histologic identification of cilia-associated bacteria as a method for diagnosis of B. bronchiseptica pneumonia. Cases of fatal bronchopneumonia were studied retrospectively, excluding neonates and cases of aspiration pneumonia, minor lung lesions, or autolysis. The study population comprised 36 canine and 31 feline cases of bronchopneumonia. B. bronchiseptica was identified in 8 of 36 canine and 14 of 31 feline cases based on immunohistochemistry (IHC) using serum from a rabbit hyperimmunized with pertactin, PCR testing (Fla2/Fla12), and/or bacterial culture data when available. Of these, IHC was positive in 4 canine and 7 feline cases, PCR was positive in 8 canine and 14 feline cases, and B. bronchiseptica was isolated in 2 of 5 canine and 3 of 9 feline cases tested. Examination of histologic sections stained with hematoxylin and eosin revealed bronchial cilia-associated bacteria in 4 of 36 canine and 5 of 31 feline cases; these were all positive by IHC and PCR. The presence of cilia-associated bacteria had been noted in the pathology report for only 2 of these 9 cases. Thus, the presence of cilia-associated bacteria seems frequently overlooked by pathologists, but is a diagnostically significant feature of B. bronchiseptica pneumonia. A specific diagnosis of B. bronchiseptica pneumonia is important because it suggests primary or opportunistic bacterial pneumonia rather than aspiration pneumonia, and because of the risk of animal-to-animal transmission of B. bronchiseptica, the availability of vaccines for disease prevention, and the potential zoonotic risk to immunocompromised pet owners. PMID:27178716

  13. Linking the Primary Cilium to Cell Migration in Tissue Repair and Brain Development

    PubMed Central

    Veland, Iben Rønn; Lindbæk, Louise; Christensen, Søren Tvorup

    2014-01-01

    Primary cilia are unique sensory organelles that coordinate cellular signaling networks in vertebrates. Inevitably, defects in the formation or function of primary cilia lead to imbalanced regulation of cellular processes that causes multisystemic disorders and diseases, commonly known as ciliopathies. Mounting evidence has demonstrated that primary cilia coordinate multiple activities that are required for cell migration, which, when they are aberrantly regulated, lead to defects in organogenesis and tissue repair, as well as metastasis of tumors. Here, we present an overview on how primary cilia may contribute to the regulation of the cellular signaling pathways that control cyclic processes in directional cell migration. PMID:26955067

  14. The significance of ultrastructural abnormalities of human cilia.

    PubMed

    Fox, B; Bull, T B; Makey, A R; Rawbone, R

    1981-12-01

    The electronmicroscopic structure of cilia was studied from the inferior turbinate of the nose in 22 adults, and in 84 biopsies from the bronchial tree of 40 adults. The incidence of compound cilia and abnormal microtubular structures was assessed. There were significant variations in the incidence of abnormalities in different parts of the airways and even within different areas of the same electronmicroscopic section. The focal nature of differences in structure of cilia indicate that abnormalities found in a single biopsy do not necessarily reflect a generalized change in the bronchial tree. Thus, such a finding should not be used as evidence that the abnormalities of cilia are the cause of decrease in mucociliary clearance or that they play a role in the pathogenesis of bronchiectasis and sinusitis. PMID:7307613

  15. Hydrodynamic interactions of cilia on a spherical body

    NASA Astrophysics Data System (ADS)

    Nasouri, Babak; Elfring, Gwynn J.

    2015-11-01

    The emergence of metachronal waves in ciliated microorganisms can arise solely from the hydrodynamic interactions between the cilia. For a chain of cilia attached to a flat ciliate, it was observed that fluid forces can lead the system to form a metachronal wave. However, several microorganisms such as paramecium and volvox possess a curved shaped ciliate body. To understand the effect of this geometry on the formation of metachronal waves, we evaluate the hydrodynamic interactions of cilia near a large spherical body. Using a minimal model, we show that for a chain of cilia around the sphere, the embedded periodicity in the geometry leads the system to synchronize. We also report an emergent wave-like behavior when an asymmetry is introduced to the system.

  16. Using Xenopus Skin to Study Cilia Development and Function

    PubMed Central

    Werner, Michael E.; Mitchell, Brian J.

    2015-01-01

    Cilia are prevalent biological structures that are important for cell signaling and for generating fluid flow (or motility). Cilia are found throughout biology from single-celled organisms to vertebrates, and many model systems have been employed for their analysis. Here, we describe the use of Xenopus larval skin as a system for the study of ciliogenesis and ciliary function. In particular, we describe basic molecular and embryological manipulations and imaging techniques that have proven particularly useful for understanding the polarized beating of cilia and the generation of directed fluid flow (Werner & Mitchell, 2012). However, these same tools have the potential to benefit a large number of cilia-related biological questions. PMID:23522471

  17. Microscale imaging of cilia-driven fluid flow

    PubMed Central

    Huang, Brendan K.; Choma, Michael A.

    2015-01-01

    Cilia-driven fluid flow is important for multiple processes in the body, including respiratory mucus clearance, gamete transport in the oviduct, right-left patterning in the embryonic node, and cerebrospinal fluid circulation. Multiple imaging techniques have been applied towards quantifying ciliary flow. Here we review common velocimetry methods of quantifying fluid flow. We then discuss four important optical modalities, including light microscopy, epifluorescence, confocal microscopy, and optical coherence tomography, that have been used to investigate cilia-driven flow. PMID:25417211

  18. Endothelial Cilia Are Essential for Developmental Vascular Integrity in Zebrafish

    PubMed Central

    Kallakuri, Sowjanya; Yu, Jianxin A.; Li, Jade; Li, Yuanyuan; Weinstein, Brant M.; Nicoli, Stefania

    2015-01-01

    The cilium is a signaling platform of the vertebrate cell. It has a critical role in polycystic kidney disease and nephronophthisis. Cilia have been detected on endothelial cells, but the function of these organelles in the vasculature remains incompletely defined. In this study, using genetic and chemical genetic tools in the model organism zebrafish, we reveal an essential role of cilia in developmental vascular integrity. Embryos expressing mutant intraflagellar transport genes, which are essential and specific for cilia biogenesis, displayed increased risk of developmental intracranial hemorrhage, whereas the morphology of the vasculature remained normal. Moreover, cilia were present on endothelial cells in the developing zebrafish vasculature. We further show that the involvement of cilia in vascular integrity is endothelial autonomous, because endothelial-specific re-expression of intraflagellar transport genes in respective mutants rescued the intracranial hemorrhage phenotype. Finally, whereas inhibition of Hedgehog signaling increased the risk of intracranial hemorrhage in ciliary mutants, activation of the pathway rescued this phenotype. In contrast, embryos expressing an inactivating mutation in pkd2, one of two autosomal dominant cystic kidney disease genes, did not show increased risk of developmental intracranial hemorrhage. These results suggest that Hedgehog signaling is a major mechanism for this novel role of endothelial cilia in establishing vascular integrity. PMID:25214579

  19. Pneumatically-actuated artificial cilia array for biomimetic fluid propulsion.

    PubMed

    Gorissen, Benjamin; de Volder, Michaël; Reynaerts, Dominiek

    2015-11-21

    Arrays of beating cilia emerged in nature as one of the most efficient propulsion mechanisms at a small scale, and are omnipresent in microorganisms. Previous attempts at mimicking these systems have foundered against the complexity of fabricating small-scale cilia exhibiting complex beating motions. In this paper, we propose for the first time arrays of pneumatically-actuated artificial cilia that are able to address some of these issues. These artificial cilia arrays consist of six highly flexible silicone rubber actuators with a diameter of 1 mm and a length of 8 mm that can be actuated independently from each other. In an experimental setup, the effects of the driving frequency, phase difference and duty cycle on the net flow in a closed-loop channel have been studied. Net fluid speeds of up to 19 mm s(-1) have been measured. Further, it is possible to invert the flow direction by simply changing the driving frequency or by changing the duty cycle of the driving block pulse pressure wave without changing the bending direction of the cilia. Using PIV measurements, we corroborate for the first time existing mathematical models of cilia arrays to measurements on prototypes. PMID:26439855

  20. Asynchronous beating of cilia enhances particle capture rate

    NASA Astrophysics Data System (ADS)

    Ding, Yang; Kanso, Eva

    2014-11-01

    Many aquatic micro-organisms use beating cilia to generate feeding currents and capture particles in surrounding fluids. One of the capture strategies is to ``catch up'' with particles when a cilium is beating towards the overall flow direction (effective stroke) and intercept particles on the downstream side of the cilium. Here, we developed a 3D computational model of a cilia band with prescribed motion in a viscous fluid and calculated the trajectories of the particles with different sizes in the fluid. We found an optimal particle diameter that maximizes the capture rate. The flow field and particle motion indicate that the low capture rate of smaller particles is due to the laminar flow in the neighbor of the cilia, whereas larger particles have to move above the cilia tips to get advected downstream which decreases their capture rate. We then analyzed the effect of beating coordination between neighboring cilia on the capture rate. Interestingly, we found that asynchrony of the beating of the cilia can enhance the relative motion between a cilium and the particles near it and hence increase the capture rate.

  1. Generating conditional mutants to analyze ciliary functions: the use of Cre-lox technology to disrupt cilia in specific organs.

    PubMed

    O'Connor, Amber K; Kesterson, Robert A; Yoder, Bradley K

    2009-01-01

    The list of human disordered associated with cilia dysfunction, the ciliopathies, continues to highlight the importance of understanding the many roles of the long overlooked primary cilium. Much of the insights into the clinical importance of the cilium have come from analyses in model organisms, especially the mouse. However, the early embryonic lethality and severe developmental defects associated with cilia disruption has hindered progress in exploring cilia functions in late development or in adult tissues. This hurdle is being surmounted through the use of conditional alleles of genes encoding ciliary proteins and Cre deletor lines with inducible Cre activity or with lines expressing Cre in a cell-type-specific manner. Results from these approaches are providing important insights into the diverse array of cellular and tissue activities regulated by the cilium. Here we provide a recent account of the Cre/lox strategy. The generation and use of well-designed conditional alleles, as well as careful manipulation of embryonic stem cells are discussed. We also provide specific examples to illustrate the use of Cre/lox approaches to evaluate ciliary function in several tissues. With the recent characterization of multiple cilia proteomes along with efforts of several consortia to generate conditional alleles of all genes in the mouse, further use of conditional mutation approaches promise to yield many advances and surprises as we explore the functions of this increasingly complex organelle. PMID:20409823

  2. A high-resolution morphological and ultrastructural map of anterior sensory cilia and glia in Caenorhabditis elegans

    PubMed Central

    Doroquez, David B; Berciu, Cristina; Anderson, James R; Sengupta, Piali; Nicastro, Daniela

    2014-01-01

    Many primary sensory cilia exhibit unique architectures that are critical for transduction of specific sensory stimuli. Although basic ciliogenic mechanisms are well described, how complex ciliary structures are generated remains unclear. Seminal work performed several decades ago provided an initial but incomplete description of diverse sensory cilia morphologies in C. elegans. To begin to explore the mechanisms that generate these remarkably complex structures, we have taken advantage of advances in electron microscopy and tomography, and reconstructed three-dimensional structures of fifty of sixty sensory cilia in the C. elegans adult hermaphrodite at high resolution. We characterize novel axonemal microtubule organization patterns, clarify structural features at the ciliary base, describe new aspects of cilia–glia interactions, and identify structures suggesting novel mechanisms of ciliary protein trafficking. This complete ultrastructural description of diverse cilia in C. elegans provides the foundation for investigations into underlying ciliogenic pathways, as well as contributions of defined ciliary structures to specific neuronal functions. DOI: http://dx.doi.org/10.7554/eLife.01948.001 PMID:24668170

  3. 9 + 0 and 9 + 2 cilia are randomly dispersed in the mouse node.

    PubMed

    Odate, Toru; Takeda, Sen; Narita, Keishi; Kawahara, Toru

    2016-04-01

    The initial determination of left-right asymmetry is an essential process in embryonic development. In mouse embryo, cilia in the node play an important role generating the nodal flow that subsequently triggers left-right determination in the embryo. Although nodal cilia have historically been thought to have a 9 + 0 axonemal configuration, the existence of 9 + 2 cilia has been reported so far. Because the distribution of those two types of cilia within the node has not yet been reported, we assessed the arrangement of 9 + 0 and 9 + 2 cilia in the node. In this study, we concluded that most of the nodal cilia were 9 + 0 in structure and there were much fewer 9 + 2 cilia than 9 + 0 cilia. Furthermore, the two types of cilia were randomly distributed in the node with no regularity. In addition, we studied the embryonic origin of the crown cells surrounding the node to better understand their identity. PMID:26520785

  4. Polarisation modulation in X-ray binaries

    NASA Astrophysics Data System (ADS)

    Ingram, Adam; Maccarone, Thomas

    2016-07-01

    X-ray polarimetry promises to provide a powerful new lever arm for studying accretion onto black holes with the next generation of X-ray telescopes. I will discuss how polarisation can be used to help constrain the physical origin of quasi-periodic oscillations (QPOs) observed in the X-ray light curves of accreting black holes. QPOs may be signatures of the frame dragging effect: in General Relativity, a spinning black hole twists up the surrounding space-time, causing vertical precession of nearby orbits. In the truncated disc / precessing inner flow model, the entire inner accretion flow precesses as a solid body causing a modulation in the X-ray flux through solid angle and Doppler effects. This model also predicts the observed polarisation of the X-ray signal to vary quasi-periodically. I will summarise our work to model the polarisation signal from a precessing accretion flow, starting with simple assumptions about the emission mechanism but taking General Relativity fully into account. We find that it should be possible to measure the predicted modulation in polarisation degree for a reasonable region of parameter space with a polarimeter capable of detecting ~60 counts per second from a bright black hole binary. I will also show that sensitivity can be greatly improved by correlating the signal with a high count rate reference band signal.

  5. Optimal polarisation equations in FLRW universes

    SciTech Connect

    Tram, Thomas; Lesgourgues, Julien E-mail: Julien.Lesgourgues@cern.ch

    2013-10-01

    This paper presents the linearised Boltzmann equation for photons for scalar, vector and tensor perturbations in flat, open and closed FLRW cosmologies. We show that E- and B-mode polarisation for all types can be computed using only a single hierarchy. This was previously shown explicitly for tensor modes in flat cosmologies but not for vectors, and not for non-flat cosmologies.

  6. Midbody remnant licenses primary cilia formation in epithelial cells.

    PubMed

    Ott, Carolyn M

    2016-08-01

    Tethered midbody remnants dancing across apical microvilli, encountering the centrosome, and beckoning forth a cilium-who would have guessed this is how polarized epithelial cells coordinate the end of mitosis and the beginning of ciliogenesis? New evidence from Bernabé-Rubio et al. (2016. J. Cell Biol http://dx.doi.org/10.1083/jcb.201601020) supports this emerging model. PMID:27482049

  7. The coiled-coil domain containing protein CCDC151 is required for the function of IFT-dependent motile cilia in animals.

    PubMed

    Jerber, Julie; Baas, Dominique; Soulavie, Fabien; Chhin, Brigitte; Cortier, Elisabeth; Vesque, Christine; Thomas, Joëlle; Durand, Bénédicte

    2014-02-01

    Cilia are evolutionarily conserved organelles endowed with essential physiological and developmental functions. In humans, disruption of cilia motility or signaling leads to complex pleiotropic genetic disorders called ciliopathies. Cilia motility requires the assembly of multi-subunit motile components such as dynein arms, but mechanisms underlying their assembly pathway and transport into the axoneme are still largely unknown. We identified a previously uncharacterized coiled-coil domain containing protein CCDC151, which is evolutionarily conserved in motile ciliated species and shares ancient features with the outer dynein arm-docking complex 2 of Chlamydomonas. In Drosophila, we show that CG14127/CCDC151 is associated with motile intraflagellar transport (IFT)-dependent cilia and required for geotaxis behavior of adult flies. In zebrafish, Ccdc151 is expressed in tissues with motile cilia, and morpholino-induced depletion of Ccdc151 leads to left-right asymmetry defects and kidney cysts. We demonstrate that Ccdc151 is required for proper motile function of cilia in the Kupffer's vesicle and in the pronephros by controlling dynein arm assembly, showing that Ccdc151 is a novel player in the control of IFT-dependent dynein arm assembly in animals. However, we observed that CCDC151 is also implicated in other cellular functions in vertebrates. In zebrafish, ccdc151 is involved in proper orientation of cell divisions in the pronephros and genetically interacts with prickle1 in this process. Furthermore, knockdown experiments in mammalian cells demonstrate that CCDC151 is implicated in the regulation of primary cilium length. Hence, CCDC151 is required for motile cilia function in animals but has acquired additional non-motile functions in vertebrates. PMID:24067530

  8. Oscillations of Eukaryotic Cilia and Flagella

    NASA Astrophysics Data System (ADS)

    Gopinath, Arvind; Mahadevan, Lakshminarayanan

    2006-11-01

    The undulating beat of eukaryotic flagella and cilia produces forces that move cells and cause locomotion. The timing mechanisms that generate these periodic undulations are still mysterious and the question of how these oscillations arise is still a subject of much research - both experimental and theoretical. Recent experimental results on paralyzed and reconstituted flagella offer new insight into the dynamical mechanisms that could result in sustained waveform generation. Motivated by these recent experimental results we propose a model that mimics the flagellar structure as motor driven elastic, inextensible filaments. We hypothesize that the oscillations arise due to motor (dynein) driven, constrained, relative sliding of parts of the flagella. The dynamical equations describing the evolution of the populations of attached and detached motors is actively coupled to the local configuration as well as local sliding velocities via strain and configuration dependent kinetic reaction rates. At the same time, the filament configuration is actively coupled to the motor densities via the dependence of the active internal torque densities on the motor populations as well as their internal state. Appropriate ensemble averaged force-velocity relationships for the motors completes the set of equations. Numerical solutions reveal onset of dynamical instabilities via Hopf-bifurcations with oscillatory waveforms emerging from a trivial base state corresponding to a straight, non-moving flagellum.

  9. Caught Nek-ing: cilia and centrioles.

    PubMed

    Quarmby, Lynne M; Mahjoub, Moe R

    2005-11-15

    The Nek family of cell-cycle kinases is widely represented in eukaryotes and includes numerous proteins that were described only recently and remain poorly characterized. Comparing Neks in the context of clades allows us to examine the question of whether microbial eukaryotic Neks, although not strictly orthologs of their vertebrate counterparts, can provide clues to ancestral functions that might be retained in the vertebrate Neks. Relatives of the Nek2/NIMA proteins play important roles at the G2-M transition in nuclear envelope breakdown and centromere separation. Nek6, Nek7 and Nek9 also seem to regulate mitosis. By contrast, Nek1 and Nek8 have been linked with polycystic kidney disease. Results of statistical analysis indicate that the family coevolved with centrioles that function as both microtubule-organizing centers and the basal bodies of cilia. This evolutionary perspective, taken together with functional studies of microbial Neks, provides new insights into the cellular roles of the proteins and disease with which some of them have been linked. PMID:16280549

  10. Polarisation of electroweak gauge bosons at the LHC

    NASA Astrophysics Data System (ADS)

    Stirling, James; Vryonidou, Eleni

    2013-05-01

    We present results for the polarisation of gauge bosons produced at the LHC. Polarisation effects for W bosons manifest themselves in the angular distributions of the lepton and in the distributions of lepton transverse momentum and missing transverse energy. The polarisation is discussed for a range of different processes producing W bosons such as W+jets and W from top production. The relative contributions of the different polarisation states vary from process to process, reflecting the dynamics of the underlying hardscattering process. We also calculate the polarisation of the Z boson produced in association with QCD jets at the LHC.

  11. Experimental investigation of the flow induced by artificial cilia.

    PubMed

    Hussong, J; Schorr, N; Belardi, J; Prucker, O; Rühe, J; Westerweel, J

    2011-06-21

    The fluid transport produced by rectangular shaped, magnetically actuated artificial cilia of 70 μm length and 20 μm width was determined by means of phase-locked Micro Particle Image Velocimetry (μPIV) measurements in a closed microfluidic chamber. The phase-averaged flow produced by the artificial cilia reached up to 130 μm s(-1) with an actuation cycle frequency of 10 Hz. Analysis of the measured flow data indicate that the present system is capable of achieving volume flow rates of V[combining dot above](cilia) = 14 ± 4 μl min(-1) in a micro channel of 0.5 × 5 mm(2) cross-sectional area when no back pressure is built up. This corresponds to an effective pressure gradient of 6 ± 1 Pa m(-1), which equals a pressure difference of 0.6 ± 0.1 mPa over a distance of 100 μm between two rows of cilia. These results were derived analytically from the measured velocity profile by treating the cilia as a thin boundary layer. While the cilia produce phase-averaged velocities of the order of O(10(2)μm s(-1)), time-resolved measurements showed that the flow field reverses two times during one actuation cycle inducing instantaneous velocities of up to approximately 2 mm s(-1). This shows that the flow field is dominated by fluid oscillations and flow rates are expected to increase if the beating motion of the cilia is further improved. PMID:21614349

  12. Effect of viscosity on metachrony in mucus propelling cilia.

    PubMed

    Gheber, L; Korngreen, A; Priel, Z

    1998-01-01

    In the present work we report that increasing the viscosity of the medium caused not only a decrease in the ciliary beat frequency but also changes in the metachrony and correlation between cilia. The study was performed using double and triple simultaneous photoelectric measurements on cultured ciliary cells from the frog esophagus in the viscosity range of 1-2,000 cp. We observed that increasing the viscosity intensified the fluctuations in all the measured parameters. Ciliary beat frequency decreased moderately. Even at quite high viscosities (circa 2000 cp.), cilia were still active with beating frequencies of 3-5 Hz. In addition, the degree of correlation between cilia parallel to the effective stroke direction (ESD) decreased, while that perpendicular to the ESD at a low range of viscosities remained unchanged and even increased at high viscosities. Medium viscosities in the range of 30-1,500 cp. altered the metachronal wave properties of cultured frog esophagus. The metachronal wavelength increased by up to 50%, and the wave direction changed towards more orthoplectic type of coordination. According to our recently suggested model [Gheber and Priel, 1990: Cell Motil. Cytoskeleton 16:167-181], these effects can be explained by a decrease in the temporal asymmetry of the ciliary beat. Since similar results were observed in water propelling cilia of Paramecium subjected to medium viscosity ranges of up to 40 cp. [Machemer, 1972: J. Exp. Biol. 57:239-259], we conclude that hydrodynamic interactions govern the metachronal wave properties of both mucus and water propelling cilia, though mucus propelling cilia, with their better adaptation to increased load, are affected at much higher viscosities than water propelling cilia. PMID:9453710

  13. A Numerical Study of Muco-Ciliary Transport under the condition of Primary Ciliary Dyskinesia

    NASA Astrophysics Data System (ADS)

    Jayathilake, Pahala Gedara; Lee, Wan Lung; Le, Duc Vinh; Lee, Heow Pueh; Khoo, Boo Cheong

    2012-11-01

    Primary ciliary dyskinesia (PCD) is a disease due to the defects in motile cilia. A two-dimensional numerical model based on the immersed boundary method coupled with the projection method is used for a preliminary study of the flow physics of muco-ciliary transport of human respiratory tract under PCD conditions. The effects of the cilia beating amplitude, cilia beat pattern (CBP), cilia beat frequency (CBF), immotile cilia, and uncoordinated beating of cilia on mucus transport are investigated. As expected, the mucus velocity decreases as the beating amplitude and CBF decrease. The windscreen wiper motion and rigid rod motion, which are two abnormal CBPs owing to PCD, would greatly reduce the mucus transport. The mucus velocity decreases rather linearly if the number of uniformly distributed immotile cilia increases. The results further show that the mucus velocity would be slightly reduced when the uniformly distributed immotile cilia are rearranged as a cluster of immotile cilia. Furthermore, if the half of the cilia are immotile and uniformly distributed, the incoordination between motile cilia would not significantly affect the mucus velocity.

  14. Microscale flow propulsion through bioinspired and magnetically actuated artificial cilia.

    PubMed

    Chen, Chia-Yuan; Cheng, Ling-Ying; Hsu, Chun-Chieh; Mani, Karthick

    2015-05-01

    Recent advances in microscale flow propulsion through bioinspired artificial cilia provide a promising alternative for lab-on-a-chip applications. However, the ability of actuating artificial cilia to achieve a time-dependent local flow control with high accuracy together with the elegance of full integration into the biocompatible microfluidic platforms remains remote. Driven by this motive, the current work has constructed a series of artificial cilia inside a microchannel to facilitate the time-dependent flow propulsion through artificial cilia actuation with high-speed (>40 Hz) circular beating behavior. The generated flow was quantified using micro-particle image velocimetry and particle tracking with instantaneous net flow velocity of up to 10(1 ) μm/s. Induced flow patterns caused by the tilted conical motion of artificial cilia constitutes efficient fluid propulsion at microscale. This flow phenomenon was further measured and illustrated by examining the induced flow behavior across the depth of the microchannel to provide a global view of the underlying flow propulsion mechanism. The presented analytic paradigms and substantial flow evidence present novel insights into the area of flow manipulation at microscale. PMID:26045730

  15. Microscale flow propulsion through bioinspired and magnetically actuated artificial cilia

    PubMed Central

    Chen, Chia-Yuan; Cheng, Ling-Ying; Hsu, Chun-Chieh; Mani, Karthick

    2015-01-01

    Recent advances in microscale flow propulsion through bioinspired artificial cilia provide a promising alternative for lab-on-a-chip applications. However, the ability of actuating artificial cilia to achieve a time-dependent local flow control with high accuracy together with the elegance of full integration into the biocompatible microfluidic platforms remains remote. Driven by this motive, the current work has constructed a series of artificial cilia inside a microchannel to facilitate the time-dependent flow propulsion through artificial cilia actuation with high-speed (>40 Hz) circular beating behavior. The generated flow was quantified using micro-particle image velocimetry and particle tracking with instantaneous net flow velocity of up to 101 μm/s. Induced flow patterns caused by the tilted conical motion of artificial cilia constitutes efficient fluid propulsion at microscale. This flow phenomenon was further measured and illustrated by examining the induced flow behavior across the depth of the microchannel to provide a global view of the underlying flow propulsion mechanism. The presented analytic paradigms and substantial flow evidence present novel insights into the area of flow manipulation at microscale. PMID:26045730

  16. Ciliopathy-associated gene Cc2d2a promotes assembly of subdistal appendages on the mother centriole during cilia biogenesis

    PubMed Central

    Veleri, Shobi; Manjunath, Souparnika H.; Fariss, Robert N.; May-Simera, Helen; Brooks, Matthew; Foskett, Trevor A.; Gao, Chun; Longo, Teresa A.; Liu, Pinghu; Nagashima, Kunio; Rachel, Rivka A.; Li, Tiansen; Dong, Lijin; Swaroop, Anand

    2014-01-01

    The primary cilium originates from the mother centriole and participates in critical functions during organogenesis. Defects in cilia biogenesis or function lead to pleiotropic phenotypes. Mutations in centrosome-cilia gene CC2D2A result in Meckel and Joubert syndromes. Here we generate a Cc2d2a-/- mouse that recapitulates features of Meckel syndrome including embryonic lethality and multi-organ defects. Cilia are absent in Cc2d2a-/- embryonic node and other somatic tissues; disruption of cilia-dependent Shh signaling appears to underlie exencephaly in mutant embryos. The Cc2d2a-/- mouse embryonic fibroblasts (MEFs) lack cilia though mother centriole and pericentriolar proteins are detected. Odf2, associated with subdistal appendages, is absent and ninein is reduced in mutant MEFs. In Cc2d2a-/- MEFs, subdistal appendages are lacking or abnormal by transmission-EM. Consistent with this, CC2D2A localizes to subdistal appendages by immuno-EM in wild type cells. We conclude that CC2D2A is essential for the assembly of subdistal appendages, which anchor cytoplasmic microtubules and prime the mother centriole for axoneme biogenesis. PMID:24947469

  17. Modelling elliptically polarised free electron lasers

    NASA Astrophysics Data System (ADS)

    Henderson, J. R.; Campbell, L. T.; Freund, H. P.; McNeil, B. W. J.

    2016-06-01

    A model of a free electron laser (FEL) operating with an elliptically polarised undulator is presented. The equations describing the FEL interaction, including resonant harmonic radiation fields, are averaged over an undulator period and generate a generalised Bessel function scaling factor, similar to that of planar undulator FEL theory. Comparison between simulations of the averaged model with those of an unaveraged model show very good agreement in the linear regime. Two unexpected results were found. Firstly, an increased coupling to harmonics for elliptical rather than planar polarisarised undulators. Secondly, and thought to be unrelated to the undulator polarisation, a significantly different evolution between the averaged and unaveraged simulations of the harmonic radiation evolution approaching FEL saturation.

  18. Centriole distal appendages promote membrane docking, leading to cilia initiation

    PubMed Central

    Tanos, Barbara E.; Yang, Hui-Ju; Soni, Rajesh; Wang, Won-Jing; Macaluso, Frank P.; Asara, John M.; Tsou, Meng-Fu Bryan

    2013-01-01

    The distal appendages (DAPs) of centrioles have been proposed to anchor cilia to the plasma membrane, but their molecular composition, assembly, and exact function in ciliogenesis remain poorly understood. Using quantitative centrosome proteomics and superresolution microscopy, we identified five DAP components, including one previously described (CEP164), one partially characterized (CEP89 [ccdc123]), and three novel (CEP83 [ccdc41], SCLT1, and FBF1) DAP proteins. Analyses of DAP assembly revealed a hierarchy. CEP83 recruits both SCLT1 and CEP89 to centrioles. Subsequent recruitment of FBF1 and CEP164 is independent of CEP89 but mediated by SCLT1. All five DAP components are essential for ciliogenesis; loss of CEP83 specifically blocks centriole-to-membrane docking. Undocked centrioles fail to recruit TTBK2 or release CP110, the two earliest modifications found on centrioles prior to cilia assembly, revealing centriole-to-membrane docking as a temporal and spatial cue promoting cilia initiation. PMID:23348840

  19. Novel Insights into the Development and Function of Cilia Using the Advantages of the Paramecium Cell and Its Many Cilia

    PubMed Central

    Yano, Junji; Valentine, Megan S.; Van Houten, Judith L.

    2015-01-01

    Paramecium species, especially P. tetraurelia and caudatum, are model organisms for modern research into the form and function of cilia. In this review, we focus on the ciliary ion channels and other transmembrane proteins that control the beat frequency and wave form of the cilium by controlling the signaling within the cilium. We put these discussions in the context of the advantages that Paramecium brings to the understanding of ciliary motility: mutants for genetic dissections of swimming behavior, electrophysiology, structural analysis, abundant cilia for biochemistry and modern proteomics, genomics and molecular biology. We review the connection between behavior and physiology, which allows the cells to broadcast the function of their ciliary channels in real time. We build a case for the important insights and advantages that this model organism continues to bring to the study of cilia. PMID:26230712

  20. Novel Insights into the Development and Function of Cilia Using the Advantages of the Paramecium Cell and Its Many Cilia.

    PubMed

    Yano, Junji; Valentine, Megan S; Van Houten, Judith L

    2015-01-01

    Paramecium species, especially P. tetraurelia and caudatum, are model organisms for modern research into the form and function of cilia. In this review, we focus on the ciliary ion channels and other transmembrane proteins that control the beat frequency and wave form of the cilium by controlling the signaling within the cilium. We put these discussions in the context of the advantages that Paramecium brings to the understanding of ciliary motility: mutants for genetic dissections of swimming behavior, electrophysiology, structural analysis, abundant cilia for biochemistry and modern proteomics, genomics and molecular biology. We review the connection between behavior and physiology, which allows the cells to broadcast the function of their ciliary channels in real time. We build a case for the important insights and advantages that this model organism continues to bring to the study of cilia. PMID:26230712

  1. Model Cilia - Experiments with Biomimetic Actuable Structures and Surfaces

    NASA Astrophysics Data System (ADS)

    Lloyd Carroll, R.

    2005-03-01

    The use of cilia to drive fluid flow is a common motif in living organisms, and in the tissues of higher organisms. By understanding the ways that cilia function (or do not function), potential therapies to treat human diseases (such as cystic fibrosis) may be devised. The complex hydrodynamics of flow in beating ciliary tissues (such as lung epithelial tissues) are challenging to study in cultured tissues, suggesting the need for model systems that will mimic the morphology and beat patterns of living systems. To reach this goal, we have fabricated high aspect ratio cilia-like structures with dimensions similar to those of a lung epithelial cilium (0.2 to 2.0 μm diameter by ˜6 to 10 μm long). The structures and surfaces are composed of a magneto-elastomeric nanocomposite, allowing the actuation of artificial cilia by magnetic fields. We have studied the flexibility of the materials under conditions of flow (in microfluidics channels), and will present theoretical and experimental data from various efforts at actuation. We will discuss details of the fabrication of the ciliated structures and present results of mechanical characterization. The impact of this work on the understanding of fluid flow above ciliated cells and tissues and potential applications of such model systems will also be described.

  2. Realizing the Physics of Motile Cilia Synchronization with Driven Colloids

    NASA Astrophysics Data System (ADS)

    Bruot, Nicolas; Cicuta, Pietro

    2016-03-01

    Cilia and flagella in biological systems often show large scale cooperative behaviors such as the synchronization of their beats in "metachronal waves." These are beautiful examples of emergent dynamics in biology, and are essential for life, allowing diverse processes from the motility of eukaryotic microorganisms, to nutrient transport and clearance of pathogens from mammalian airways. How these collective states arise is not fully understood, but it is clear that individual cilia interact mechanically, and that a strong and long-ranged component of the coupling is mediated by the viscous fluid. We review here the work by ourselves and others aimed at understanding the behavior of hydrodynamically coupled systems, and particularly a set of results that have been obtained both experimentally and theoretically by studying actively driven colloidal systems. In these controlled scenarios, it is possible to selectively test aspects of living motile cilia, such as the geometrical arrangement, the effects of the driving profile and the distance to no-slip boundaries. We outline and give examples of how it is possible to link model systems to observations on living systems, which can be made on microorganisms, on cell cultures or on tissue sections. This area of research has clear clinical application in the long term, as severe pathologies are associated with compromised cilia function in humans.

  3. Symmetry Breaking in a Model for Nodal Cilia

    NASA Astrophysics Data System (ADS)

    Brokaw, Charles J.

    2005-03-01

    Nodal cilia are very short cilia found in the embryonic node on the ventral surface of early mammalian embryos. They create a right to left fluid flow that is responsible for determining the normal asymmetry of the internal organs of the mammalian body. To do this, the distal end of the cilium must circle in a counterclockwise sense. Computer simulations with 3-dimensional models of flagella allow examination of 3-dimensional movements such as those of nodal cilia. 3-dimensional circling motions of short cilia can be achieved with velocity controlled models, in which dynein activity is regulated by sliding velocity. If dyneins on one outer doublet are controlled by the sliding velocity experienced by that doublet, the system is symmetric, and the 3-dimensional models can show either clockwise or counterclockwise circling. My computer simulations have examined two possible symmetry breaking mechanisms: 1) dyneins on doublet N are regulated by a mixture of the sliding velocities experienced by doublets N and N+1 (numbered in a clockwise direction, looking from the base). or 2) symmetry is broken by an off-axis force that produces a right-handed twist of the axoneme, consistent with observations that some dyneins can rotate their substrate microtubules in a clockwise direction.

  4. Magnetically actuated artificial cilia for optimum mixing performance in microfluidics.

    PubMed

    Chen, Chia-Yuan; Chen, Chia-Yun; Lin, Cheng-Yi; Hu, Ya-Ting

    2013-07-21

    Contemporary lab-chip devices require efficient, high-performance mixing capability. A series of artificial cilia with embedded magnetic particles was fabricated to achieve precise flow manipulation through magnetically driven control. These fabricated structures were actuated in a homogeneous magnetic field generated by a built-in magnetic coil system for various beating cycles inside a microchannel. Three representative trajectories, namely, circular motion, back-and-forth oscillation, and a figure-of-eight pattern, of artificial cilia were designed and generated to mimic the motion of actual cilia. Homogeneous mixing of two highly viscous (>25 centipoise) dyed solutions by using the figure-of-eight trajectory achieved a mixing efficiency of approximately 86%. The underlying relationship between ciliated structures and the induced flow fields was further elucidated by performing a hydrodynamic analysis with micro-particle image velocimetry. In addition, a numerical modeling method which used a fluid structure interaction module was applied to provide quantitative 3D illustrations of induced flow patterns, including vortical structures and vortex core locations. The results reveal that both the magnitude and distribution of induced vortices primarily affect the mixing performance of two viscous flow streams. By using magnetically controlled artificial cilia along with the presented analytical paradigms, a new active flow mixing strategy was suggested to efficiently transport/agitate flows for microfluidics and biomedical applications. PMID:23685964

  5. A novel biosensor to study cAMP dynamics in cilia and flagella

    PubMed Central

    Mukherjee, Shatanik; Jansen, Vera; Jikeli, Jan F; Hamzeh, Hussein; Alvarez, Luis; Dombrowski, Marco; Balbach, Melanie; Strünker, Timo; Seifert, Reinhard; Kaupp, U Benjamin; Wachten, Dagmar

    2016-01-01

    The cellular messenger cAMP regulates multiple cellular functions, including signaling in cilia and flagella. The cAMP dynamics in these subcellular compartments are ill-defined. We introduce a novel FRET-based cAMP biosensor with nanomolar sensitivity that is out of reach for other sensors. To measure cAMP dynamics in the sperm flagellum, we generated transgenic mice and reveal that the hitherto methods determining total cAMP levels do not reflect changes in free cAMP levels. Moreover, cAMP dynamics in the midpiece and principal piece of the flagellum are distinctively different. The sole cAMP source in the flagellum is the soluble adenylate cyclase (SACY). Although bicarbonate-dependent SACY activity requires Ca2+, basal SACY activity is suppressed by Ca2+. Finally, we also applied the sensor to primary cilia. Our new cAMP biosensor features unique characteristics that allow gaining new insights into cAMP signaling and unravel the molecular mechanisms underlying ciliary function in vitro and in vivo. DOI: http://dx.doi.org/10.7554/eLife.14052.001 PMID:27003291

  6. A novel biosensor to study cAMP dynamics in cilia and flagella.

    PubMed

    Mukherjee, Shatanik; Jansen, Vera; Jikeli, Jan F; Hamzeh, Hussein; Alvarez, Luis; Dombrowski, Marco; Balbach, Melanie; Strünker, Timo; Seifert, Reinhard; Kaupp, U Benjamin; Wachten, Dagmar

    2016-01-01

    The cellular messenger cAMP regulates multiple cellular functions, including signaling in cilia and flagella. The cAMP dynamics in these subcellular compartments are ill-defined. We introduce a novel FRET-based cAMP biosensor with nanomolar sensitivity that is out of reach for other sensors. To measure cAMP dynamics in the sperm flagellum, we generated transgenic mice and reveal that the hitherto methods determining total cAMP levels do not reflect changes in free cAMP levels. Moreover, cAMP dynamics in the midpiece and principal piece of the flagellum are distinctively different. The sole cAMP source in the flagellum is the soluble adenylate cyclase (SACY). Although bicarbonate-dependent SACY activity requires Ca(2+), basal SACY activity is suppressed by Ca(2+). Finally, we also applied the sensor to primary cilia. Our new cAMP biosensor features unique characteristics that allow gaining new insights into cAMP signaling and unravel the molecular mechanisms underlying ciliary function in vitro and in vivo. PMID:27003291

  7. Primary ciliary dyskinesia and associated sensory ciliopathies

    PubMed Central

    Horani, Amjad; Ferkol, Thomas W

    2016-01-01

    Primary ciliary dyskinesia (PCD) is a genetic disease of motile cilia, which belongs to a group of disorders resulting from dysfunction of cilia, collectively known as ciliopathies. Insights into the genetics and phenotypes of PCD have grown over the last decade, in part propagated by the discovery of a number of novel cilia-related genes. These genes encode proteins that segregate into structural axonemal, regulatory, as well as cytoplasmic assembly proteins. Our understanding of primary (sensory) cilia has also expanded, and an ever-growing list of diverse conditions has been linked to defective function and signaling of the sensory cilium. Recent multicenter clinical and genetic studies have uncovered the heterogeneity of motile and sensory ciliopathies, and in some cases, the overlap between these conditions. In this review, we will describe the genetics and pathophysiology of ciliopathies in children, focusing on PCD, review emerging genotype-phenotype relationships, and diagnostic tools available for the clinician. PMID:26967669

  8. A mechanism for vertebrate Hedgehog signaling: recruitment to cilia and dissociation of SuFu–Gli protein complexes

    PubMed Central

    Tukachinsky, Hanna; Lopez, Lyle V.

    2010-01-01

    In vertebrates, Hedgehog (Hh) signaling initiated in primary cilia activates the membrane protein Smoothened (Smo) and leads to activation of Gli proteins, the transcriptional effectors of the pathway. In the absence of signaling, Gli proteins are inhibited by the cytoplasmic protein Suppressor of Fused (SuFu). It is unclear how Hh activates Gli and whether it directly regulates SuFu. We find that Hh stimulation quickly recruits endogenous SuFu–Gli complexes to cilia, suggesting a model in which Smo activates Gli by relieving inhibition by SuFu. In support of this model, we find that Hh causes rapid dissociation of the SuFu–Gli complex, thus allowing Gli to enter the nucleus and activate transcription. Activation of protein kinase A (PKA), an inhibitor of Hh signaling, blocks ciliary localization of SuFu–Gli complexes, which in turn prevents their dissociation by signaling. Our results support a simple mechanism in which Hh signals at vertebrate cilia cause dissociation of inactive SuFu–Gli complexes, a process inhibited by PKA. PMID:20956384

  9. Circularly polarised phosphorescent photoluminescence and electroluminescence of iridium complexes

    PubMed Central

    Li, Tian-Yi; Jing, Yi-Ming; Liu, Xuan; Zhao, Yue; Shi, Lin; Tang, Zhiyong; Zheng, You-Xuan; Zuo, Jing-Lin

    2015-01-01

    Nearly all the neutral iridium complexes widely used as dopants in PhOLEDs are racemic mixtures; however, this study observed that these complexes can be separated into stable optically active Λ and ∆ isomers and that their chirality is an intrinsic property. The circularly polarised phosphorescent photoluminescence (CPPPL) signals of Λ/Δ isomers are perfect mirror images with opposite polarisation and equal intensity exhibiting a “handedness” for the polarisation. For the first time, we applied the Λ/Δ iridium isomers as emitters in OLEDs, and the circularly polarised phosphorescent electroluminescence (CPPEL) spectra reveal completely positive or negative broad peaks consistent with the CPPPL spectra. The results demonstrate that the Λ/Δ isomers have potential application for 3D OLEDs because they can exhibit high efficiency and luminance, and 3D display technology based on circularly polarised light is the most comfortable for the eyes. PMID:26446521

  10. The Primary Cilium: A Signaling Center During Vertebrate Development

    PubMed Central

    Goetz, Sarah C.

    2011-01-01

    The primary cilium has recently stepped into the spotlight, as a flood of data demonstrate that this organelle has crucial roles in vertebrate development and human genetic diseases. Cilia are required for the response to developmental signals, and evidence is accumulating that the primary cilium is specialized for Hedgehog (Hh) signal transduction. Formation of cilia, in turn, is regulated by other signaling pathways, possibly including the planar cell polarity pathway. The cilium therefore represents a nexus for signaling pathways during development. The connections between cilia and developmental signaling have begun to clarify the basis of human diseases associated with ciliary dysfunction. PMID:20395968

  11. Cilia-based flow network in the brain ventricles.

    PubMed

    Faubel, Regina; Westendorf, Christian; Bodenschatz, Eberhard; Eichele, Gregor

    2016-07-01

    Cerebrospinal fluid conveys many physiologically important signaling factors through the ventricular cavities of the brain. We investigated the transport of cerebrospinal fluid in the third ventricle of the mouse brain and discovered a highly organized pattern of cilia modules, which collectively give rise to a network of fluid flows that allows for precise transport within this ventricle. We also discovered a cilia-based switch that reliably and periodically alters the flow pattern so as to create a dynamic subdivision that may control substance distribution in the third ventricle. Complex flow patterns were also present in the third ventricles of rats and pigs. Our work suggests that ciliated epithelia can generate and maintain complex, spatiotemporally regulated flow networks. PMID:27387952

  12. Nanoscale Fluidics: Using magnetic nanorods as model cilia

    NASA Astrophysics Data System (ADS)

    Hao, Jing; Ben, Wilde; Jeremy, Cribb; Chris, Dwyer; Jay, Fisher; Kalpit, Desai; Leandra, Vicci; Russell, M. Taylor, II; Richard, Superfine

    2003-11-01

    The beating of cilia and flagella, slender cylinders 250 nanometers in diameter with lengths from 7 to 50 microns, is ubiquitous in biology. The fluid dynamics produced by the cilia or flagella motion is responsible for organism feeding, propulsion, for bacterial clearance in the lungs and for the right-left asymmetry in vertebrates. We are developing a model system for cilia beating through the use of magnetic nanorods. Using anodized aluminum oxide (AAO) membranes as templates, magnetic rods of permalloy with a diameter of 100 and 200 nm have been fabricated. We will describe the details of fabrication and characterization, and discuss methods used to study the hydrodynamic behavior of these nanorods in aqueous solutions under applied magnetic fields. Movies of these nanorods in oscillating 3-D magnetic fields generated by our 3-dimensional force microscopy (3DFM) clearly show bead motion in vortices around the nanorod. Deliberately transporting beads near the rods by laser trap, we can reproducibly study the hydrodynamic behavior around the nanorods and the influence of local rheological properties.

  13. IFT46 plays crucial roles in craniofacial and cilia development.

    PubMed

    Park, Inji; Lee, Hyun-Kyung; Kim, Chowon; Ismail, Tayaba; Kim, Yoo-Kyung; Park, Jeen-Woo; Kwon, Oh-Shin; Kang, Beom Sik; Lee, Dong-Seok; Park, Tae-Joo; Park, Mae-Ja; Choi, Sun-Cheol; Lee, Hyun-Shik

    2016-08-26

    The intraflagellar transport (IFT) system is essential for bidirectional movement of ciliary components from the basal body to the tip beneath the ciliary sheath and is conserved for cilia and flagella formation in most vertebrates. IFT complex A is involved in anterograde trafficking, whereas complex B is involved in retrograde trafficking. IFT46 is well known as a crucial component of IFT complex B, however, its developmental functions are poorly understood. In this study, we investigated the novel functions of IFT46 during vertebrate development, especially, ciliogenesis and neurogenesis, because IFT46 is strongly expressed in both multiciliated cells of epithelial and neural tissues. Knockdown of IFT46 using morpholino microinjections caused shortening of the body axis as well as the formation of fewer and shorter cilia. Furthermore, loss of IFT46 down-regulated the expression of the neural plate and neural tube markers, thus may influence Wnt/planar cell polarity and the sonic hedgehog signaling pathway during neurogenesis. In addition, loss of IFT46 caused craniofacial defects by interfering with cartilage formation. In conclusion, our results depict that IFT46 plays important roles in cilia as well as in neural and craniofacial development. PMID:27320864

  14. Nature-inspired micro-fluidic manipulation using artificial cilia

    NASA Astrophysics Data System (ADS)

    den Toonder, Jaap; de Goede, Judith; Khatavkar, Vinayak; Anderson, Patrick

    2006-11-01

    One particular micro-fluidics manipulation mechanism ``designed'' by nature is that due to a covering of beating cilia over the external surface of micro-organisms (e.g. Paramecium). A cilium can be viewed as a small hair or flexible rod (in protozoa: typical length 10 μm and diameter 0.1 μm) which is attached to the surface. We have developed polymer micro-actuators, made with standard micro-technology processing, which respond to an applied electrical or magnetic field by changing their shape. The shape and size of the polymer actuators mimics that of cilia occurring in nature. We have shown experimentally that, indeed, our artificial cilia can induce significant flow velocities of at least 75 μm/s in a fluid with a viscosity of 10 mPas. In this paper we will give an overview of our activities in developing the polymer actuators and the corresponding technology, show experimental and numerical fluid flow results, and finally assess the feasibility of applying this new and attractive micro-fluidic actuation method in functional biosensors.

  15. Robust estimation of the motile cilia beating frequency.

    PubMed

    Meste, O; Brau, F; Guyon, A

    2015-10-01

    The estimation of the cilia beating frequency (CBF) is of great interest in understanding how the CBF modulates liquid fluxes and how it is controlled by the ciliated cell intra- and/or extracellular medium composition in physiological processes. Motion artifacts and camera defaults may hinder the computation of the frequency variations during long-lasting experiments. We have developed a new analysis approach consisting of a preliminary corrective step (removal of a grid pattern on the image sequence and shift compensation), followed by a harmonic model of the observed cilia using a maximum likelihood estimator framework. It is shown that a more accurate estimation of the frequency can be obtained by averaging the squared Fourier transform of individual pixels followed by a particular summation over the different frequencies, namely the compressed spectrum. Robustness of the proposed method over traditional approaches is shown by several examples and simulations. The method is then applied to images of samples containing ciliated ependymal cells located in the third cerebral ventricle of mouse brains, showing that even small variations in CBF in response to changes in the amount of oxygenation, pH or glucose were clearly visible in the computed frequencies. As a conclusion, this method reveals a fine metabolic tuning of the cilia beating in ependimocytes lining the third cerebral ventricle. Such regulations are likely to participate in homeostatic mechanisms regulating CSF movements and brain energy supply. PMID:26215519

  16. Tubulin glycylases and glutamylases have distinct functions in stabilization and motility of ependymal cilia

    PubMed Central

    Bosch Grau, Montserrat; Gonzalez Curto, Gloria; Rocha, Cecilia; Magiera, Maria M.; Marques Sousa, Patricia; Giordano, Tiziana

    2013-01-01

    Microtubules are subject to a variety of posttranslational modifications that potentially regulate cytoskeletal functions. Two modifications, glutamylation and glycylation, are highly enriched in the axonemes of most eukaryotes, and might therefore play particularly important roles in cilia and flagella. Here we systematically analyze the dynamics of glutamylation and glycylation in developing mouse ependymal cilia and the expression of the corresponding enzymes in the brain. By systematically screening enzymes of the TTLL family for specific functions in ependymal cilia, we demonstrate that the glycylating enzymes TTLL3 and TTLL8 were required for stability and maintenance of ependymal cilia, whereas the polyglutamylase TTLL6 was necessary for coordinated beating behavior. Our work provides evidence for a functional separation of glutamylating and glycylating enzymes in mammalian ependymal cilia. It further advances the elucidation of the functions of tubulin posttranslational modifications in motile cilia of the mammalian brain and their potential importance in brain development and disease. PMID:23897886

  17. On the compatibility of polarisable and non-polarisable models for liquid water

    NASA Astrophysics Data System (ADS)

    Bachmann, Stephan Jan; van Gunsteren, Wilfred F.

    2014-11-01

    The properties of water at physiological conditions can be modelled at different levels of resolution: (1) sub-atomic models that take into account electronic degrees of freedom, (2) atomic models that only account for atomic degrees of freedom and (3) supra-molecular models that only involve some supra-molecular degrees of freedom. To enhance the computational efficiency of molecular simulation, models at different levels of resolution should be simultaneously usable for different parts of a system for which the level of detail of interest is different. This requires these different types of models to be compatible with each other. In the present study, the compatibility of two polarisable models for liquid water, COS/G2 and COS/D, with a non-polarisable model for liquid water, SPC, is investigated. It is shown that these models are compatible. The polarisable models can thus be used to solvate biomolecules described by a biomolecular force field that is compatible with the SPC water model.

  18. The Strange Quark Polarisation from COMPASS data

    SciTech Connect

    Kouznetsov, O.

    2009-12-17

    The strange quark helicity distribution {delta}s(x) was derived at LO from the inclusive asymmetry A{sub a,d} and the semi-inclusive asymmetries A{sub 1,d}{sup {pi}}{sup +}, A{sub 1,d}{sup {pi}}{sup -}, A{sub 1,d}{sup K+}, A{sub 1,d}{sup K-}, measured by COMPASS in polarised deep inelastic muon-deuteron scattering. The distribution of {delta}s(x) is compatible with zero in the whole measured range. The value of the first moment of {delta}s and its error are very sensitive to the assumed value of the ratio of the s-bar-quark to u-quark fragmentation functions into positive kaons {integral}D(K+/s)(z)dz/{integral}D{sub u}{sup K+}(z)dz.

  19. Null point of discrimination in crustacean polarisation vision.

    PubMed

    How, Martin J; Christy, John; Roberts, Nicholas W; Marshall, N Justin

    2014-07-15

    The polarisation of light is used by many species of cephalopods and crustaceans to discriminate objects or to communicate. Most visual systems with this ability, such as that of the fiddler crab, include receptors with photopigments that are oriented horizontally and vertically relative to the outside world. Photoreceptors in such an orthogonal array are maximally sensitive to polarised light with the same fixed e-vector orientation. Using opponent neural connections, this two-channel system may produce a single value of polarisation contrast and, consequently, it may suffer from null points of discrimination. Stomatopod crustaceans use a different system for polarisation vision, comprising at least four types of polarisation-sensitive photoreceptor arranged at 0, 45, 90 and 135 deg relative to each other, in conjunction with extensive rotational eye movements. This anatomical arrangement should not suffer from equivalent null points of discrimination. To test whether these two systems were vulnerable to null points, we presented the fiddler crab Uca heteropleura and the stomatopod Haptosquilla trispinosa with polarised looming stimuli on a modified LCD monitor. The fiddler crab was less sensitive to differences in the degree of polarised light when the e-vector was at -45 deg than when the e-vector was horizontal. In comparison, stomatopods showed no difference in sensitivity between the two stimulus types. The results suggest that fiddler crabs suffer from a null point of sensitivity, while stomatopods do not. PMID:24737768

  20. The strange history of polarised neutrons in Australia

    NASA Astrophysics Data System (ADS)

    Hicks, T. J.

    2016-04-01

    The history of polarised neutrons in Australia is unusual firstly because of the particular access that individuals in universities had to the facilities at the reactor site and because this resulted in the experiments being done almost all being with polarisation analysis. Two instruments were initially available. One was a conventional instrument albeit with a tilting counter. The other was a primitive polarisation analysis instrument purpose built for diffuse scattering. This latter instrument evolved over more than thirty years and produced results ranging from the separation of magnetic and nuclear diffuse scattering, for which it was conceived, to the isolation of magnetic features in inelastic spectra.

  1. The zebrafish fleer gene encodes an essential regulator of cilia tubulin polyglutamylation.

    PubMed

    Pathak, Narendra; Obara, Tomoko; Mangos, Steve; Liu, Yan; Drummond, Iain A

    2007-11-01

    Cilia and basal bodies are essential organelles for a broad spectrum of functions, including the development of left-right asymmetry, kidney function, cerebrospinal fluid transport, generation of photoreceptor outer segments, and hedgehog signaling. Zebrafish fleer (flr) mutants exhibit kidney cysts, randomized left-right asymmetry, hydrocephalus, and rod outer segment defects, suggesting a pleiotropic defect in ciliogenesis. Positional cloning flr identified a tetratricopeptide repeat protein homologous to the Caenorhabditis elegans protein DYF1 that was highly expressed in ciliated cells. flr pronephric cilia were shortened and showed a reduced beat amplitude, and olfactory cilia were absent in mutants. flr cilia exhibited ultrastructural defects in microtubule B-tubules, similar to axonemes that lack tubulin posttranslational modifications (polyglutamylation or polyglycylation). flr cilia showed a dramatic reduction in cilia polyglutamylated tubulin, indicating that flr encodes a novel modulator of tubulin polyglutamylation. We also found that the C. elegans flr homologue, dyf-1, is also required for tubulin polyglutamylation in sensory neuron cilia. Knockdown of zebrafish Ttll6, a tubulin polyglutamylase, specifically eliminated tubulin polyglutamylation and cilia formation in olfactory placodes, similar to flr mutants. These results are the first in vivo evidence that tubulin polyglutamylation is required for vertebrate cilia motility and structure, and, when compromised, results in failed ciliogenesis. PMID:17761526

  2. A facile template-free approach to magnetodriven, multifunctional artificial cilia.

    PubMed

    Timonen, Jaakko V I; Johans, Christoffer; Kontturi, Kyösti; Walther, Andreas; Ikkala, Olli; Ras, Robin H A

    2010-08-01

    Flexible and magnetic artificial cilia were grown on various substrates by a facile bottom-up approach based on template-free magnetic assembly. The magnetic cilia formed spontaneously from a suspension of micrometer-sized ferromagnetic particles and elastomeric polymer in a liquid solvent when dried in an external magnetic field. The cilia mimics were mechanically stable even in the absence of an external magnetic field and a solvent due to the polymer, which acted as "glue" holding the particles together and connecting the cilia to the substrate. The length of the magnetic cilia was in the millimeter range, that is, two to three orders of magnitude times the length of typical biological cilia. The aspect ratio reached values over 100 and was tunable with the magnetic field gradient and the size of the ferromagnetic particles. The cilia mimics responded to an external magnetic field by reversibly bending along the field. The bending actuation was sufficiently powerful to allow two functions: to translate macroscopic nonmagnetic objects placed over the cilia mimics and to mix liquids of even high viscosity. The mechanical properties of the magnetic cilia could be easily tuned by changing the impregnating polymer. The particularly simple template-free construction and fixation on various surfaces suggest applications as an externally controllable surface. PMID:20695442

  3. Exploiting cellophane birefringence to generate radially and azimuthally polarised vector beams

    NASA Astrophysics Data System (ADS)

    Kalwe, Johnston; Neugebauer, Martin; Ominde, Calvine; Leuchs, Gerd; Rurimo, Geoffrey; Banzer, Peter

    2015-03-01

    We exploit the birefringence of cellophane to convert a linearly polarised Gaussian beam into either a radially or azimuthally polarised beam. For that, we fabricated a low-cost polarisation mask consisting of four segments of cellophane. The fast axis of each segment is oriented appropriately in order to rotate the polarisation of the incident linearly polarised beam as desired. To ensure the correct operation of the polarisation mask, we tested the polarisation state of the generated beam by measuring the spatial distribution of the Stokes parameters. Such a device is very cost efficient and allows for the generation of cylindrical vector beams of high quality.

  4. Biochemical studies of olfaction: isolation, characterization, and odorant binding activity of cilia from rainbow trout olfactory rosettes.

    PubMed

    Rhein, L D; Cagan, R H

    1980-08-01

    The role of cilia in recognition of olfactory stimuli has been controversial. Cilia from the intact olfactory rosettes of the rainbow trout Salmo gairdneri were isolated, characterized biochemically, and examined by electron microscopy. The markers studied are those associated with cilia in other organisms. Dynein arms contain Mg2+-AtPase; this enzyme was enriched in the isolated cilia preparation. Guanine nucleotides are associated with the outer microtubule doublets of cilia but adenine nucleotides are not; a substantial enrichment in guanine, relative to adenine, was found in the cilia preparation. Tubulin, the structural protein component of microtubules, occurs in large amounts in cilia. Disc gel electrophoresis indicated tubulin in the cilia preparation. Electron microscopy confirmed the presence of cilia in the isolated preparation. Rainbow trout have an acute sense of smell and many amino acids are odorants to this species. Functional activity of the cilia preparation relevant to odorant recognition was assessed by using binding of radioactively labeled odorant amino acids. L-Alanine, L-serine, L-threonine, L-lysine, and D-alanine bound to the cilia preparation. This study provides direct biochemical evidence that olfactory cilia bind odorant molecules and supports the hypothesis that odorant recognition sites are integral parts of the cilia. PMID:6449006

  5. Polarised stereo endoscope and narrowband detection for minimal access surgery

    PubMed Central

    Clancy, Neil T.; Arya, Shobhit; Qi, Ji; Stoyanov, Danail; Hanna, George B.; Elson, Daniel S.

    2014-01-01

    Polarisation imaging has the potential to provide enhanced contrast based on variations in the optical properties, such as scattering or birefringence, of the tissue of interest. Examining the signal at different wavebands in the visible spectrum also allows interrogation of different depths and structures. A stereo endoscope has been adapted to allow snapshot acquisition of orthogonal linear polarisation images to generate difference of linear polarisation images. These images are acquired in three narrow bands using a triple-bandpass filter and pair of colour cameras. The first in vivo results, acquired during a surgical procedure on a porcine subject, are presented that show wavelength dependent variations in vessel visibility and an increase in contrast under polarised detection. PMID:25574424

  6. Thermodynamics of continuous media with permanent electric polarisation and magnetisation

    NASA Astrophysics Data System (ADS)

    Brechet, Sylvain; Ansermet, Jean-Philippe

    2012-11-01

    The thermodynamics of an electrically charged, multicomponent fluid with permanent electric polarisation, permanent magnetisation and intrinsic vorticity is analysed in the presence of electromagnetic fields with magnetoelectric coupling in the classical limit. Three equations characterising the fluid are derived: a thermostatic equilibrium equation, a reversible and an irreversible thermodynamic evolution equation. These equations are obtained by taking into account the first and second laws of thermodynamics, the chemical reactions, the second law of Newton in translation and in rotation, the local time evolution of the permanent polarisation and the permanent magnetisation, and Maxwell's equations. Explicit expressions for the temperature and the chemical potentials are derived in terms of the electromagnetic fields, the permanent electric polarisation, the permanent magnetisation, the intrinsic vorticity and the magnetoelectric coupling. The analysis of the irreversible thermodynamics yields novel dissipative equations accounting in particular for dielectrophoresis, magnetophoresis, the relaxation of the permanent electric polarisation and the permanent magnetisation, and other properties of electrorheological and magnetorheological fluids.

  7. Integrins are required for cardioblast polarisation in Drosophila

    PubMed Central

    2012-01-01

    Background The formation of a tubular organ, such as the heart, requires the communication of positional and polarity signals between migratory cells. Key to this process is the establishment of a new luminal domain on the cell surface, generally from the apical domain of a migratory cell. This domain will also acquire basal properties, as it will produce a luminal extracellular matrix. Integrin receptors are the primary means of cell adhesion and adhesion signaling with the extracellular matrix. Here we characterise the requirement of Integrins in a genetic model of vasculogenesis, the formation of the heart in Drosophila. Results As with vertebrates, the Drosophila heart arises from lateral mesoderm that migrates medially to meet their contralateral partners, to then assemble a midline vessel. During migration, Integrins are among the first proteins restricted to the presumptive luminal domain of cardioblasts. Integrins are required for normal levels of leading edge membrane motility. Apical accumulation of Integrins is enhanced by Robo, and reciprocally, apicalisation of luminal factors like Slit and Robo requires Integrin function. Integrins may provide a template for the formation of a lumen by stabilising lumen factors like Robo. Subsequent to migration, Integrin is required for normal cardioblast alignment and lumen formation. This phenotype is most readily modified by other mutations that affect adhesion, such as Talin and extracellular matrix ligands. Conclusion Our findings reveal an instructive role for Integrins in communicating polarising information to cells during migration, and during transition to an epithelial tube structure. PMID:22353787

  8. Polarised nuclei for neutron science: recent applications and perspectives

    NASA Astrophysics Data System (ADS)

    Glättli, Hans

    2004-08-01

    Neutron scattering on nuclei is spin dependent, particularly strongly for 1H. The means to achieve large nuclear polarisations and its use for structure analysis or as spin-handling device are reviewed. High resolution (diffraction) as well as low resolution (SANS) measurements can benefit from polarised nuclei by changing selectively the form factors of Bragg reflections or the contrasts (the scattering length density profiles) in SANS. The internal structure of ribosomes and the conformation of polymers in solution have been investigated by this method. A numerical simulation is presented to show the influence of steady-state polarisation of protons on the scattering from a protein-ARN model complex. In addition, a more recent technique, time-resolved SANS is described. It makes use of spatial polarisation gradients created around paramagnetic centres at the onset of nuclear polarisation. Such polarisation domains can enhance considerably the scattering amplitude of free radicals and thus contribute to determine their positions inside a complex protein. Examples of possible future experiments are proposed which combine simultaneously the selectivity of solid-state NMR techniques and neutron scattering.

  9. The sunstone and polarised skylight: ancient Viking navigational tools?

    NASA Astrophysics Data System (ADS)

    Ropars, Guy; Lakshminarayanan, Vasudevan; Le Floch, Albert

    2014-10-01

    Although the polarisation of the light was discovered at the beginning of the nineteenth century, the Vikings could have used the polarised light around the tenth century in their navigation to America, using a 'sunstone' evoked in the Icelandic Sagas. Indeed, the birefringence of the Iceland spar (calcite), a common crystal in Scandinavia, permits a simple observation of the axis of polarisation of the skylight at the zenith. From this, it is possible to guess the azimuth of a hidden Sun below the horizon, for instance. The high sensitivity of the differential method provided by the ordinary and extraordinary beams of calcite at its so-called isotropy point is about two orders higher than that of the best dichroic polariser and permits to reach an accuracy of ±1° for the Sun azimuth (at sunrise and sunset). Unfortunately, due to the relative fragility of calcite, only the so-called Alderney crystal was discovered on board a 16th ancient ship. Curiously, beyond its use as a sunstone by the Vikings, during these last millennia calcite has led to the discovery of the polarisation of the light itself by Malus and is currently being used to detect the atmospheres of exoplanets. Moreover, the differential method for the light polarisation detection is widely used in the animal world.

  10. Optically induced dynamic nuclear spin polarisation in diamond

    NASA Astrophysics Data System (ADS)

    Scheuer, Jochen; Schwartz, Ilai; Chen, Qiong; Schulze-Sünninghausen, David; Carl, Patrick; Höfer, Peter; Retzker, Alexander; Sumiya, Hitoshi; Isoya, Junichi; Luy, Burkhard; Plenio, Martin B.; Naydenov, Boris; Jelezko, Fedor

    2016-01-01

    The sensitivity of magnetic resonance imaging (MRI) depends strongly on nuclear spin polarisation and, motivated by this observation, dynamical nuclear spin polarisation has recently been applied to enhance MRI protocols (Kurhanewicz et al 2011 Neoplasia 13 81). Nuclear spins associated with the 13C carbon isotope (nuclear spin I = 1/2) in diamond possess uniquely long spin lattice relaxation times (Reynhardt and High 2011 Prog. Nucl. Magn. Reson. Spectrosc. 38 37). If they are present in diamond nanocrystals, especially when strongly polarised, they form a promising contrast agent for MRI. Current schemes for achieving nuclear polarisation, however, require cryogenic temperatures. Here we demonstrate an efficient scheme that realises optically induced 13C nuclear spin hyperpolarisation in diamond at room temperature and low ambient magnetic field. Optical pumping of a nitrogen-vacancy centre creates a continuously renewable electron spin polarisation which can be transferred to surrounding 13C nuclear spins. Importantly for future applications we also realise polarisation protocols that are robust against an unknown misalignment between magnetic field and crystal axis.

  11. Polarisation effects in gradient nano-optics

    SciTech Connect

    Erokhin, N S; Shvartsburg, A B; Zueva, Yu M

    2013-09-30

    The spectra of reflection of s- and p-polarised waves from gradient nanocoatings at arbitrary angles of incidence are found within the framework of two exactly solvable models of such coatings. To use the detected spectra in the visible and IR ranges, for different frequencies and coating thicknesses we present the wave reflection coefficients as functions of dimensionless frequencies related to the refractive index gradient of the coating material. It is shown that reflection from the gradient coatings in question is an order of magnitude weaker than reflection from uniform coatings, other parameters of radiation and the reflection system being equal. We report a new exactly solvable model illustrating the specific effect of gradient film optics – the possibility of non-reflective propagation of an s-wave through such a film (an analogue of the Brewster effect). The prospects are shown for the use of gradient nanostructures with different refractive index profiles to fabricate broadband non-reflective coatings. (nanogradient dielectric coatings and metamaterials)

  12. Polarised black holes in AdS

    NASA Astrophysics Data System (ADS)

    Costa, Miguel S.; Greenspan, Lauren; Oliveira, Miguel; Penedones, João; Santos, Jorge E.

    2016-06-01

    We consider solutions in Einstein-Maxwell theory with a negative cosmological constant that asymptote to global AdS 4 with conformal boundary {S}2× {{{R}}}t. At the sphere at infinity we turn on a space-dependent electrostatic potential, which does not destroy the asymptotic AdS behaviour. For simplicity we focus on the case of a dipolar electrostatic potential. We find two new geometries: (i) an AdS soliton that includes the full backreaction of the electric field on the AdS geometry; (ii) a polarised neutral black hole that is deformed by the electric field, accumulating opposite charges in each hemisphere. For both geometries we study boundary data such as the charge density and the stress tensor. For the black hole we also study the horizon charge density and area, and further verify a Smarr formula. Then we consider this system at finite temperature and compute the Gibbs free energy for both AdS soliton and black hole phases. The corresponding phase diagram generalizes the Hawking-Page phase transition. The AdS soliton dominates the low temperature phase and the black hole the high temperature phase, with a critical temperature that decreases as the external electric field increases. Finally, we consider the simple case of a free charged scalar field on {S}2× {{{R}}}t with conformal coupling. For a field in the SU(N ) adjoint representation we compare the phase diagram with the above gravitational system.

  13. A novel broadband circularly polarised monopole antenna

    NASA Astrophysics Data System (ADS)

    Kang, Ding; Cheng, Gao; Tongbin, Yu; Dexin, Qu

    2015-11-01

    A novel broadband circularly polarised (CP) monopole antenna is designed and implemented in this article. The antenna consists of a radiating patch that is composed of an annular-ring linked by a square ring over the corner and a modified ground plane. The broadband property is achieved based on a novel monopole structure that is connected by two perturbed loops, so the CP wave is generated due to the perturbation. Besides, by cutting a rectangular slit and embedding a vertical stub on the ground plane, the impedance and axial-ratio (AR) bandwidths can be greatly enhanced. The measured results reveal that the proposed monopole antenna has an impedance bandwidth of 4.575 GHz from 2.3 to 6.85 GHz, reaching the particularly broad bandwidth of 99.5%. Furthermore, a wide 3-dB AR bandwidth of 34.6% (1.53 GHz, 3.65-5.18 GHz) centred at 4.42 GHz is achieved. The radiation characteristics of the designed antenna are also presented.

  14. Smeared antibranes polarise in AdS

    NASA Astrophysics Data System (ADS)

    Gautason, Fridrik Freyr; Truijen, Brecht; Van Riet, Thomas

    2015-07-01

    In the recent literature it has been questioned whether the local backreaction of antibranes in flux throats can induce a perturbative brane-flux decay. Most evidence for this can be gathered for D6 branes and D p branes smeared over 6 - p compact directions, in line with the absence of finite temperature solutions for these cases. The solutions in the literature have flat worldvolume geometries and non-compact transversal spaces. In this paper we consider what happens when the worldvolume is AdS and the transversal space is compact. We show that in these circumstances brane polarisation smoothens out the flux singularity, which is an indication that brane-flux decay is prevented. This is consistent with the fact that the cosmological constant would be less negative after brane-flux decay. Our results extend recent results on AdS7 solutions from D6 branes to AdS p+1 solutions from D p branes. We show that supersymmetry of the AdS solutions depend on p non-trivially.

  15. Mechanism of olfactory masking in the sensory cilia

    PubMed Central

    Ishida, Hirohiko; Hikichi, Satoshi; Kurahashi, Takashi

    2009-01-01

    Olfactory masking has been used to erase the unpleasant sensation in human cultures for a long period of history. Here, we show a positive correlation between the human masking and the odorant suppression of the transduction current through the cyclic nucleotide–gated (CNG) and Ca2+-activated Cl− (Cl(Ca)) channels. Channels in the olfactory cilia were activated with the cytoplasmic photolysis of caged compounds, and their sensitiveness to odorant suppression was measured with the whole cell patch clamp. When 16 different types of chemicals were applied to cells, cyclic AMP (cAMP)-induced responses (a mixture of CNG and Cl(Ca) currents) were suppressed widely with these substances, but with different sensitivities. Using the same chemicals, in parallel, we measured human olfactory masking with 6-rate scoring tests and saw a correlation coefficient of 0.81 with the channel block. Ringer's solution that was just preexposed to the odorant-containing air affected the cAMP-induced current of the single cell, suggesting that odorant suppression occurs after the evaporation and air/water partition of the odorant chemicals at the olfactory mucus. To investigate the contribution of Cl(Ca), the current was exclusively activated by using the ultraviolet photolysis of caged Ca, DM-nitrophen. With chemical stimuli, it was confirmed that Cl(Ca) channels were less sensitive to the odorant suppression. It is interpreted, however, that in the natural odorant response the Cl(Ca) is affected by the reduction of Ca2+ influx through the CNG channels as a secondary effect. Because the signal transmission between CNG and Cl(Ca) channels includes nonlinear signal-boosting process, CNG channel blockage leads to an amplified reduction in the net current. In addition, we mapped the distribution of the Cl(Ca) channel in living olfactory single cilium using a submicron local [Ca2+]i elevation with the laser photolysis. Cl(Ca) channels are expressed broadly along the cilia. We conclude that

  16. Planck pre-launch status: Expected LFI polarisation capability

    NASA Astrophysics Data System (ADS)

    Leahy, J. P.; Bersanelli, M.; D'Arcangelo, O.; Ganga, K.; Leach, S. M.; Moss, A.; Keihänen, E.; Keskitalo, R.; Kurki-Suonio, H.; Poutanen, T.; Sandri, M.; Scott, D.; Tauber, J.; Valenziano, L.; Villa, F.; Wilkinson, A.; Zonca, A.; Baccigalupi, C.; Borrill, J.; Butler, R. C.; Cuttaia, F.; Davis, R. J.; Frailis, M.; Francheschi, E.; Galeotta, S.; Gregorio, A.; Leonardi, R.; Mandolesi, N.; Maris, M.; Meinhold, P.; Mendes, L.; Mennella, A.; Morgante, G.; Prezeau, G.; Rocha, G.; Stringhetti, L.; Terenzi, L.; Tomasi, M.

    2010-09-01

    We present a system-level description of the Low Frequency Instrument (LFI) considered as a differencing polarimeter, and evaluate its expected performance. The LFI is one of the two instruments on board the ESA Planck mission to study the cosmic microwave background. It consists of a set of 22 radiometers sensitive to linear polarisation, arranged in orthogonally-oriented pairs connected to 11 feed horns operating at 30, 44 and 70 GHz. In our analysis, the generic Jones and Mueller-matrix formulations for polarimetry are adapted to the special case of the LFI. Laboratory measurements of flight components are combined with optical simulations of the telescope to investigate the values and uncertainties in the system parameters affecting polarisation response. Methods of correcting residual systematic errors are also briefly discussed. The LFI has beam-integrated polarisation efficiency >99% for all detectors, with uncertainties below 0.1%. Indirect assessment of polarisation position angles suggests that uncertainties are generally less than 0.5°, and this will be checked in flight using observations of the Crab nebula. Leakage of total intensity into the polarisation signal is generally well below the thermal noise level except for bright Galactic emission, where the dominant effect is likely to be spectral-dependent terms due to bandpass mismatch between the two detectors behind each feed, contributing typically 1-3% leakage of foreground total intensity. Comparable leakage from compact features occurs due to beam mismatch, but this averages to < 5 × 10-4 for large-scale emission. An inevitable feature of the LFI design is that the two components of the linear polarisation are recovered from elliptical beams which differ substantially in orientation. This distorts the recovered polarisation and its angular power spectrum, and several methods are being developed to correct the effect, both in the power spectrum and in the sky maps. The LFI will return a high

  17. Left-right organizer flow dynamics: how much cilia activity reliably yields laterality?

    PubMed

    Sampaio, Pedro; Ferreira, Rita R; Guerrero, Adán; Pintado, Petra; Tavares, Bárbara; Amaro, Joana; Smith, Andrew A; Montenegro-Johnson, Thomas; Smith, David J; Lopes, Susana S

    2014-06-23

    Internal organs are asymmetrically positioned inside the body. Embryonic motile cilia play an essential role in this process by generating a directional fluid flow inside the vertebrate left-right organizer. Detailed characterization of how fluid flow dynamics modulates laterality is lacking. We used zebrafish genetics to experimentally generate a range of flow dynamics. By following the development of each embryo, we show that fluid flow in the left-right organizer is asymmetric and provides a good predictor of organ laterality. This was tested in mosaic organizers composed of motile and immotile cilia generated by dnah7 knockdowns. In parallel, we used simulations of fluid dynamics to analyze our experimental data. These revealed that fluid flow generated by 30 or more cilia predicts 90% situs solitus, similar to experimental observations. We conclude that cilia number, dorsal anterior motile cilia clustering, and left flow are critical to situs solitus via robust asymmetric charon expression. PMID:24930722

  18. c21orf59/kurly Controls Both Cilia Motility and Polarization.

    PubMed

    Jaffe, Kimberly M; Grimes, Daniel T; Schottenfeld-Roames, Jodi; Werner, Michael E; Ku, Tse-Shuen J; Kim, Sun K; Pelliccia, Jose L; Morante, Nicholas F C; Mitchell, Brian J; Burdine, Rebecca D

    2016-03-01

    Cilia are microtubule-based projections that function in the movement of extracellular fluid. This requires cilia to be: (1) motile and driven by dynein complexes and (2) correctly polarized on the surface of cells, which requires planar cell polarity (PCP). Few factors that regulate both processes have been discovered. We reveal that C21orf59/Kurly (Kur), a cytoplasmic protein with some enrichment at the base of cilia, is needed for motility; zebrafish mutants exhibit characteristic developmental abnormalities and dynein arm defects. kur was also required for proper cilia polarization in the zebrafish kidney and the larval skin of Xenopus laevis. CRISPR/Cas9 coupled with homologous recombination to disrupt the endogenous kur locus in Xenopus resulted in the asymmetric localization of the PCP protein Prickle2 being lost in mutant multiciliated cells. Kur also makes interactions with other PCP components, including Disheveled. This supports a model wherein Kur plays a dual role in cilia motility and polarization. PMID:26904945

  19. The BBSome controls IFT assembly and turnaround in cilia.

    PubMed

    Wei, Qing; Zhang, Yuxia; Li, Yujie; Zhang, Qing; Ling, Kun; Hu, Jinghua

    2012-09-01

    The bidirectional movement of intraflagellar transport (IFT) particles, which are composed of motors, IFT-A and IFT-B subcomplexes, and cargoes, is required for the biogenesis and signalling of cilia(1,2). A successful IFT cycle depends on the proper assembly of the massive IFT particle at the ciliary base and its turnaround from anterograde to retrograde transport at the ciliary tip. However, how IFT assembly and turnaround are regulated in vivo remains elusive. From a whole-genome mutagenesis screen in Caenorhabditis elegans, we identified two hypomorphic mutations in dyf-2 and bbs-1 as the only mutants showing normal anterograde IFT transport but defective IFT turnaround at the ciliary tip. Further analyses revealed that the BBSome (refs 3, 4), a group of conserved proteins affected in human Bardet-Biedl syndrome(5) (BBS), assembles IFT complexes at the ciliary base, then binds to the anterograde IFT particle in a DYF-2- (an orthologue of human WDR19) and BBS-1-dependent manner, and lastly reaches the ciliary tip to regulate proper IFT recycling. Our results identify the BBSome as the key player regulating IFT assembly and turnaround in cilia. PMID:22922713

  20. Regulation of cilia assembly, disassembly, and length by protein phosphorylation.

    PubMed

    Cao, Muqing; Li, Guihua; Pan, Junmin

    2009-01-01

    The exact mechanism by which cells are able to assemble, regulate, and disassemble cilia or flagella is not yet completely understood. Recent studies in several model systems, including Chlamydomonas, Tetrahymena, Leishmania, Caenorhabditis elegans, and mammals, provide increasing biochemical and genetic evidence that phosphorylation of multiple protein kinases plays a key role in cilia assembly, disassembly, and length regulation. Members of several protein kinase families--including aurora kinases, never in mitosis A (NIMA)-related protein kinases, mitogen-activated protein (MAP) kinases, and a novel cyclin-dependent protein kinase--are involved in the ciliary regulation process. Among the newly identified protein kinase substrates are Chlamydomonas kinesin-13 (CrKinesin13), a microtubule depolymerizer, and histone deacetylase 6 (HDAC6), a microtubule deacetylase. Chlamydomonas aurora/Ipl1p-like protein kinase (CALK) and CrKinesin13 are two proteins that undergo phosphorylation changes correlated with flagellar assembly or disassembly. CALK becomes phosphorylated when flagella are lost, whereas CrKinesin13 is phosphorylated when new flagella are assembled. Conversely, suppressing CrKinesin13 expression results in cells with shorter flagella. PMID:20362099

  1. Histone deacetylase 6–mediated selective autophagy regulates COPD-associated cilia dysfunction

    PubMed Central

    Lam, Hilaire C.; Cloonan, Suzanne M.; Bhashyam, Abhiram R.; Haspel, Jeffery A.; Singh, Anju; Sathirapongsasuti, J. Fah; Cervo, Morgan; Yao, Hongwei; Chung, Anna L.; Mizumura, Kenji; An, Chang Hyeok; Shan, Bin; Franks, Jonathan M.; Haley, Kathleen J.; Owen, Caroline A.; Tesfaigzi, Yohannes; Washko, George R.; Quackenbush, John; Silverman, Edwin K.; Rahman, Irfan; Kim, Hong Pyo; Mahmood, Ashfaq; Biswal, Shyam S.; Ryter, Stefan W.; Choi, Augustine M.K.

    2013-01-01

    Chronic obstructive pulmonary disease (COPD) involves aberrant airway inflammatory responses to cigarette smoke (CS) that are associated with epithelial cell dysfunction, cilia shortening, and mucociliary clearance disruption. Exposure to CS reduced cilia length and induced autophagy in vivo and in differentiated mouse tracheal epithelial cells (MTECs). Autophagy-impaired (Becn1+/– or Map1lc3B–/–) mice and MTECs resisted CS-induced cilia shortening. Furthermore, CS increased the autophagic turnover of ciliary proteins, indicating that autophagy may regulate cilia homeostasis. We identified cytosolic deacetylase HDAC6 as a critical regulator of autophagy-mediated cilia shortening during CS exposure. Mice bearing an X chromosome deletion of Hdac6 (Hdac6–/Y) and MTECs from these mice had reduced autophagy and were protected from CS-induced cilia shortening. Autophagy-impaired Becn1–/–, Map1lc3B–/–, and Hdac6–/Y mice or mice injected with an HDAC6 inhibitor were protected from CS-induced mucociliary clearance (MCC) disruption. MCC was preserved in mice given the chemical chaperone 4-phenylbutyric acid, but was disrupted in mice lacking the transcription factor NRF2, suggesting that oxidative stress and altered proteostasis contribute to the disruption of MCC. Analysis of human COPD specimens revealed epigenetic deregulation of HDAC6 by hypomethylation and increased protein expression in the airways. We conclude that an autophagy-dependent pathway regulates cilia length during CS exposure and has potential as a therapeutic target for COPD. PMID:24200693

  2. Kinesin-13 regulates the quantity and quality of tubulin inside cilia

    PubMed Central

    Vasudevan, Krishna Kumar; Jiang, Yu-Yang; Lechtreck, Karl F.; Kushida, Yasuharu; Alford, Lea M.; Sale, Winfield S.; Hennessey, Todd; Gaertig, Jacek

    2015-01-01

    Kinesin-13, an end depolymerizer of cytoplasmic and spindle microtubules, also affects the length of cilia. However, in different models, depletion of kinesin-13 either lengthens or shortens cilia, and therefore the exact function of kinesin-13 in cilia remains unclear. We generated null mutations of all kinesin-13 paralogues in the ciliate Tetrahymena. One of the paralogues, Kin13Ap, localizes to the nuclei and is essential for nuclear divisions. The remaining two paralogues, Kin13Bp and Kin13Cp, localize to the cell body and inside assembling cilia. Loss of both Kin13Bp and Kin13Cp resulted in slow cell multiplication and motility, overgrowth of cell body microtubules, shortening of cilia, and synthetic lethality with either paclitaxel or a deletion of MEC-17/ATAT1, the α-tubulin acetyltransferase. The mutant cilia assembled slowly and contained abnormal tubulin, characterized by altered posttranslational modifications and hypersensitivity to paclitaxel. The mutant cilia beat slowly and axonemes showed reduced velocity of microtubule sliding. Thus kinesin-13 positively regulates the axoneme length, influences the properties of ciliary tubulin, and likely indirectly, through its effects on the axonemal microtubules, affects the ciliary dynein-dependent motility. PMID:25501369

  3. Hyperglycemia in the absence of cilia accelerates cystogenesis and induces renal damage.

    PubMed

    Sas, Kelli M; Yin, Hong; Fitzgibbon, Wayne R; Baicu, Catalin F; Zile, Michael R; Steele, Stacy L; Amria, May; Saigusa, Takamitsu; Funk, Jason; Bunni, Marlene A; Siegal, Gene P; Siroky, Brian J; Bissler, John J; Bell, P Darwin

    2015-07-01

    In polycystic kidney disease (PKD), the rate of cyst formation and disease progression is highly variable. The lack of predictability in disease progression may be due to additional environmental factors or pathophysiological processes called "third hits." Diabetes is a growing epidemic, and recent studies suggest that PKD patients may be at an increased risk for this disease. We sought to determine if hyperglycemia enhances the initiation and rate of cystogenesis. Tamoxifen was administered to adult Ift88 conditional floxed allele mice to induce cilia loss in the presence of Cre. Subsequent administration of streptozotocin resulted in equivalent hyperglycemia in cilia(+) and cilia(-) mice. Hyperglycemia with loss of cilia increased the rate of cyst formation and cell proliferation. Structural and functional alterations in the kidney, including focal glomerular foot process effacement, interstitial inflammation, formation of primitive renal tubules, polyuria, and increased proteinuria, were also observed in hyperglycemic cilia(-) mice. Gene array analysis indicated enhanced Wnt and epithelial-to-mesenchymal transition signaling in the kidney of hyperglycemic cilia(-) mice. These data show that hyperglycemia, in the absence of cilia, results in renal structural and functional damage and accelerates cystogenesis, suggesting that diabetes is a risk factor in the progression of PKD. PMID:25904703

  4. Polarisation Transfer in Proton Compton Scattering at High Momentum Transfer

    SciTech Connect

    Hamilton, David

    2004-12-31

    The Jefferson Lab Hall A experiment E99-114 comprised a series of measurements to explore proton Compton scattering at high momentum transfer. For the first time, the polarisation transfer observables in the p (~ 0 ~ p) reaction were measured in the GeV energy range, where it is believed that quark-gluon degrees of freedom begin to dominate. The experiment utilised a circularly polarised photon beam incident on a liquid hydrogen target, with the scattered photon and recoil proton detected in a lead-glass calorimeter and a magnetic spectrometer, respectively.

  5. OPTICAL FIBRES AND FIBREOPTIC SENSORS: Polarisation reflectometry of anisotropic optical fibres

    NASA Astrophysics Data System (ADS)

    Konstantinov, Yurii A.; Kryukov, Igor'I.; Pervadchuk, Vladimir P.; Toroshin, Andrei Yu

    2009-11-01

    Anisotropic, polarisation-maintaining fibres have been studied using a reflectometer and integrated optic polariser. Linearly polarised pulses were launched into the fibre under test at different angles between their plane of polarisation and the main optical axis of the fibre. A special procedure for the correlation analysis of these reflectograms is developed to enhance the reliability of the information about the longitudinal optical uniformity ofanisotropic fibres.

  6. Metachronal wave of artificial cilia array actuated by applied magnetic field

    NASA Astrophysics Data System (ADS)

    Tsumori, Fujio; Marume, Ryuma; Saijou, Akinori; Kudo, Kentaro; Osada, Toshiko; Miura, Hideshi

    2016-06-01

    In this paper, a biomimetic microstructure related to cilia, which are effective fluidic and conveying systems in nature, is described. Authors have already reported that a magnetic elastomer pillar actuated by a rotating magnetic field can work like a natural cilium. In the present work, we show examples of a cilia array with a metachronal wave as the next step. A metachronal wave is a sequential action of a number of cilia. It is theoretically known that a metachronal wave gives a higher fluidic efficiency; however, there has been no report on a metachronal wave by artificial cilia. We prepared magnetic elastomer pillars that contain chainlike clusters of magnetic particles. The orientation of chains was set to be different in each pillar so that each pillar will deform with a different phase.

  7. Structure in pp elastic polarisation and pomeron-flip

    SciTech Connect

    Kamran, M.

    1980-06-01

    An investigation of pp (and np) elastic polarisations in the region 10 less than or equal to p/sub lab/ less than or equal to 300 GeV/c and absolute value of t less than or equal to 0.9 (GeV/c)/sup 2/ is carried out using the Regge pole model with phenomenologically parameterized residues. In an attempt to reproduce the polarisation structure for absolute value of t less than or equal to 0.5 (GeV/c)/sup 2/ and p/sub lab/ greater than or equal to 45 GeV/c a Pomeron-flip contribution is included. It is shown that a Pomeron-flip contribution, although yielding a good overall chi/sup 2//pt for a fit to the data, cannot reproduce the polarisation zero observed in the aforementioned (p/sub lab/,t) region (we use ..cap alpha../sub p/ = 0.23). It is however found that if one invokes an Odderon-flip contribution the polarisation structure can be easily reproduced. The Odderon is the odd-signatured partner of the Pomeron and has isospin one. Its existence has been recently invoked in order to account for the disturbing energy dependence of the np CEX data.

  8. Spin-SILC: CMB polarisation component separation with spin wavelets

    NASA Astrophysics Data System (ADS)

    Rogers, Keir K.; Peiris, Hiranya V.; Leistedt, Boris; McEwen, Jason D.; Pontzen, Andrew

    2016-08-01

    We present Spin-SILC, a new foreground component separation method that accurately extracts the cosmic microwave background (CMB) polarisation E and B modes from raw multifrequency Stokes Q and U measurements of the microwave sky. Spin-SILC is an internal linear combination method that uses spin wavelets to analyse the spin-2 polarisation signal P = Q + iU. The wavelets are additionally directional (non-axisymmetric). This allows different morphologies of signals to be separated and therefore the cleaning algorithm is localised using an additional domain of information. The advantage of spin wavelets over standard scalar wavelets is to simultaneously and self-consistently probe scales and directions in the polarisation signal P = Q + iU and in the underlying E and B modes, therefore providing the ability to perform component separation and E-B decomposition concurrently for the first time. We test Spin-SILC on full-mission Planck simulations and data and show the capacity to correctly recover the underlying cosmological E and B modes. We also demonstrate a strong consistency of our CMB maps with those derived from existing component separation methods. Spin-SILC can be combined with the pseudo- and pure E-B spin wavelet estimators presented in a companion paper to reliably extract the cosmological signal in the presence of complicated sky cuts and noise. Therefore, it will provide a computationally-efficient method to accurately extract the CMB E and B modes for future polarisation experiments.

  9. The leading hadronic vacuum polarisation on the lattice

    SciTech Connect

    Della Morte, Michele; Wittig, Hartmut; Jaeger, Benjamin; Juettner, Andreas

    2011-05-23

    After discussing the relevance of a first principles theory-prediction of the hadronic vacuum polarisation for Standard Model tests, the theoretical challenges for its computation in lattice QCD are reviewed. New ideas that will potentially improve the quality of lattice simulations will be introduced and the status of ongoing simulations will be presented briefly.

  10. Monolithically integrated polarisation mode convertor with a semiconductor laser

    NASA Astrophysics Data System (ADS)

    Naeem, Muhammad Azhar

    In this thesis, the design, optimisation, fabrication and operation of waveguide based semiconductor lasers, integrated with polarisation mode convertors (PMCs), is described. Devices are fabricated in the GaAs/AlGaAs and InP/AlGaInAs material systems, using two types of structures; single PMC and back-to-back PMCs. The convertor designs are based upon air trenches, of sub-wavelength dimensions, being introduced into waveguide structures in order to achieve an asymmetric cross-sectional profile, resulting in wave-pate functionality. The GaAs/AlGaAs PMCs are fabricated using reactive ion etching (RIE), and the phenomena of RIE lag technique is also exploited for obtaining the required asymmetric waveguide profile in a single etch step. These are then integrated with semiconductor lasers. The InP/AlGaInAs PMCs are fabricated using a combination of RIE and inductively coupled plasma (ICP) etching and are integrated with semiconductor lasers and also differential phase shifter (DPS) sections to realise devices with active polarisation control. Integrated devices fabricated on InP/AlGaInAs material system with a semiconductor laser, a PMC followed by a DPS section yields ~40 % polarisation mode conversion whilst the DPS section is held at the transparency condition. Greater than 85 % polarisation mode conversion was also obtained with back to back PMCs, which was complement to the devices fabricated with a single PMC. Furthermore, a first active polarisation controller, monolithically integrated with a semiconductor laser is reported. High speed modulation of the integrated device with 300 Mbps is also demonstrated via current injection to the phase shifter section of the device.

  11. Nucleon polarisabilities at and beyond physical pion masses

    NASA Astrophysics Data System (ADS)

    Grießhammer, Harald W.; McGovern, Judith A.; Phillips, Daniel R.

    2016-05-01

    We examine the results of Chiral Effective Field Theory ( χ EFT) for the scalar- and spin-dipole polarisabilities of the proton and neutron, both for the physical pion mass and as a function of {m_{π}}. This provides chiral extrapolations for lattice QCD polarisability computations. We include both the leading and subleading effects of the nucleon's pion cloud, as well as the leading ones of the Δ(1232)-resonance and its pion cloud. The analytic results are complete at N2LO in the δ counting for pion masses close to the physical value, and at leading order for pion masses similar to the Delta-nucleon mass splitting. In order to quantify the truncation error of our predictions and fits as 68% degree-of-belief intervals, we use a Bayesian procedure recently adapted to EFT expansions. At the physical point, our predictions for the spin polarisabilities are, within respective errors, in good agreement with alternative extractions using experiments and dispersion-relation theory. At larger pion masses we find that the chiral expansion of all polarisabilities becomes intrinsically unreliable as {m_{π}} approaches about 300 MeV -as has already been seen in other observables. χ EFT also predicts a substantial isospin splitting above the physical point for both the electric and magnetic scalar polarisabilities; and we speculate on the impact this has on the stability of nucleons. Our results agree very well with emerging lattice computations in the realm where χ EFT converges. Curiously, for the central values of some of our predictions, this agreement persists to much higher pion masses. We speculate on whether this might be more than a fortuitous coincidence.

  12. Cytoplasmic carboxypeptidase 5 regulates tubulin glutamylation and zebrafish cilia formation and function

    PubMed Central

    Pathak, Narendra; Austin-Tse, Christina A.; Liu, Yan; Vasilyev, Aleksandr; Drummond, Iain A.

    2014-01-01

    Glutamylation is a functionally important tubulin posttranslational modification enriched on stable microtubules of neuronal axons, mitotic spindles, centrioles, and cilia. In vertebrates, balanced activities of tubulin glutamyl ligase and cytoplasmic carboxypeptidase deglutamylase enzymes maintain organelle- and cell type–specific tubulin glutamylation patterns. Tubulin glutamylation in cilia is regulated via restricted subcellular localization or expression of tubulin glutamyl ligases (ttlls) and nonenzymatic proteins, including the zebrafish TPR repeat protein Fleer/Ift70. Here we analyze the expression patterns of ccp deglutamylase genes during zebrafish development and the effects of ccp gene knockdown on cilia formation, morphology, and tubulin glutamylation. The deglutamylases ccp2, ccp5, and ccp6 are expressed in ciliated cells, whereas ccp1 expression is restricted to the nervous system. Only ccp5 knockdown increases cilia tubulin glutamylation, induces ciliopathy phenotypes, including axis curvature, hydrocephalus, and pronephric cysts, and disrupts multicilia motility, suggesting that Ccp5 is the principal tubulin deglutamylase that maintains functional levels of cilia tubulin glutamylation. The ability of ccp5 knockdown to restore cilia tubulin glutamylation in fleer/ift70 mutants and rescue pronephric multicilia formation in both fleer- and ift88-deficient zebrafish indicates that tubulin glutamylation is a key driver of ciliogenesis. PMID:24743595

  13. Spatial organization of cilia tufts governs airways mucus transport: Application to severe asthma

    NASA Astrophysics Data System (ADS)

    Khelloufi, Mustapha Kamel; Gras, Delphine; Chanez, Pascal; Viallat, Annie

    2014-11-01

    We study the coupling between both density and spatial repartition of beating cilia tufts, and the coordinated transport of mucus in an in-vitro epithelial model. We use a fully differentiated model epithelium in air liquid interface (ALI) obtained from endo-bronchial biopsies from healthy subjects and patients with asthma. The asthma phenotype is known to persist in the model. Mucus transport is characterized by the trajectories and velocities of microscopic beads incorporated in the mucus layer. When the beating cilia tufts density is higher than dc = 11/100 × 100 μm2 a spherical spiral coordinated mucus transport is observed over the whole ALI chamber (radius = 6 mm). Below dc, local mucus coordinated transport is observed on small circular domains on the epithelium surface. We reveal that the radii of these domains scale with the beating cilia tufts density with a power 3.7. Surprisingly, this power law is independent on cilia beat frequency, concentration and rheological properties of mucus for healthy subject and patient with asthma. The rotating or linear mucus transport is related to dispersion of the cilia tufts on the epithelium surface. We show that impaired mucus transport observed in severe asthma model epithelia is due to a drastic lack and dysfunction of cilia tufts. The author acknowledges the support of the French Agence Nationale de la Recherche (ANR) under reference ANR-13-BSV5-0015-01.

  14. Directed Fluid Flow Produced by Arrays of Magnetically Actuated Core-Shell Biomimetic Cilia

    NASA Astrophysics Data System (ADS)

    Fiser, B. L.; Shields, A. R.; Evans, B. A.; Superfine, R.

    2010-03-01

    We have developed a novel core-shell microstructure that we use to fabricate arrays of flexible, magnetically actuated biomimetic cilia. Our biomimetic cilia mimic the size and beat shape of biological cilia in order to replicate the transport of fluid driven by cilia in many biological systems including the determination of left-right asymmetry in the vertebrate embryonic nodal plate and mucociliary clearance in the lung. Our core-shell structures consist of a flexible poly(dimethylsiloxane) (PDMS) core surrounded by a shell of nickel approximately forty nanometers thick; by using a core-shell structure, we can tune the mechanical and magnetic properties independently. We present the fabrication process and the long-range transport that occurs above the beating biomimetic cilia tips and will report on progress toward biomimetic cilia induced flow in viscoelastic fluids similar to mucus in the human airway. These flows may have applications in photonics and microfluidics, and our structures may be further useful as sensors or actuators in microelectromechanical systems.

  15. Quantitative description of fluid flows produced by left-right cilia in zebrafish.

    PubMed

    Fox, Craig; Manning, M Lisa; Amack, Jeffrey D

    2015-01-01

    Motile cilia generate directional flows that move mucus through airways, cerebrospinal fluid through brain ventricles, and oocytes through fallopian tubes. In addition, specialized monocilia beat in a rotational pattern to create asymmetric flows that are involved in establishing the left-right (LR) body axis during embryogenesis. These monocilia, which we refer to as "left-right cilia," produce a leftward flow of extraembryonic fluid in a transient "organ of asymmetry" that directs asymmetric signaling and development of LR asymmetries in the cardiovascular system and gastrointestinal tract. The asymmetric flows are thought to establish a chemical gradient and/or activate mechanosensitive cilia to initiate calcium ion signals and a conserved Nodal (TGFβ) pathway on the left side of the embryo, but the mechanisms underlying this process remain unclear. The zebrafish organ of asymmetry, called Kupffer's vesicle, provides a useful model system for investigating LR cilia and cilia-powered fluid flows. Here, we describe methods to visualize flows in Kupffer's vesicle using fluorescent microspheres and introduce a new and freely available MATLAB particle tracking code to quantitatively describe these flows. Analysis of normal and aberrant flows indicates this approach is useful for characterizing flow properties that impact LR asymmetry and may be more broadly applicable for quantifying other cilia flows. PMID:25837391

  16. Single-Polarisation, Single-Frequency, 2 cm Ytterbium-Doped Fibre Laser

    SciTech Connect

    Guan, W.; Marciante, J.R.

    2007-05-23

    A single-polarisation, single-frequency, ytterbium-doped silica fibre laser has been demonstrated in a 2 cm linear cavity. The output power reaches 35 mW with an optical signal-to-noise ratio greater than 65 dB. A polarisation-maintaining fibre Bragg grating is used as the polarisation-dependent reflector to generate the single-polarisation output with the polarisation extinction ratio greater than 20 dB. The laser works stably for two hours under laboratory conditions.

  17. Mutations in DYNC2LI1 disrupt cilia function and cause short rib polydactyly syndrome

    PubMed Central

    Taylor, S. Paige; Dantas, Tiago J.; Duran, Ivan; Wu, Sulin; Lachman, Ralph S.; Nelson, Stanley F.; Cohn, Daniel H.; Vallee, Richard B.; Krakow, Deborah

    2015-01-01

    The short rib polydactyly syndromes (SRPS) are a heterogeneous group of autosomal recessive, perinatal-lethal skeletal disorders characterized primarily by short, horizontal ribs, short limbs, and poly-dactyly. Mutations in several genes affecting intraflagellar transport (IFT) cause SRPS but they do not account for all cases. Here we identify additional SRPS genes and further unravel the functional basis for IFT. We perform whole exome sequencing and identify mutations in a new disease-producing gene, cytoplasmic dynein-2 light intermediate chain 1, DYNC2LI1, segregating with disease in three families. Using primary fibroblasts, we show that DYNC2LI1 is essential for dynein-2 complex stability and that mutations in DYNC2LI1 result in variable-length, including hyperelongated, cilia, Hedgehog pathway impairment, and ciliary IFT accumulations. The findings in this study expand our understanding of SRPS locus heterogeneity and demonstrate the importance of DYNC2LI1 in dynein-2 complex stability, cilium function, Hedgehog regulation, and skeletogenesis. PMID:26077881

  18. The Ciliary Rootlet Maintains Long-Term Stability of Sensory Cilia

    PubMed Central

    Yang, Jun; Gao, Jiangang; Adamian, Michael; Wen, Xiao-Hong; Pawlyk, Basil; Zhang, Luo; Sanderson, Michael J.; Zuo, Jian; Makino, Clint L.; Li, Tiansen

    2005-01-01

    The striated ciliary rootlet is a prominent cytoskeleton originating from basal bodies of ciliated cells. Although a familiar structure in cell biology, its function has remained unresolved. In this study, we carried out targeted disruption in mice of the gene for rootletin, a component of the rootlet. In the mutant, ciliated cells are devoid of rootlets. Phototransduction and ciliary beating in sensory and motile cilia initially exhibit no apparent functional deficits. However, photoreceptors degenerate over time, and mutant lungs appear prone to pathological changes consistent with insufficient mucociliary clearance. Further analyses revealed a striking fragility at the ciliary base in photoreceptors lacking rootlets. In vitro assays suggest that the rootlet is among the least dynamic of all cytoskeletons and interacts with actin filaments. Thus, a primary function of the rootlet is to provide structural support for the cilium. Inasmuch as photoreceptors elaborate an exceptionally enlarged sensory cilium, they are especially dependent on the rootlet for structural integrity and long-term survival. PMID:15870283

  19. Centrin2 regulates CP110 removal in primary cilium formation

    PubMed Central

    Prosser, Suzanna L.

    2015-01-01

    Primary cilia are antenna-like sensory microtubule structures that extend from basal bodies, plasma membrane–docked mother centrioles. Cellular quiescence potentiates ciliogenesis, but the regulation of basal body formation is not fully understood. We used reverse genetics to test the role of the small calcium-binding protein, centrin2, in ciliogenesis. Primary cilia arise in most cell types but have not been described in lymphocytes. We show here that serum starvation of transformed, cultured B and T cells caused primary ciliogenesis. Efficient ciliogenesis in chicken DT40 B lymphocytes required centrin2. We disrupted CETN2 in human retinal pigmented epithelial cells, and despite having intact centrioles, they were unable to make cilia upon serum starvation, showing abnormal localization of distal appendage proteins and failing to remove the ciliation inhibitor CP110. Knockdown of CP110 rescued ciliation in CETN2-deficient cells. Thus, centrin2 regulates primary ciliogenesis through controlling CP110 levels. PMID:25753040

  20. Phosphatase inhibitor 2 promotes acetylation of tubulin in the primary cilium of human retinal epithelial cells

    PubMed Central

    Wang, Weiping; Brautigan, David L

    2008-01-01

    Background Primary cilia are flagella-like projections from the centriole of mammalian cells that have a key role in cell signaling. Human diseases are linked to defects in primary cilia. Microtubules make up the axoneme of cilia and are selectively acetylated and this is thought to contribute to the stability of the structure. However, mechanisms to regulate tubulin acetylation in cilia are poorly understood. Results Endogenous phosphatase inhibitor-2 (I-2) was found concentrated in cilia of human epithelial cells, and was localized to cilia early in the process of formation, prior to the full acetylation of microtubules. Knockdown of I-2 by siRNA significantly reduced the acetylation of microtubules in cilia, without a net decrease in whole cell tubulin acetylation. There was a reduction in the percentage of I-2 knockdown cells with a primary cilium, but no apparent alteration in the cilium length, suggesting no change in microtubule-based transport processes. Inhibition of either histone deacetylases with trichostatin A, or protein phosphatase-1 with calyculin A in I-2 knockdown cells partially rescued the acetylation of microtubules in cilia and the percentage of cells with a primary cilium. Conclusion The regulatory protein I-2 localizes to the primary cilium where it affects both Ser/Thr phosphorylation and is required for full tubulin acetylation. Rescue of tubulin acetylation in I-2 knockdown cells by different chemical inhibitors shows that deacetylases and phosphatases are functionally interconnected to regulate microtubules. As a multifunctional protein, I-2 may link cell cycle progression to structure and stability of the primary cilium. PMID:19036150

  1. The mechanism of self-organized beating of cilia

    NASA Astrophysics Data System (ADS)

    Vidyadharan, Jyothish Sulochana

    The internal structure and physical properties of cilia are well known. The relevant hydrodynamics is also well known. But the mechanism behind the coordinated activity of the dynein molecular motors is not known. Based on experimental observations, it has been concluded that this mechanism cannot be due to control from the cell body. The possible mechanism has to be self-organized and the trigger for motor activation/deactivation has to be something related to the geometry of the ciliary axoneme. This thesis critically evaluates the most widely currently cited models and suggests an alternative model for how cilia beat. From the literature we obtained wave forms of ciliary beating at different instants in the beat cycle. These instants were digitized and interpolated. From this data, we were able to calculate the hydrodynamic force distribution (external force distribution) on the cilia and the translational and rotational velocities of the cell body. Once the hydrodynamic force distribution was obtained, we calculated the internal force distribution in the cilium using an equation we derived. Once this was known, we were able to calculate parameters of the ciliary axoneme such as the dynamic stiffness. The stiffness is the ratio of the first Fourier modes of the internal force distribution and the relative sliding between the doublet microtubules that form the axoneme. We found that the first mode was the dominant one and is the one we used for calculations. We were also able to calculate the energy involved in formation and propagation of the wave that produces the ciliary beating. We discovered that the dynamic stiffness varies along the length of a cilium. We determined that in the central region of the cilium, the stiffness is almost purely imaginary which means that the sliding velocity follows the internal force generation in that region rather than sliding. We also found that in Fourier space, the flexural rigidity (kappa=EI where E is Young's modulus and

  2. Primary ciliary dyskinesia: cytological and clinical features.

    PubMed

    Greenstone, M; Rutman, A; Dewar, A; Mackay, I; Cole, P J

    1988-05-01

    Thirty patients with functional and/or morphological abnormalities of respiratory tract cilia were identified. The diagnosis of primary ciliary dyskinesia was based on observed abnormalities of ciliary ultrastructure or beating in vitro (beat pattern, beat frequency or percentage of motile cilia). Beat frequency and motility indices approached the normal range in some cases and suggests that the term 'immotile cilia syndrome' is not appropriate. Morphological abnormalities were most commonly due to deficiency of dynein arms, affecting the outer arms (n = 7), inner arms (n = 3) or both (n = 10). Examples of radial spoke and microtubular defects were also identified but in seven subjects ciliary ultrastructure was normal. In six patients paired samples of nasal and bronchial cilia were obtained and showed consistent abnormalities of motility and ultrastructure. Adenosine triphosphate and adenosine triphosphatase did not restore in vitro motility when added to dynein deficient cilia. The clinical picture was of life-long sinusitis and recurrent bronchial infection but the spectrum was broader than that encompassed by Kartagener's triad (dextrocardia, sinusitis and bronchiectasis). Fourteen patients had normal cardiac situs and definite or highly suggestive evidence of bronchiectasis was present in only 17 patients. Radiological evidence of sinusitis was common but absence of frontal sinuses was not universal. Chronic serous otitis media was a frequent finding but deafness was rarely profound. Fertility problems were common but were not universal in female subjects. Lung function testing revealed evidence of airflow obstruction but this was mild in most cases. PMID:2975807

  3. Polarisation properties of Milky-Way-like galaxies

    NASA Astrophysics Data System (ADS)

    Sun, X. H.; Reich, W.

    2012-07-01

    Aims: We study the polarisation properties, magnetic field strength, and synchrotron emission scale-height of Milky-Way-like galaxies in comparison with other spiral galaxies. Methods: We used our 3D-emission model of the Milky Way Galaxy for viewing the Milky Way from outside at various inclinations in the way that spiral galaxies are observed. We analysed these Milky Way maps with techniques used to obtain the strength of magnetic fields, rotation measures (RMs), and scale-heights of synchrotron emission from observations of resolved galaxies and compared the results with the Milky Way model parameter. We also simulated a large sample of unresolved Milky-Way-like galaxies to study their statistical polarisation properties. Results: When seen edge-on, the synchrotron emission from the Milky Way has an exponential scale-height of about 0.74 kpc, which is much lower than the values obtained from previous models. We find that current analysis methods overestimate the scale-height of synchrotron emission of galaxies by about 10% at an inclination of 80° and about 40% at an inclination of 70° because of contamination from the disk. The observed RMs for face-on galaxies derived from high-frequency polarisation measurements approximate to the Faraday depths (FDs) when scaled by a factor of two. For edge-on galaxies, the observed RMs are indicative of the orientation of the large-scale magnetic field, but are not closely related with the FDs. Assuming energy equipartition between the magnetic field and particles for the Milky Way results in an average magnetic-field strength that is about twice as high as the intrinsic value for a K factor of 100. The number distribution of the integrated polarisation percentages of a large sample of unresolved Milky-Way-like galaxies peaks at about 4.2% at 4.8 GHz and at about 0.8% at 1.4 GHz. Integrated polarisation angles rotated by 90° align very well with the position angles of the major axes, implying that unresolved galaxies do

  4. Ectopic cilia associated with an orbital dermoid cyst and sinus tract: case report.

    PubMed

    Krahulík, David; Karhanová, Marta; Vaverka, Miroslav; Brychtová, Světlana; Pospíšilová, Dagmar

    2015-08-01

    Ectopic cilia are extremely rare congenital anomalies in which eyelash follicles appear in an abnormal place on the eyelid, most typically on the lateral quadrant of the anterior surface of the upper eyelid. In the majority of cases, simple surgical excision of ectopic cilia is indicated because of its cosmetic aspect. There is usually no associated medical co-morbidity with this anomaly. The authors report an unusual case of ectopic cilia associated with an orbital dermoid cyst and sinus tract. A 3-year-old boy was initially diagnosed with ectopic cilia on the left upper eyelid. There was no history of inflammation or swelling of the eyelid. An ophthalmological examination revealed only 1 mm of ptosis; no proptosis, inferior displacement, or palpable orbital mass was present. During surgical excision of the ectopic cilia, a thin sinus tract was identified, leading posteriorly to the orbit. Magnetic resonance imaging performed after the excision showed a supraorbital extraconal mass just below the roof of the left orbit. A supraorbital 2-piece craniotomy was performed with total extirpation of the dermoid cyst. The cyst was removed en bloc without damage to the extraocular muscles, but the sinus tract could no longer be identified. Follow-up MRI was performed 6 months after surgery and showed no evidence of recurrence. A follow-up ophthalmological examination showed no signs of inferior displacement or proptosis. To the best of the authors' knowledge, this case is the first reported instance of ectopic cilia associated with a dermoid cyst and sinus tract in which no typical clinical signs and symptoms of possible orbital pathology were present. This case highlights the value of radiological examination in all cases of ectopic cilia prior to surgical excision. PMID:25978533

  5. Electron Impact Excitation of Polarised Sodium and Potassium.

    NASA Astrophysics Data System (ADS)

    Bukhari, Muhammad Abdul-Hakim

    Available from UMI in association with The British Library. Requires signed TDF. A beam of unpolarised electrons is crossed at right angles with a beam of partially polarised sodium or potassium atoms (degree of polarisation { bf P}_{bf A} = 21%). The resonance radiation from excited ^2P_{3/2,1/2} states is observed in a direction perpendicular to both beams, and the Stokes parameters {bf P}_ {bf 1}, {bf P }_{bf 2} and {bf P}_{bf 3} of the emitted photons are measured, {bf P}_{bf 1} = {I(0^circ)-I(90^circ)over I(0^circ)+I(90^circ)}, {bf P}_{bf 2} = {I(45^circ)-I(135^circ)over I(45^circ)+I(135^circ)}, {bf P}_{bf 3} = {I(RHC)-I(LHC)over I(RHC)+I(LHC)} where 0^circ, 45 ^circ, 90^circ and 135^circ corresponds to the polariser angle measured from the electron beam direction and RHC, LHC to right and left hand circular polarisation in the spectroscopic definition. Because of reflection symmetries, only {bf P }_{bf 1} is expected to be non-zero for unpolarised atoms. If polarised atoms are used with their spins parallel to the direction of the observed photons, {bf P}_ {bf 3} should also become non-zero (in addition to {bf P}_ {bf 1}) while {bf P}_{bf 2} remains zero. We report here an independent measurement on the neutral sodium NaI (3^2{bf P }_{3/2,1/2} - 3^2{bf S}_{1/2}) transition ( lambda=589.0, 589.6 nm) and present new results for the neutral potassium KI (5^2{ bf P}_{3/2,1/2} - 4^2 {bf S}_{1/2}) transition (lambda=404.4, 404.7 nm). The potassium results for {bf P}_{ bf 2} agree well with the theoretical prediction of zero and the results for {bf P}_{bf 1} show close similarities to those of sodium results at high and intermediate energies, while at low energy especially close to threshold they differ. The threshold polarisation ({ bf P}_{bf 1}) in the case of potassium is about 10%. Our { bf P}_{bf 3} values for sodium are somewhat higher than the results of Osimitsch and Jitschin. The trend of {bf P} _{bf 3} for potassium is similar to that of sodium in such a

  6. Measurement of the tau polarisation at the Z resonance

    NASA Astrophysics Data System (ADS)

    Buskulic, D.; Decamp, D.; Goy, C.; Lees, J.-P.; Minard, M.-N.; Mours, B.; Pietrzyk, B.; Alemany, R.; Ariztizabal, F.; Comas, P.; Crespo, J. M.; Delfino, M.; Fenandez, E.; Fernandez-Bosman, M.; Gaitan, V.; Garrido, Ll.; Mattison, T.; Pacheco, A.; Padilla, C.; Pascual, A.; Creanza, D.; de Palma, M.; Farilla, A.; Iaselli, G.; Maggi, G.; Maggi, M.; Natali, S.; Nuzzo, S.; Quattromini, M.; Ranieri, A.; Raso, G.; Romano, F.; Ruggieri, F.; Selvaggi, G.; Silvestris, L.; Tempesta, P.; Zito, G.; Chai, Y.; Hu, H.; Huang, D.; Huang, X.; Lin, J.; Wang, T.; Xie, Y.; Xu, D.; Xu, R.; Zhang, J.; Zhao, W.; Bauerdick, L. A. T.; Blucher, E.; Bonvicini, G.; Boudreau, J.; Casper, D.; Drevermann, H.; Forty, R. W.; Ganis, G.; Gay, C.; Hagelberg, R.; Harvey, J.; Haywood, S.; Hilgart, J.; Jacobsen, R.; Jost, B.; Knobloch, J.; Lehraus, I.; Lohse, T.; Lusiani, A.; Martinez, M.; Mato, P.; Meinhard, H.; Minten, A.; Miotto, A.; Miquel, R.; Moser, H.-G.; Palazzi, P.; Perlas, J. A.; Pusztaszeri, J.-F.; Ranjard, F.; Redlinger, G.; Rolandi, L.; Rothberg, J.; Ruan, T.; Saich, M.; Schlatter, D.; Schmelling, M.; Sefkow, F.; Tejessy, W.; Wachsmuth, H.; Wiedenmann, W.; Wildish, T.; Witzeling, W.; Wotschack, J.; Ajaltouni, Z.; Badaud, F.; Bardadin-Otwinowska, M.; El Fellous, R.; Falvard, A.; Gay, P.; Guicheney, C.; Henrard, P.; Jousset, J.; Michel, B.; Montret, J.-C.; Pallin, D.; Perret, P.; Podlyski, F.; Proriol, J.; Prulhière, F.; Saadi, F.; Fearnley, T.; Hansen, J. D.; Hansen, J. R.; Hansen, P. H.; Møllerud, R.; Nilsson, B. S.; Candlin, D. J.; Parsons, M. I.; Veitch, E.; Moneta, L.; Parrini, G.; Corden, M.; Georgiopoulos, C.; Ikeda, M.; Lannutti, J.; Levinthal, D.; Mermikides, M.; Sawyer, L.; Wasserbaech, S.; Antonelli, A.; Baldini, R.; Bencivenni, G.; Bologna, G.; Bossi, F.; Campana, P.; Capon, G.; Cerutti, F.; Chiarella, V.; D'Ettorre-Piazzoli, B.; Felici, G.; Laurelli, P.; Mannocchi, G.; Murtas, F.; Murtas, G. P.; Passalacqua, L.; Pepe-Altarelli, M.; Picchi, P.; Colrain, P.; Ten Have, I.; Lynch, J. G.; Maitland, W.; Morton, W. T.; Raine, C.; Reeves, P.; Scarr, J. M.; Smith, K.; Smith, M. G.; Thompson, A. S.; Turnbull, R. M.; Brandl, B.; Braun, O.; Geweniger, C.; Hanke, P.; Hepp, V.; Kluge, E. E.; Maumary, Y.; Putzer, A.; Rensch, B.; Stahl, A.; Tittel, K.; Wunsch, M.; Belk, A. T.; Beuselinck, R.; Binnie, D. M.; Cameron, W.; Cattaneo, M.; Colling, D. J.; Dornan, P. J.; Dugeay, S.; Greene, A. M.; Hassaed, J. F.; Lieske, N. M.; Nash, J.; Payne, D. G.; Phillips, M. J.; Sedgbeer, J. K.; Tomalin, I. R.; Wright, A. G.; Girtler, P.; Kneringer, E.; Kuhn, D.; Rudolph, G.; Bowdery, C. K.; Brodbeck, T. J.; Finch, A. J.; Foster, F.; Hughes, G.; Jackson, D.; Keemer, N. R.; Nuttall, M.; Patel, A.; Sloan, T.; Snow, S. W.; Whelan, E. P.; Efthymiopoulos, I.; Kyriakis, A.; Simopoulou, E.; Vayaki, A.; Zachariadou, K.; Badier, J.; Blondel, A.; Bonneaud, G.; Brient, J. C.; Fouque, G.; Orteu, S.; Rougé, A.; Rumpf, M.; Tanaka, R.; Verderi, M.; Videau, H.; Adlung, S.; Assmann, R.; Bauer, C.; Blum, W.; Brown, D.; Cattaneo, P.; Dehning, B.; Dietl, H.; Dydak, F.; Frank, M.; Halley, A. W.; Lauber, J.; Lütjens, G.; Lutz, G.; Männer, W.; Richter, R.; Rotscheidt, H.; Schröder, J.; Schwarz, A. S.; Settles, R.; Seywerd, H.; Stierlin, U.; Stiegler, U.; Denis, R. St.; Wolf, G.; Boucrot, J.; Callot, O.; Cordier, A.; Davier, M.; Duflot, L.; Grivaz, J.-F.; Heusse, Ph.; Jaffe, D. E.; Janot, P.; Kim, D. W.; Le Diberder, F.; Lefrançois, J.; Lutz, A.-M.; Schune, M.-H.; Veillet, J.-J.; Videau, I.; Zhang, Z.; Abbaneo, D.; Bagliesi, G.; Batignani, G.; Bosisio, L.; Bottigli, U.; Bozzi, C.; Calderini, G.; Carpinelli, M.; Ciocci, M. A.; Dell'Orso, R.; Ferrante, I.; Fidecaro, F.; Foa, L.; Focardi, E.; Forti, F.; Giassi, A.; Giorgi, M. A.; Gregorio, A.; Ligabue, F.; Mannelli, E. B.; Marrocchesi, P. S.; Messineo, A.; Palla, F.; Rizzo, G.; Sanguinetti, G.; Spagnolo, P.; Steinberger, J.; Tenchini, R.; Tonelli, G.; Triggiani, G.; Vannini, C.; Venturi, A.; Verdini, P. G.; Walsh, J.; Betteridge, A. P.; Carter, J. M.; Green, M. G.; March, P. V.; Mir, Ll. M.; Medcalf, T.; Quazi, I. S.; Strong, J. A.; West, L. R.; Botterill, D. R.; Clifft, R. W.; Edgecock, T. R.; Edwards, M.; Fisher, S. M.; Jones, T. J.; Norton, P. R.; Salmon, D. P.; Thompson, J. C.; Kleinknecht, K.; Raab, J.; Renk, B.; Sander, H.-G.; Schmidt, H.; Steeg, F.; Walther, S. M.; Wanke, R.; Wolf, B.; Aubert, J.-J.; Bencheikh, A. M.; Benchouk, C.; Bonissent, A.; Carr, J.; Coyle, P.; Drinkard, J.; Etienne, F.; Nicod, D.; Papalexiou, S.; Payre, P.; Roos, L.; Rousseau, D.; Schwemling, P.; Talby, M.; Bloch-Devaux, B.; Colas, P.; Duarte, H.; Kozanecki, W.; Lançon, E.; Lemaire, M. C.; Locci, E.; Perez, P.; Perrier, F.; Rander, J.; Renardy, J.-F.; Rosowsky, A.; Roussarie, A.; Schuller, J.-P.; Schwindling, J.; Si Mohand, D.; Vallage, B.; Johnson, R. P.; Litke, A. M.; Taylor, G.; Wear, J.; Ashman, J. G.; Babbage, W.; Booth, C. N.; Buttar, C.; Carney, R. E.; Cartwright, S.; Combley, F.; Hatfield, F.; Thompson, L. F.; Barberio, E.; Böhrer, A.; Brandt, S.; Cowan, G.; Grupen, C.; Lutters, G.; Rivera, F.; Schäfer, U.; Della Marina, R.; Giannini, G.; Gobbo, B.; Ragusa, F.; Bellantoni, L.; Chen, W.; Cinabro, D.; Conway, J. S.; Cowen, D. F.; Feng, Z.; Ferguson, D. P. S.; Gao, Y. S.; Grahl, J.; Harton, J. L.; Jared, R. C.; Leclaire, B. W.; Lishka, C.; Pan, Y. B.; Pater, J. R.; Saadi, Y.; Schmitt, M.; Sharma, V.; Shi, Z. H.; Walsh, A. M.; Weber, F. V.; Wu, Sau Lan; Wu, X.; Zheng, M.; Zobernig, G.

    1993-09-01

    Using 18.8 pb-1 of data collected in 1990 and 1991, ALEPH has measured the tau polarisation in the decay modes τ→ ev bar v, τ→μ v bar v, τ→πν, τ→ρν and τ→ a 1ν, using both the individual tau decay kinematics and the event acollinearity. The measurement of the tau polarisation as a function of the production polar angle yields the two parameters A τ and A e , where A l =2 g {/v l } g {/A l }/( g {/v l })2+( g {/A l })2] The results A τ=0.143±0.023 and A e =0.120±0.026 are consistent with the hypothesis of electron-tau universality. Assuming universality yields a measurement of the effective weak mixing angle sin2θ{/w eff}=0.2332±0.0022.

  7. Pulsar polarisation below 200 MHz: Average profiles and propagation effects

    NASA Astrophysics Data System (ADS)

    Noutsos, A.; Sobey, C.; Kondratiev, V. I.; Weltevrede, P.; Verbiest, J. P. W.; Karastergiou, A.; Kramer, M.; Kuniyoshi, M.; Alexov, A.; Breton, R. P.; Bilous, A. V.; Cooper, S.; Falcke, H.; Grießmeier, J.-M.; Hassall, T. E.; Hessels, J. W. T.; Keane, E. F.; Osłowski, S.; Pilia, M.; Serylak, M.; Stappers, B. W.; ter Veen, S.; van Leeuwen, J.; Zagkouris, K.; Anderson, K.; Bähren, L.; Bell, M.; Broderick, J.; Carbone, D.; Cendes, Y.; Coenen, T.; Corbel, S.; Eislöffel, J.; Fender, R.; Garsden, H.; Jonker, P.; Law, C.; Markoff, S.; Masters, J.; Miller-Jones, J.; Molenaar, G.; Osten, R.; Pietka, M.; Rol, E.; Rowlinson, A.; Scheers, B.; Spreeuw, H.; Staley, T.; Stewart, A.; Swinbank, J.; Wijers, R.; Wijnands, R.; Wise, M.; Zarka, P.; van der Horst, A.

    2015-04-01

    Aims: We present the highest-quality polarisation profiles to date of 16 non-recycled pulsars and four millisecond pulsars, observed below 200 MHz with the LOFAR high-band antennas. Based on the observed profiles, we perform an initial investigation of expected observational effects resulting from the propagation of polarised emission in the pulsar magnetosphere and the interstellar medium. Methods: The polarisation data presented in this paper have been calibrated for the geometric-projection and beam-shape effects that distort the polarised information as detected with the LOFAR antennas. We have used RM Synthesis to determine the amount of Faraday rotation in the data at the time of the observations. The ionospheric contribution to the measured Faraday rotation was estimated using a model of the ionosphere. To study the propagation effects, we have compared our low-frequency polarisation observations with archival data at 240, 400, 600, and 1400 MHz. Results: The predictions of magnetospheric birefringence in pulsars have been tested using spectra of the pulse width and fractional polarisation from multifrequency data. The derived spectra offer only partial support for the expected effects of birefringence on the polarisation properties, with only about half of our sample being consistent with the model's predictions. It is noted that for some pulsars these measurements are contaminated by the effects of interstellar scattering. For a number of pulsars in our sample, we have observed significant variations in the amount of Faraday rotation as a function of pulse phase, which is possibly an artefact of scattering. These variations are typically two orders of magnitude smaller than that observed at 1400 MHz by Noutsos et al. (2009), for a different sample of southern pulsars. In this paper we present a possible explanation for the difference in magnitude of this effect between the two frequencies, based on scattering. Finally, we have estimated the magnetospheric

  8. Measurement of double polarisation asymmetries in ω-photoproduction

    NASA Astrophysics Data System (ADS)

    Eberhardt, H.; Jude, T. C.; Schmieden, H.; Anisovich, A. V.; Bantes, B.; Bayadilov, D.; Beck, R.; Beloglazov, Yu.; Bichow, M.; Böse, S.; Brinkmann, K.-Th.; Challand, Th.; Crede, V.; Diez, F.; Drexler, P.; Dutz, H.; Elsner, D.; Ewald, R.; Fornet-Ponse, K.; Friedrich, St.; Frommberger, F.; Funke, Ch.; Gottschall, M.; Gridnev, A.; Grüner, M.; Gutz, E.; Hammann, Ch.; Hannappel, J.; Hartmann, J.; Hillert, W.; Hoffmeister, Ph.; Honisch, Ch.; Jaegle, I.; Kaiser, D.; Kalinowsky, H.; Kalischewski, F.; Kammer, S.; Keshelashvili, I.; Kleber, V.; Klein, F.; Klempt, E.; Koop, K.; Krusche, B.; Kube, M.; Lang, M.; Lopatin, I.; Maghrbi, Y.; Makonyi, K.; Metag, V.; Meyer, W.; Müller, J.; Nanova, M.; Nikonov, V.; Novotny, R.; Piontek, D.; Reeve, S.; Reicherz, G.; Rostomyan, T.; Runkel, S.; Sarantsev, A.; Schaepe, St.; Schmidt, Ch.; Schmitz, R.; Seifen, T.; Sokhoyan, V.; Sumachev, V.; Thiel, A.; Thoma, U.; Urban, M.; van Pee, H.; Walther, D.; Wendel, Ch.; Wiedner, U.; Wilson, A.; Winnebeck, A.

    2015-11-01

    The first measurements of the beam-target-helicity-asymmetries E and G in the photoproduction of ω-mesons off protons at the CBELSA/TAPS experiment are reported. E (G) was measured using circularly (linearly) polarised photons and a longitudinally polarised target. E was measured over the photon energy range from close to threshold (Eγ = 1108 MeV) to Eγ = 2300 MeV and G at a single energy interval of 1108

  9. Threshold π0 photoproduction on transverse polarised protons at MAMI

    NASA Astrophysics Data System (ADS)

    Schumann, S.; Otte, P. B.; Akondi, C. S.; Annand, J. R. M.; Arends, H.-J.; Beck, R.; Bernstein, A. M.; Borisov, N.; Braghieri, A.; Briscoe, W. J.; Cherepnya, S.; Collicott, C.; Costanza, S.; Downie, E. J.; Dieterle, M.; Fernández-Ramírez, C.; Fix, A.; Fil'kov, L. V.; Garni, S.; Glazier, D. I.; Gradl, W.; Gurevich, G.; Hall-Barrientos, P.; Hamilton, D.; Hornidge, D.; Howdle, D.; Huber, G. M.; Kashevarov, V. L.; Keshelashvili, I.; Kondratiev, R.; Korolija, M.; Krusche, B.; Lazarev, A.; Lisin, V.; Livingston, K.; MacGregor, I. J. D.; Mancel, J.; Manley, D. M.; Martel, P. P.; McNicoll, E. F.; Meyer, W.; Middleton, D.; Miskimen, R.; Mushkarenkov, A.; Nefkens, B. M. K.; Neganov, A.; Nikolaev, A.; Oberle, M.; Ostrick, M.; Ortega, H.; Ott, P.; Oussena, B.; Paudyal, D.; Pedroni, P.; Polonski, A.; Polyanski, V. V.; Prakhov, S.; Reicherz, G.; Rostomyan, T.; Sarty, A.; Sikora, M. H.; Sokhoyan, V.; Steffen, O.; Strakovsky, I. I.; Strub, Th.; Supek, I.; Tiator, L.; Thomas, A.; Unverzagt, M.; Usov, Yu. A.; Watts, D. P.; Werthmüller, D.; Witthauer, L.; Wolfes, M.

    2015-11-01

    Polarisation-dependent differential cross sections σT associated with the target asymmetry T have been measured for the reaction γ p → → pπ0 with transverse target polarisation from π0 threshold to photon energies of 190 MeV. The data were obtained using a frozen-spin butanol target with the Crystal Ball / TAPS detector set-up and the Glasgow photon tagging system at the Mainz Microtron MAMI. Results for σT have been used in combination with our previous measurements of the unpolarised cross section σ0 and the beam asymmetry Σ for a model-independent determination of S- and P-wave multipoles in the π0 threshold region, which includes for the first time a direct determination of the imaginary part of the E0+ multipole.

  10. Prospects for studying vacuum polarisation using dipole and synchrotron radiation

    NASA Astrophysics Data System (ADS)

    Ilderton, Anton; Marklund, Mattias

    2016-02-01

    The measurement of vacuum polarisation effects, in particular vacuum birefringence, using combined optical and X-ray laser pulses are now actively pursued. Here we briefly examine the feasibility of two alternative set-ups. The first utilises an alternative target, namely a converging dipole pulse, and the second uses an alternative probe, namely the synchrotron-like emission from highly energetic particles, themselves interacting with a laser pulse. The latter set-up has been proposed for experiments at ELI-NP.

  11. Review of Recent Calculations of the Hadronic Vacuum Polarisation Contribution

    NASA Astrophysics Data System (ADS)

    Zhang, Zhiqing

    2016-04-01

    Recent calculations of the hadronic vacuum polarisation contribution are reviewed. The focus is put on the leading-order contribution to the muon magnetic anomaly involving e+e- annihilation cross section data as input to a dispersion relation approach. Alternative calculation including tau data is also discussed. The τ data are corrected for various isospin-breaking sources which are explicitly shown source by source.

  12. Polarisation mode dispersion correlations with the coarse-step method

    NASA Astrophysics Data System (ADS)

    Braimiotis, Christos; Eberhard, Marc; Blow, Keith

    2006-06-01

    Having a fixed differential-group delay (DGD) term b‧ in the coarse-step method results in a repetitive pattern in the autocorrelation function (ACF). We solve this problem by inserting a varying DGD term at each integration step. Furthermore we compute the range of values needed for b‧ and simulate the phenomenon of polarisation mode dispersion for different statistical distributions of b‧. We examine systematically the modified coarse-step method compared to the analytical model, through our simulation results.

  13. Ultrafast polarisation spectroscopy of photoinduced charges in a conjugated polymer

    SciTech Connect

    Bakulin, A A; Loosdrecht, P van; Pshenichnikov, M S; Parashchuk, D Yu

    2009-07-31

    Tunable optical parametric generators and amplifiers (OPA), proposed and developed by Akhmanov and his colleagues, have become the working horses in exploration of dynamical processes in physics, chemistry, and biology. In this paper, we demonstrate the possibility of using ultrafast polarisation-sensitive two-colour spectroscopy, performed with a set of two OPAs, to study charge photogeneration and transport in conjugated polymers and their donor-acceptor blends. (special issue devoted to the 80th birthday of S.A. Akhmanov)

  14. Majorana fermion fingerprints in spin-polarised scanning tunnelling microscopy

    NASA Astrophysics Data System (ADS)

    Kotetes, Panagiotis; Mendler, Daniel; Heimes, Andreas; Schön, Gerd

    2015-11-01

    We calculate the spatially resolved tunnelling conductance of topological superconductors (TSCs) based on ferromagnetic chains, measured by means of spin-polarised scanning tunnelling microscopy (SPSTM). Our analysis reveals novel signatures of MFs arising from the interplay of their strongly anisotropic spin-polarisation and the magnetisation content of the tip. We focus on the deep Yu-Shiba-Rusinov (YSR) limit where only YSR bound states localised in the vicinity of the adatoms govern the low-energy as also the topological properties of the system. Under these conditions, we investigate the occurrence of zero/finite bias peaks (ZBPs/FBPs) for a single or two coupled TSC chains forming a Josephson junction. Each TSC can host up to two Majorana fermions (MFs) per edge if chiral symmetry is preserved. Here we retrieve the conductance for all the accessible configurations of the MF number of each chain. Our results illustrate innovative spin-polarisation-sensitive experimental routes for arresting the MFs by either restoring or splitting the ZBP in a predictable fashion via: (i) weakly breaking chiral symmetry, e.g. by the SPSTM tip itself or by an external Zeeman field and (ii) tuning the superconducting phase difference of the TSCs, which is encoded in the 4π-Josephson coupling of neighbouring MFs.

  15. PoGOLite measurement of Crab polarisation and future plans

    NASA Astrophysics Data System (ADS)

    Pearce, Mark

    2016-07-01

    (For the PoGOLite Collaboration) The PoGOLite Pathfinder is a balloon-borne hard X-ray polarimeter designed for the observation of bright, ~1 Crab, sources. Polarisation is determined by measuring the azimuthal Compton scattering angle of incident X-rays in an array of plastic scintillators housed in an BGO anticoincidence well. The PoGOLite Pathfinder was launched from the SSC Esrange Space Centre in July 2013 resulting in a near-circumpolar flight of two weeks duration. The linear polarisation of hard X-ray emissions from the Crab was measured in a previously unexplored energy interval, 20-120 keV. The polarimetric response was characterised prior to flight using both polarised and unpolarised calibration sources. Systematic effects were addressed through observations of a background field. An upgraded polarimeter, PoGO+, is scheduled to fly in summer 2016 from Esrange. Results from the 2013 Pathfinder flight and prospects for the 2016 flight will be discussed.

  16. Characterization of Tetratricopeptide Repeat-Containing Proteins Critical for Cilia Formation and Function

    PubMed Central

    Xu, Yanan; Cao, Jingli; Huang, Shan; Feng, Di; Zhang, Wei; Zhu, Xueliang; Yan, Xiumin

    2015-01-01

    Cilia formation and function require a special set of trafficking machinery termed intraflagellar transport (IFT), consisting mainly of protein complexes IFT-A, IFT-B, BBSome, and microtubule-dependent molecular motors. Tetratricopeptide repeat-containing (TTC) proteins are widely involved in protein complex formation. Nine of them are known to serve as components of the IFT or BBSome complexes. How many TTC proteins are cilia-related and how they function, however, remain unclear. Here we show that twenty TTC genes were upregulated by at least 2-fold during the differentiation of cultured mouse tracheal epithelial cells (MTECs) into multiciliated cells. Our systematic screen in zebrafish identified four novel TTC genes, ttc4, -9c, -36, and -39c, that are critical for cilia formation and motility. Accordingly, their zebrafish morphants displayed typical ciliopathy-related phenotypes, including curved body, abnormal otolith, hydrocephalus, and defective left-right patterning. The morphants of ttc4 and ttc25, a known cilia-related gene, additionally showed pronephric cyst formation. Immunoprecipitation indicated associations of TTC4, -9c, -25, -36, and -39c with components or entire complexes of IFT-A, IFT-B, or BBSome, implying their participations in IFT or IFT-related activities. Our results provide a global view for the relationship between TTC proteins and cilia. PMID:25860617

  17. Flow induced by ependymal cilia dominates near-wall cerebrospinal fluid dynamics in the lateral ventricles

    PubMed Central

    Siyahhan, Bercan; Knobloch, Verena; de Zélicourt, Diane; Asgari, Mahdi; Schmid Daners, Marianne; Poulikakos, Dimos; Kurtcuoglu, Vartan

    2014-01-01

    While there is growing experimental evidence that cerebrospinal fluid (CSF) flow induced by the beating of ependymal cilia is an important factor for neuronal guidance, the respective contribution of vascular pulsation-driven macroscale oscillatory CSF flow remains unclear. This work uses computational fluid dynamics to elucidate the interplay between macroscale and cilia-induced CSF flows and their relative impact on near-wall dynamics. Physiological macroscale CSF dynamics are simulated in the ventricular space using subject-specific anatomy, wall motion and choroid plexus pulsations derived from magnetic resonance imaging. Near-wall flow is quantified in two subdomains selected from the right lateral ventricle, for which dynamic boundary conditions are extracted from the macroscale simulations. When cilia are neglected, CSF pulsation leads to periodic flow reversals along the ventricular surface, resulting in close to zero time-averaged force on the ventricle wall. The cilia promote more aligned wall shear stresses that are on average two orders of magnitude larger compared with those produced by macroscopic pulsatile flow. These findings indicate that CSF flow-mediated neuronal guidance is likely to be dominated by the action of the ependymal cilia in the lateral ventricles, whereas CSF dynamics in the centre regions of the ventricles is driven predominantly by wall motion and choroid plexus pulsation. PMID:24621815

  18. Dynamics of self-oscillating cilia designed from active polymer gels

    NASA Astrophysics Data System (ADS)

    Dayal, Pratyush; Bhattacharya, Amitabh; Kuksenok, Olga; Balazs, Anna C.

    2012-02-01

    Using theory and simulations, we design active synthetic surfaces which are capable of replicating functionalities of biological cilia. In order to design such exquisite biomimetic systems we harness unique properties of polymer gels that undergo photosensitive Belousov-Zhabotinsky (BZ) reaction. Powered by internalized BZ reaction these polymer gels swell and de-swell autonomously by chemo-mechanical transduction and therefore are ideal materials for designing our system. In order to simulate the dynamics of the BZ cilia in surrounding fluid we have developed a nonlinear hybrid 3D model which captures elasto-dynamics of polymer gel and diffusive exchange of BZ reagents between the gel and the fluid. Here we show that the geometrical arrangement of cilia and the distribution of BZ activator in the fluid determine the dynamic response of the cilia. We further show that using light as an external stimulus we can sequentially modulate height of individual cilium and thereby create the ``piano effect''. Finally, we demonstrate that synchronized oscillations in the cilia result from the distribution of BZ-activator in the surrounding fluid. Our findings can be used to design active surfaces which can be remotely tuned depending upon the magnitude of external stimuli.

  19. Proximal tubule proliferation is insufficient to induce rapid cyst formation after cilia disruption.

    PubMed

    Sharma, Neeraj; Malarkey, Erik B; Berbari, Nicolas F; O'Connor, Amber K; Vanden Heuvel, Gregory B; Mrug, Michal; Yoder, Bradley K

    2013-02-01

    Disrupting the function of cilia in mouse kidneys results in rapid or slow progression of cystic disease depending on whether the animals are juveniles or adults, respectively. Renal injury can also markedly accelerate the renal cyst formation that occurs after disruption of cilia in adult mice. Rates of cell proliferation are markedly higher in juvenile than adult kidneys and increase after renal injury, suggesting that cell proliferation may enhance the development of cysts. Here, we induced cilia loss in the kidneys of adult mice in the presence or absence of a Cux-1 transgene, which maintains cell proliferation. By using this model, we were able to avoid additional factors such as inflammation and dedifferentiation, which associate with renal injury and may also influence the rate of cystogenesis. After induction of cilia loss, cystic disease was not more pronounced in adult mice with the Cux-1 transgene compared with those without the transgene. In conclusion, these data suggest that proliferation is unlikely to be the sole mechanism underlying the rapid cystogenesis observed after injury in mice that lose cilia function in adulthood. PMID:23411784

  20. Simulation by using the lattice Boltzmann method of microscopic particle motion induced by artificial cilia

    NASA Astrophysics Data System (ADS)

    Alapati, Suresh; Che, Woo Seong; Mannoor, Madhusoodanan; Suh, Yong Kweon

    2016-06-01

    In this paper, we present the results obtained from the simulation of particle motion induced by the fluid flow driven by an array of beating artificial cilia inside a micro-channel. A worm-like-chain model is used to simulate the elastic cilia, and the lattice Boltzmann equation is used to compute the fluid flow. We employ a harmonic force at the extreme tip of each cilium to actuate it. Our simulation methods are first validated by applying them to the motion of a single cilium and a freely falling sphere. After validation, we simulate the fluid flow generated by an array of beating cilia and find that a maximum flow rate is achieved at an optimum sperm number. Next, we simulate the motion of a neutrally buoyant spherical particle at this optimum sperm number by tracking the particle motion with a smoothed profile method. We address the effect of the following parameters on the particle velocity: the gap between cilia and particle, the particle size, the cilia density, and the presence of an array of intermediate particles.

  1. A Group of ent-Kaurane Diterpenoids Inhibit Hedgehog Signaling and Induce Cilia Elongation

    PubMed Central

    Jiang, Shiyou; Du, Jiacheng; Kong, Qinghua; Li, Chaocui; Li, Yan; Sun, Handong; Pu, Jianxin; Mao, Bingyu

    2015-01-01

    The Hedgehog (Hh) signaling pathway plays important roles in the tumorigenesis of multiple cancers and is a key target for drug discovery. In a screen of natural products extracted from Chinese herbs, we identified eight ent-Kaurane diterpenoids and two triterpene dilactones as novel Hh pathway antagonists. Epistatic analyses suggest that these compounds likely act at the level or downstream of Smoothened (Smo) and upstream of Suppressor of Fused (Sufu). The ent-Kauranoid-treated cells showed elongated cilia, suppressed Smo trafficking to cilia, and mitotic defects, while the triterpene dilactones had no effect on the cilia and ciliary Smo. These ent-Kaurane diterpenoids provide new prototypes of Hh inhibitors, and are valuable probes for deciphering the mechanisms of Smo ciliary transport and ciliogenesis. PMID:26439749

  2. Ex vivo method for high resolution imaging of cilia motility in rodent airway epithelia.

    PubMed

    Francis, Richard; Lo, Cecilia

    2013-01-01

    An ex vivo technique for imaging mouse airway epithelia for quantitative analysis of motile cilia function important for insight into mucociliary clearance function has been established. Freshly harvested mouse trachea is cut longitudinally through the trachealis muscle and mounted in a shallow walled chamber on a glass-bottomed dish. The trachea sample is positioned along its long axis to take advantage of the trachealis muscle to curl longitudinally. This allows imaging of ciliary motion in the profile view along the entire tracheal length. Videos at 200 frames/sec are obtained using differential interference contrast microscopy and a high speed digital camera to allow quantitative analysis of cilia beat frequency and ciliary waveform. With the addition of fluorescent beads during imaging, cilia generated fluid flow also can be determined. The protocol time spans approximately 30 min, with 5 min for chamber preparation, 5-10 min for sample mounting, and 10-15 min for videomicroscopy. PMID:23963287

  3. 3D structure of eukaryotic flagella/cilia by cryo-electron tomography

    PubMed Central

    Ishikawa, Takashi

    2013-01-01

    Flagella/cilia are motile organelles with more than 400 proteins. To understand the mechanism of such complex systems, we need methods to describe molecular arrange-ments and conformations three-dimensionally in vivo. Cryo-electron tomography enabled us such a 3D structural analysis. Our group has been working on 3D structure of flagella/cilia using this method and revealed highly ordered and beautifully organized molecular arrangement. 3D structure gave us insights into the mechanism to gener-ate bending motion with well defined waveforms. In this review, I summarize our recent structural studies on fla-gella/cilia by cryo-electron tomography, mainly focusing on dynein microtubule-based ATPase motor proteins and the radial spoke, a regulatory protein complex. PMID:27493552

  4. Cycling of the signaling protein phospholipase D through cilia requires the BBSome only for the export phase

    PubMed Central

    Brown, Jason M.; Sampaio, Julio L.; Craft, Julie M.; Shevchenko, Andrej; Evans, James E.; Witman, George B.

    2013-01-01

    The BBSome is a complex of seven proteins, including BBS4, that is cycled through cilia by intraflagellar transport (IFT). Previous work has shown that the membrane-associated signaling protein phospholipase D (PLD) accumulates abnormally in cilia of Chlamydomonas reinhardtii bbs mutants. Here we show that PLD is a component of wild-type cilia but is enriched ∼150-fold in bbs4 cilia; this accumulation occurs progressively over time and results in altered ciliary lipid composition. When wild-type BBSomes were introduced into bbs cells, PLD was rapidly removed from the mutant cilia, indicating the presence of an efficient BBSome-dependent mechanism for exporting ciliary PLD. This export requires retrograde IFT. Importantly, entry of PLD into cilia is BBSome and IFT independent. Therefore, the BBSome is required only for the export phase of a process that continuously cycles PLD through cilia. Another protein, carbonic anhydrase 6, is initially imported normally into bbs4 cilia but lost with time, suggesting that its loss is a secondary effect of BBSome deficiency. PMID:23589493

  5. High e-vector acuity in the polarisation vision system of the fiddler crab Uca vomeris.

    PubMed

    How, Martin J; Pignatelli, Vincenzo; Temple, Shelby E; Marshall, N Justin; Hemmi, Jan M

    2012-06-15

    Polarisation vision is used by a variety of species in many important tasks, including navigation and orientation (e.g. desert ant), communication and signalling (e.g. stomatopod crustaceans), and as a possible substitute for colour vision (e.g. cephalopod molluscs). Fiddler crabs are thought to possess the anatomical structures necessary to detect polarised light, and occupy environments rich in polarisation cues. Yet little is known about the capabilities of their polarisation sense. A modified polarisation-only liquid crystal display and a spherical rotating treadmill were combined to test the responses of fiddler crabs to moving polarisation stimuli. The species Uca vomeris was found to be highly sensitive to polarised light and detected stimuli differing in e-vector angle by as little as 3.2 deg. This represents the most acute behavioural sensitivity to polarised light yet measured for a crustacean. The occurrence of null points in their discrimination curve indicates that this species employs an orthogonal (horizontal/vertical) receptor array for the detection of polarised light. PMID:22623201

  6. Mucociliary and long-term particle clearance in airways of patients with immotile cilia

    PubMed Central

    Möller, Winfried; Häußinger, Karl; Ziegler-Heitbrock, Löms; Heyder, Joachim

    2006-01-01

    Spherical monodisperse ferromagnetic iron oxide particles of 1.9 μm geometric and 4.2 μm aerodynamic diameter were inhaled by seven patients with primary ciliary dyskinesia (PCD) using the shallow bolus technique, and compared to 13 healthy non-smokers (NS) from a previous study. The bolus penetration front depth was limiting to the phase1 dead space volume. In PCD patients deposition was 58+/-8 % after 8 s breath holding time. Particle retention was measured by the magnetopneumographic method over a period of nine months. Particle clearance from the airways showed a fast and a slow phase. In PCD patients airway clearance was retarded and prolonged, 42+/-12 % followed the fast phase with a mean half time of 16.8+/-8.6 hours. The remaining fraction was cleared slowly with a half time of 121+/-25 days. In healthy NS 49+/-9 % of particles were cleared in the fast phase with a mean half time of 3.0+/-1.6 hours, characteristic of an intact mucociliary clearance. There was no difference in the slow clearance phase between PCD patients and healthy NS. Despite non-functioning cilia the effectiveness of airway clearance in PCD patients is comparable to healthy NS, with a prolonged kinetics of one week, which may primarily reflect the effectiveness of cough clearance. This prolonged airway clearance allows longer residence times of bacteria and viruses in the airways and may be one reason for increased frequency of infections in PCD patients. PMID:16423294

  7. Metachronal beating of cilia under the influence of Casson fluid and magnetic field

    NASA Astrophysics Data System (ADS)

    Akbar, Noreen Sher; Khan, Zafar Hayat

    2015-03-01

    Metachronal beating of cilia under the influence of Casson fluid and magnetic field is considered. The model for cilia literature is modelled for the first time. The governing coupled equations are constructed under long wavelength and low Reynold's number approximation. Exact solutions are evaluated for stream function and pressure gradient. The important results in this study are the variation of the Hartmann number M, Casson fluid parameter ζ. The velocity field increases due to the increase in Hartmann number M near the channel walls while velocity field decreases at the center of the channel. Comparative study is also made for Casson fluid with Newtonian fluid.

  8. Immotile cilia syndrome: A recombinant family at HLA-linked gene locus

    SciTech Connect

    Gasparini, P.; Grifa, A.; Oggiano, N.; Fabbrizzi, E.; Giorgi, P.L.

    1994-02-15

    The immotile-cilia syndrome (ICS) is an autosomal recessive trait of congenital dismobility or even complete immobility of cilia in the ciliated epithelia (MIM 244400). Recurrent upper respiratory infections in early childhood are the most common clinical findings. Recently a disease locus was mapped by sib pair analysis in two unrelated families on 6p tightly linked to HLA class II loci, such as DR and DQ. In order to confirm this assignment and to test the presence of possible heterogeneity, the authors analyzed several ICS families utilizing DNA makers of HLA class II region. Here they report the identification of a recombinant family at this locus. 3 refs., 1 fig.

  9. A missense mutation in DCDC2 causes human recessive deafness DFNB66, likely by interfering with sensory hair cell and supporting cell cilia length regulation

    PubMed Central

    Grati, M'hamed; Chakchouk, Imen; Ma, Qi; Bensaid, Mariem; Desmidt, Alexandra; Turki, Nouha; Yan, Denise; Baanannou, Aissette; Mittal, Rahul; Driss, Nabil; Blanton, Susan; Farooq, Amjad; Lu, Zhongmin; Liu, Xue Zhong; Masmoudi, Saber

    2015-01-01

    Hearing loss is the most common sensory deficit in humans. We show that a point mutation in DCDC2 (DCDC2a), a member of doublecortin domain-containing protein superfamily, causes non-syndromic recessive deafness DFNB66 in a Tunisian family. Using immunofluorescence on rat inner ear neuroepithelia, DCDC2a was found to localize to the kinocilia of sensory hair cells and the primary cilia of nonsensory supporting cells. DCDC2a fluorescence is distributed along the length of the kinocilium with increased density toward the tip. DCDC2a-GFP overexpression in non-polarized COS7 cells induces the formation of long microtubule-based cytosolic cables suggesting a role in microtubule formation and stabilization. Deafness mutant DCDC2a expression in hair cells and supporting cells causes cilium structural defects, such as cilium branching, and up to a 3-fold increase in length ratios. In zebrafish, the ortholog dcdc2b was found to be essential for hair cell development, survival and function. Our results reveal DCDC2a to be a deafness gene and a player in hair cell kinocilia and supporting cell primary cilia length regulation likely via its role in microtubule formation and stabilization. PMID:25601850

  10. Innervation of Gill Lateral Cells in the Bivalve Mollusc Crassostrea virginica Affects Cellular Membrane Potential and Cilia Activity

    PubMed Central

    Catapane, Edward J; Nelson, Michael; Adams, Trevon; Carroll, Margaret A

    2016-01-01

    Gill lateral cells of Crassostrea virginica are innervated by the branchial nerve, which contains serotonergic and dopaminergic fibers that regulate cilia beating rate. Terminal release of serotonin or dopamine results in an increase or decrease, respectively, of cilia beating rate in lateral gill cells. In this study we used the voltage sensitive fluorescent probe DiBAC4(3) to quantify changes in gill lateral cell membrane potential in response to electrical stimulation of the branchial nerve or to applications of serotonin and dopamine, and correlate these changes to cilia beating rates. Application of serotonin to gill lateral cells caused prolonged membrane depolarization, similar to plateau potentials, while increasing cilia beating rate. Application of dopamine hyperpolarized the resting membrane while decreasing cilia beating rate. Low frequency (5 Hz) electrical stimulations of the branchial nerve, which cause terminal release of endogenous serotonin, or high frequency (20 Hz) stimulations, which cause terminal release of endogenous dopamine, had the same effects on gill lateral cell membrane potentials and cilia beating rate as the respective applications of serotonin or dopamine. The study shows that innervation of gill lateral cells by the branchial nerve affects membrane potential as well as cilia beating rate, and demonstrates a strong correlation between changes in membrane potential and regulation of cilia beating rate. The study furthers the understanding of serotonin and dopamine signaling in the innervation and regulation of gill cilia in bivalves. The study also shows that voltage sensitive fluorescent probes like DiBAC 4(3) can be successfully used as an alternative to microelectrodes to measure changes in membrane potential of ciliated gill cells and other small cells with fast moving cilia. PMID:27489887

  11. The role of hair cells, cilia and ciliary motility in otolith formation in the zebrafish otic vesicle.

    PubMed

    Stooke-Vaughan, Georgina A; Huang, Peng; Hammond, Katherine L; Schier, Alexander F; Whitfield, Tanya T

    2012-05-01

    Otoliths are biomineralised structures required for the sensation of gravity, linear acceleration and sound in the zebrafish ear. Otolith precursor particles, initially distributed throughout the otic vesicle lumen, become tethered to the tips of hair cell kinocilia (tether cilia) at the otic vesicle poles, forming two otoliths. We have used high-speed video microscopy to investigate the role of cilia and ciliary motility in otolith formation. In wild-type ears, groups of motile cilia are present at the otic vesicle poles, surrounding the immotile tether cilia. A few motile cilia are also found on the medial wall, but most cilia (92-98%) in the otic vesicle are immotile. In mutants with defective cilia (iguana) or ciliary motility (lrrc50), otoliths are frequently ectopic, untethered or fused. Nevertheless, neither cilia nor ciliary motility are absolutely required for otolith tethering: a mutant that lacks cilia completely (MZovl) is still capable of tethering otoliths at the otic vesicle poles. In embryos with attenuated Notch signalling [mindbomb mutant or Su(H) morphant], supernumerary hair cells develop and otolith precursor particles bind to the tips of all kinocilia, or bind directly to the hair cells' apical surface if cilia are absent [MZovl injected with a Su(H)1+2 morpholino]. However, if the first hair cells are missing (atoh1b morphant), otolith formation is severely disrupted and delayed. Our data support a model in which hair cells produce an otolith precursor-binding factor, normally localised to tether cell kinocilia. We also show that embryonic movement plays a minor role in the formation of normal otoliths. PMID:22461562

  12. Constructing and deconstructing roles for the primary cilium in tissue architecture and cancer

    PubMed Central

    Seeley, E. Scott; Nachury, Maxence V.

    2010-01-01

    Primary cilia are exquisitely designed sensory machines that have evolved at least three distinct sensory modalities to monitor the extracellular environment. The presence and activation of growth factor, morphogen, and hormone receptors within the confines of the ciliary membrane, the intrinsic physical relationship between the ciliary axoneme and the centriole, and the preferential assembly of primary cilia on the apical surfaces of tissue epithelia highlight the importance of this organelle in the establishment and maintenance of tissue architecture and homeostasis. Accordingly, recent studies begin to suggest roles for these organelles in oncogenesis and tumor suppression. Here, we review the sensory properties of primary cilia, assess the “history” of the primary cilium in cancer, and draw upon recent findings in a discussion of how the primary cilium may influence tissue architecture and neoplasia. PMID:20362097

  13. Primary Ciliary Dyskinesia.

    PubMed

    Knowles, Michael R; Zariwala, Maimoona; Leigh, Margaret

    2016-09-01

    Primary ciliary dyskinesia (PCD) is a recessive genetically heterogeneous disorder of motile cilia with chronic otosinopulmonary disease and organ laterality defects in ∼50% of cases. The prevalence of PCD is difficult to determine. Recent diagnostic advances through measurement of nasal nitric oxide and genetic testing has allowed rigorous diagnoses and determination of a robust clinical phenotype, which includes neonatal respiratory distress, daily nasal congestion, and wet cough starting early in life, along with organ laterality defects. There is early onset of lung disease in PCD with abnormal airflow mechanics and radiographic abnormalities detected in infancy and early childhood. PMID:27514592

  14. Intestinal cell kinase, a protein associated with endocrine-cerebro-osteodysplasia syndrome, is a key regulator of cilia length and Hedgehog signaling

    PubMed Central

    Moon, Heejung; Song, Jieun; Shin, Jeong-Oh; Lee, Hankyu; Kim, Hong-Kyung; Eggenschwiller, Jonathan T.; Bok, Jinwoong; Ko, Hyuk Wan

    2014-01-01

    Endocrine-cerebro-osteodysplasia (ECO) syndrome is a recessive genetic disorder associated with multiple congenital defects in endocrine, cerebral, and skeletal systems that is caused by a missense mutation in the mitogen-activated protein kinase-like intestinal cell kinase (ICK) gene. In algae and invertebrates, ICK homologs are involved in flagellar formation and ciliogenesis, respectively. However, it is not clear whether this role of ICK is conserved in mammals and how a lack of functional ICK results in the characteristic phenotypes of human ECO syndrome. Here, we generated Ick knockout mice to elucidate the precise role of ICK in mammalian development and to examine the pathological mechanisms of ECO syndrome. Ick null mouse embryos displayed cleft palate, hydrocephalus, polydactyly, and delayed skeletal development, closely resembling ECO syndrome phenotypes. In cultured cells, down-regulation of Ick or overexpression of kinase-dead or ECO syndrome mutant ICK resulted in an elongation of primary cilia and abnormal Sonic hedgehog (Shh) signaling. Wild-type ICK proteins were generally localized in the proximal region of cilia near the basal bodies, whereas kinase-dead ICK mutant proteins accumulated in the distal part of bulged ciliary tips. Consistent with these observations in cultured cells, Ick knockout mouse embryos displayed elongated cilia and reduced Shh signaling during limb digit patterning. Taken together, these results indicate that ICK plays a crucial role in controlling ciliary length and that ciliary defects caused by a lack of functional ICK leads to abnormal Shh signaling, resulting in congenital disorders such as ECO syndrome. PMID:24853502

  15. Surface Parameter Estimation using Interferometric Coherences between Different Polarisations

    NASA Astrophysics Data System (ADS)

    Hajnsek, I.; Alvarez-Perez, J.-L.; Papathanassiou, K. P.; Moreira, A.; Cloude, S. R.

    2003-04-01

    In this work the potential of using the interferometric coherence at different polarisations over surface scat- terers in order to extract information about surface parameters is investigated. For the first time the sensitivity of the indi- vidual coherence contributions to surface roughness and moisture conditions is discussed and simulated using a novel hy- brid polarimetric surface scattering model. The model itself consists of two components, a coherent part obtained from the extended Bragg model and an incoherent part obtained from the integral equation model. Finally, experimental airborne SAR data are used to validate the modeled elements of the Pauli scattering vector.

  16. Direct evidence for BBSome-associated intraflagellar transport reveals distinct properties of native mammalian cilia

    PubMed Central

    Williams, Corey L.; McIntyre, Jeremy C.; Norris, Stephen R.; Jenkins, Paul M.; Zhang, Lian; Pei, Qinglin; Verhey, Kristen; Martens, Jeffrey R.

    2014-01-01

    Cilia dysfunction underlies a class of human diseases with variable penetrance in different organ systems. Across eukaryotes, intraflagellar transport (IFT) facilitates cilia biogenesis and cargo trafficking, but our understanding of mammalian IFT is insufficient. Here we perform live analysis of cilia ultrastructure, composition and cargo transport in native mammalian tissue using olfactory sensory neurons. Proximal and distal axonemes of these neurons show no bias towards IFT kinesin-2 choice, and Kif17 homodimer is dispensable for distal segment IFT. We identify Bardet–Biedl syndrome proteins (BBSome) as bona fide constituents of IFT in olfactory sensory neurons, and show that they exist in 1:1 stoichiometry with IFT particles. Conversely, subpopulations of peripheral membrane proteins, as well as transmembrane olfactory signalling pathway components, are capable of IFT but with significantly less frequency and/or duration. Our results yield a model for IFT and cargo trafficking in native mammalian cilia and may explain the penetrance of specific ciliopathy phenotypes in olfactory neurons. PMID:25504142

  17. Exact solution of cilia induced flow of a Jeffrey fluid in an inclined tube.

    PubMed

    Maqbool, K; Shaheen, S; Mann, A B

    2016-01-01

    The present study investigated the cilia induced flow of MHD Jeffrey fluid through an inclined tube. This study is carried out under the assumptions of long wavelength and low Reynolds number approximations. Exact solutions for the velocity profile, pressure rise, pressure gradient, volume flow rate and stream function are obtained. Effects of pertinent physical parameters on the computational results are presented graphically. PMID:27610298

  18. Levonorgestrel decreases cilia beat frequency of human fallopian tubes and rat oviducts without changing morphological structure.

    PubMed

    Zhao, Weihong; Zhu, Qian; Yan, Mingxing; Li, Cheng; Yuan, Jiangjing; Qin, Guojuan; Zhang, Jian

    2015-02-01

    Levonorgestrel, a derivative of progesterone, effectively protects women against unwanted pregnancy as an emergency contraceptive. Previous studies have not been successful in determining the mechanism by which levonorgestrel acts. In the present study we analysed cilia beat action and cilia morphology following levonorgestrel exposure in vitro and in vivo using both light and electron microscopy. There was a significant decrease in the ciliary beat frequency (CBF) of human fallopian tubes between mucosal explants bathed in 5 μmol/L levonorgestrel and those bathed in medium alone (P < 0.05). There was a tendency for CBF to decrease more in the ampulla than in isthmus, but there were no differences between the proliferative and secretory phases. In rat oviducts, levonorgestrel produced a similar reduction in CBF (~ 10%) compared with the saline control group (P < 0.05). Histological and ultrastructural analysis demonstrated no changes in the percentage of ciliated cells or in the classic '9 + 2' structure of cilia following levonorgestrel treatment in either system. Thus, levonorgestrel reduces CBF without damaging cilia morphology. Decreases in CBF may indicate a pathological role for levonorgestrel in the transportation of the ovum and zygote in the fallopian tube. PMID:25399777

  19. Dynamics of Individual cilia to external loading- A simple one dimensional picture

    NASA Astrophysics Data System (ADS)

    Swaminathan, Vinay; Hill, David; Superfine, R.

    2008-10-01

    From being called the cellular janitors to swinging debauchers, cilia have captured the fascinations of researchers for over 200 years. In cystic fibrosis and chronic obstructive pulmonary disease where the cilia loses it's function, the protective mucus layer in the lung thickens and mucociliary clearance breaks down, leading to inflammation along the airways and an increased rate of infection. The mechanistic understanding of mucus clearance depends on a quantitative assessment of the axoneme dynamics and the maximum force the cilia are capable of generating and imparting to the mucus layer. Similar to the situation in molecular motors, detailed quantitative measurements of dynamics under applied load conditions are expected to be essential in developing predictive models. Based on our measurements of the dynamics of individual ciliary motion in the human bronchial epithelial cell under the application of an applied load, we present a simple one dimensional model for the axoneme dynamics and quantify the axoneme stiffness, the internal force generated by the axoneme, the stall force and show how the dynamics sheds insight on the time dependence of the internal force generation. The internal force generated by the axoneme is related to the ability of cilia to propel fluids and to their potential role in force sensing.

  20. A Cilia Independent Role of Ift88/Polaris during Cell Migration

    PubMed Central

    Hamann, Christoph; Powelske, Christian; Mergen, Miriam; Herbst, Henriette; Kotsis, Fruzsina; Nitschke, Roland; Kuehn, E. Wolfgang

    2015-01-01

    Ift88 is a central component of the intraflagellar transport (Ift) complex B, essential for the building of cilia and flagella from single cell organisms to mammals. Loss of Ift88 results in the absence of cilia and causes left-right asymmetry defects, disordered Hedgehog signaling, and polycystic kidney disease, all of which are explained by aberrant ciliary function. In addition, a number of extraciliary functions of Ift88 have been described that affect the cell-cycle, mitosis, and targeting of the T-cell receptor to the immunological synapse. Similarly, another essential ciliary molecule, the kinesin-2 subunit Kif3a, which transports Ift-B in the cilium, affects microtubule (MT) dynamics at the leading edge of migrating cells independently of cilia. We now show that loss of Ift88 impairs cell migration irrespective of cilia. Ift88 is required for the polarization of migrating MDCK cells, and Ift88 depleted cells have fewer MTs at the leading edge. Neither MT dynamics nor MT nucleation are dependent on Ift88. Our findings dissociate the function of Ift88 from Kif3a outside the cilium and suggest a novel extraciliary function for Ift88. Future studies need to address what unifying mechanism underlies the different extraciliary functions of Ift88. PMID:26465598

  1. General and cell-type specific mechanisms target TRPP2/PKD-2 to cilia.

    PubMed

    Bae, Young-Kyung; Qin, Hongmin; Knobel, Karla M; Hu, Jinghua; Rosenbaum, Joel L; Barr, Maureen M

    2006-10-01

    Ciliary localization of the transient receptor potential polycystin 2 channel (TRPP2/PKD-2) is evolutionarily conserved, but how TRPP2 is targeted to cilia is not known. In this study, we characterize the motility and localization of PKD-2, a TRPP2 homolog, in C. elegans sensory neurons. We demonstrate that GFP-tagged PKD-2 moves bidirectionally in the dendritic compartment. Furthermore, we show a requirement for different molecules in regulating the ciliary localization of PKD-2. PKD-2 is directed to moving dendritic particles by the UNC-101/adaptor protein 1 (AP-1) complex. When expressed in non-native neurons, PKD-2 remains in cell bodies and is not observed in dendrites or cilia, indicating that cell-type specific factors are required for directing PKD-2 to the dendrite. PKD-2 stabilization in cilia and cell bodies requires LOV-1, a functional partner and a TRPP1 homolog. In lov-1 mutants, PKD-2 is greatly reduced in cilia and forms abnormal aggregates in neuronal cell bodies. Intraflagellar transport (IFT) is not essential for PKD-2 dendritic motility or access to the cilium, but may regulate PKD-2 ciliary abundance. We propose that both general and cell-type-specific factors govern TRPP2/PKD-2 subcellular distribution by forming at least two steps involving somatodendritic and ciliary sorting decisions. PMID:16943275

  2. Distinctive features of cilia in metazoans and their significance for systematics.

    PubMed

    Tyler, S

    1979-01-01

    A comparative study of epidermal cilia in the Turbellaria and Nemertea has revealed features in these organelles that are specific to certain taxonomic groups. Turbellarians of the order Acoela, in particular, have a characteristic pattern of axonemal filament termination in the distal tips of their cilia and a characteristic ciliary rootlet system that is not seen in other turbellarian orders nor in other metazoans. Each epidermal cilium in acoels has a typical 9 + 2 axonemal pattern through the main part of its length, but near its distal tip there is an abrupt shelf-life narrowing at which filaments 4-7 terminate; filaments 1, 2, 8 and 9 continue into the thinner distal-most part of the shaft along with singlet microtubules from the axonemal center. The rootlet system in acoel cilia involves an interconnecting pattern with lateral connectives. The unique structure of these cilia has systematic and phylogenetic significance for the Acoela, and it is argued that ultrastructural characters in general, including characters of organelles, can be validly applied to the phylogeny and systematics of the Metazoa. PMID:494232

  3. Prickle3 synergizes with Wtip to regulate basal body organization and cilia growth

    PubMed Central

    Chu, Chih-Wen; Ossipova, Olga; Ioannou, Andriani; Sokol, Sergei Y.

    2016-01-01

    PCP proteins maintain planar polarity in many epithelial tissues and have been implicated in cilia development in vertebrate embryos. In this study we examine Prickle3 (Pk3), a vertebrate homologue of Drosophila Prickle, in Xenopus gastrocoel roof plate (GRP). GRP is a tissue equivalent to the mouse node, in which cilia-generated flow promotes left-right patterning. We show that Pk3 is enriched at the basal body of GRP cells but is recruited by Vangl2 to anterior cell borders. Interference with Pk3 function disrupted the anterior polarization of endogenous Vangl2 and the posterior localization of cilia in GRP cells, demonstrating its role in PCP. Strikingly, in cells with reduced Pk3 activity, cilia growth was inhibited and γ-tubulin and Nedd1 no longer associated with the basal body, suggesting that Pk3 has a novel function in basal body organization. Mechanistically, this function of Pk3 may involve Wilms tumor protein 1-interacting protein (Wtip), which physically associates with and cooperates with Pk3 to regulate ciliogenesis. We propose that, in addition to cell polarity, PCP components control basal body organization and function. PMID:27062996

  4. Loss of Dishevelleds disrupts planar polarity in ependymal motile cilia and results in hydrocephalus

    PubMed Central

    Ohata, Shinya; Nakatani, Jin; Herranz-Pérez, Vicente; Cheng, JrGang; Belinson, Haim; Inubushi, Toshiro; Snider, William D.; García-Verdugo, Jose Manuel; Wynshaw-Boris, Anthony; Álvarez-Buylla, Arturo

    2014-01-01

    SUMMARY Defects in ependymal (E) cells, which line the ventricle and generate cerebrospinal fluid flow through ciliary beating, can cause hydrocephalus. Dishevelled genes (Dvls) are essential for Wnt signaling and Dvl2 has been shown to localize to the rootlet of motile cilia. Using the hGFAP-Cre;Dvl1−/−;2flox/flox;3+/− mouse, we show that compound genetic ablation of Dvls causes hydrocephalus. In hGFAP-Cre;Dvl1−/−;2flox/flox;3+/− mutants, E cells differentiated normally, but the intracellular and intercellular rotational alignments of ependymal motile cilia were disrupted. As a consequence, the fluid flow generated by the hGFAP-Cre;Dvl1−/−;2flox/flox;3+/− E cells was significantly slower than that observed in control mice. Dvls were also required for the proper positioning of motile cilia on the apical surface. Tamoxifen-induced conditional removal of Dvls in adult mice also resulted in defects in intracellular rotational alignment and positioning of ependymal motile cilia. These results suggest that Dvls are continuously required for E cell planar polarity and may prevent hydrocephalus. PMID:25043421

  5. Loss of Dishevelleds disrupts planar polarity in ependymal motile cilia and results in hydrocephalus.

    PubMed

    Ohata, Shinya; Nakatani, Jin; Herranz-Pérez, Vicente; Cheng, JrGang; Belinson, Haim; Inubushi, Toshiro; Snider, William D; García-Verdugo, Jose Manuel; Wynshaw-Boris, Anthony; Alvarez-Buylla, Arturo

    2014-08-01

    Defects in ependymal (E) cells, which line the ventricle and generate cerebrospinal fluid flow through ciliary beating, can cause hydrocephalus. Dishevelled genes (Dvls) are essential for Wnt signaling, and Dvl2 has been shown to localize to the rootlet of motile cilia. Using the hGFAP-Cre;Dvl1(-/-);2(flox/flox);3(+/-) mouse, we show that compound genetic ablation of Dvls causes hydrocephalus. In hGFAP-Cre;Dvl1(-/-);2(flox/flox);3(+/-) mutants, E cells differentiated normally, but the intracellular and intercellular rotational alignments of ependymal motile cilia were disrupted. As a consequence, the fluid flow generated by the hGFAP-Cre;Dvl1(-/-);2(flox/flox);3(+/-) E cells was significantly slower than that observed in control mice. Dvls were also required for the proper positioning of motile cilia on the apical surface. Tamoxifen-induced conditional removal of Dvls in adult mice also resulted in defects in intracellular rotational alignment and positioning of ependymal motile cilia. These results suggest that Dvls are continuously required for E cell planar polarity and may prevent hydrocephalus. PMID:25043421

  6. Polarisation singularities in disordered photonic crystal waveguides for on-chip spin-photon entanglement

    NASA Astrophysics Data System (ADS)

    Beggs, Daryl; Lang, Ben; Young, Andrew; Oulton, Ruth

    A polarisation singularity occurs at a position in a vector field where one of the parameters of the local polarisation ellipse (handedness, eccentricity or orientation) becomes singular. With the vector nature of electromagnetic fields, optics is an obvious place for the study of polarisation singularities, and they can be found in systems ranging from tightly focused beams to speckle fields. Here we demonstrate that photonic crystal waveguides support on-chip polarisation singularities. As Bloch waves, the eigenmodes of photonic crystal waveguides possess a strong longitudinal, as well as transverse, component of their electric field. The spatial dependence of both these components and the phase between them ensures a rich and complex polarisation landscape in the waveguide. Recently, the use of polarisation singularities found in photonic crystal waveguides is generating much interest for integrated quantum information applications, as they can couple the spin-states of electrons confined to quantum dots to the optical modes of the waveguide. For example, at a circular-point (C-point), the sign of the local helicity is governed by the propagation direction of the optical mode, which allows for spin-photon coupling to one direction only. However, any real system will inevitably contain imperfections, and it is not obvious that the polarisation singularities will persist in the disordered waveguides. Here, we use calculations of the eigenmodes of disordered waveguides to demonstrate that the polarisation singularities persist far beyond realistically expected levels of disorder.

  7. Compact prisms for polarisation splitting of fibre laser beams

    SciTech Connect

    Davydov, B L; Yagodkin, D I

    2005-11-30

    Simple compact monoprisms for spatial splitting of polarised laser beams with relatively small diameters (no more than 1 mm) are considered. Prisms can be made of optically inactive CaCO{sub 3}, {alpha}-BaB{sub 2}O{sub 4} ({alpha}-BBO), LiIO{sub 3}, LiNbO{sub 3}, YVO{sub 4}, and TiO{sub 2} crystals known in polarisation optics. The exact solution of the Snell equation for the extraordinary wave reflected from a surface arbitrarily tilted to its wave vector is obtained. The analysis of variants of the solution allows the fabrication of prisms with any deviation angles of the extraordinary wave by preserving the propagation direction of the ordinary wave. Three variants of prisms are considered: with minimised dimensions, with the Brewster output of the extraordinary beam, and with the deviation of the extraordinary wave by 90{sup 0}. Calcite prisms with the deviation angles for the extraordinary beam {approx}19{sup 0} and 90{sup 0} are tested experimentally. (control of laser radiation parameters)

  8. Tubulin tyrosine ligase-like genes ttll3 and ttll6 maintain zebrafish cilia structure and motility.

    PubMed

    Pathak, Narendra; Austin, Christina A; Drummond, Iain A

    2011-04-01

    Tubulin post-translational modifications generate microtubule heterogeneity and modulate microtubule function, and are catalyzed by tubulin tyrosine ligase-like (TTLL) proteins. Using antibodies specific to monoglycylated, polyglycylated, and glutamylated tubulin in whole mount immunostaining of zebrafish embryos, we observed distinct, tissue-specific patterns of tubulin modifications. Tubulin modification patterns in cilia correlated with the expression of ttll3 and ttll6 in ciliated cells. Expression screening of all zebrafish tubulin tyrosine ligase-like genes revealed additional tissue-specific expression of ttll1 in brain neurons, ttll4 in muscle, and ttll7 in otic placodes. Knockdown of ttll3 eliminated cilia tubulin glycylation but had surprisingly mild effects on cilia structure and motility. Similarly, knockdown of ttll6 strongly reduced cilia tubulin glutamylation but only partially affected cilia structure and motility. Combined loss of function of ttll3 and ttll6 caused near complete loss of cilia motility and induced a variety of axonemal ultrastructural defects similar to defects previously observed in zebrafish fleer mutants, which were shown to lack tubulin glutamylation. Consistently, we find that fleer mutants also lack tubulin glycylation. These results indicate that tubulin glycylation and glutamylation have overlapping functions in maintaining cilia structure and motility and that the fleer/dyf-1 TPR protein is required for both types of tubulin post-translational modification. PMID:21262966

  9. Sperm-Associated Antigen 6 (SPAG6) Deficiency and Defects in Ciliogenesis and Cilia Function: Polarity, Density, and Beat

    PubMed Central

    Teves, Maria E.; Sears, Patrick R.; Li, Wei; Zhang, Zhengang; Tang, Waixing; van Reesema, Lauren; Costanzo, Richard M.; Davis, C. William; Knowles, Michael R.; Strauss, Jerome F.; Zhang, Zhibing

    2014-01-01

    SPAG6, an axoneme central apparatus protein, is essential for function of ependymal cell cilia and sperm flagella. A significant number of Spag6-deficient mice die with hydrocephalus, and surviving males are sterile because of sperm motility defects. In further exploring the ciliary dysfunction in Spag6-null mice, we discovered that cilia beat frequency was significantly reduced in tracheal epithelial cells, and that the beat was not synchronized. There was also a significant reduction in cilia density in both brain ependymal and trachea epithelial cells, and cilia arrays were disorganized. The orientation of basal feet, which determines the direction of axoneme orientation, was apparently random in Spag6-deficient mice, and there were reduced numbers of basal feet, consistent with reduced cilia density. The polarized epithelial cell morphology and distribution of intracellular mucin, α-tubulin, and the planar cell polarity protein, Vangl2, were lost in Spag6-deficient tracheal epithelial cells. Polarized epithelial cell morphology and polarized distribution of α-tubulin in tracheal epithelial cells was observed in one-week old wild-type mice, but not in the Spag6-deficient mice of the same age. Thus, the cilia and polarity defects appear prior to 7 days post-partum. These findings suggest that SPAG6 not only regulates cilia/flagellar motility, but that in its absence, ciliogenesis, axoneme orientation, and tracheal epithelial cell polarity are altered. PMID:25333478

  10. Shot and Patronin polarise microtubules to direct membrane traffic and biogenesis of microvilli in epithelia.

    PubMed

    Khanal, Ichha; Elbediwy, Ahmed; Diaz de la Loza, Maria Del Carmen; Fletcher, Georgina C; Thompson, Barry J

    2016-07-01

    In epithelial tissues, polarisation of microtubules and actin microvilli occurs along the apical-basal axis of each cell, yet how these cytoskeletal polarisation events are coordinated remains unclear. Here, we examine the hierarchy of events during cytoskeletal polarisation in Drosophila melanogaster epithelia. Core apical-basal polarity determinants polarise the spectrin cytoskeleton to recruit the microtubule-binding proteins Patronin (CAMSAP1, CAMSAP2 and CAMPSAP3 in humans) and Shortstop [Shot; MACF1 and BPAG1 (also known as DST) in humans] to the apical membrane domain. Patronin and Shot then act to polarise microtubules along the apical-basal axis to enable apical transport of Rab11 endosomes by the Nuf-Dynein microtubule motor complex. Finally, Rab11 endosomes are transferred to the MyoV (also known as Didum in Drosophila) actin motor to deliver the key microvillar determinant Cadherin 99C to the apical membrane to organise the biogenesis of actin microvilli. PMID:27231092

  11. Shot and Patronin polarise microtubules to direct membrane traffic and biogenesis of microvilli in epithelia

    PubMed Central

    Khanal, Ichha; Elbediwy, Ahmed; Diaz de la Loza, Maria del Carmen; Fletcher, Georgina C.

    2016-01-01

    ABSTRACT In epithelial tissues, polarisation of microtubules and actin microvilli occurs along the apical-basal axis of each cell, yet how these cytoskeletal polarisation events are coordinated remains unclear. Here, we examine the hierarchy of events during cytoskeletal polarisation in Drosophila melanogaster epithelia. Core apical-basal polarity determinants polarise the spectrin cytoskeleton to recruit the microtubule-binding proteins Patronin (CAMSAP1, CAMSAP2 and CAMPSAP3 in humans) and Shortstop [Shot; MACF1 and BPAG1 (also known as DST) in humans] to the apical membrane domain. Patronin and Shot then act to polarise microtubules along the apical-basal axis to enable apical transport of Rab11 endosomes by the Nuf–Dynein microtubule motor complex. Finally, Rab11 endosomes are transferred to the MyoV (also known as Didum in Drosophila) actin motor to deliver the key microvillar determinant Cadherin 99C to the apical membrane to organise the biogenesis of actin microvilli. PMID:27231092

  12. Polarisation Dynamics of Vector Soliton Molecules in Mode Locked Fibre Laser

    PubMed Central

    Tsatourian, Veronika; Sergeyev, Sergey V.; Mou, Chengbo; Rozhin, Alex; Mikhailov, Vitaly; Rabin, Bryan; Westbrook, Paul S.; Turitsyn, Sergei K.

    2013-01-01

    Two fundamental laser physics phenomena - dissipative soliton and polarisation of light are recently merged to the concept of vector dissipative soliton (VDS), viz. train of short pulses with specific state of polarisation (SOP) and shape defined by an interplay between anisotropy, gain/loss, dispersion, and nonlinearity. Emergence of VDSs is both of the fundamental scientific interest and is also a promising technique for control of dynamic SOPs important for numerous applications from nano-optics to high capacity fibre optic communications. Using specially designed and developed fast polarimeter, we present here the first experimental results on SOP evolution of vector soliton molecules with periodic polarisation switching between two and three SOPs and superposition of polarisation switching with SOP precessing. The underlying physics presents an interplay between linear and circular birefringence of a laser cavity along with light induced anisotropy caused by polarisation hole burning. PMID:24193374

  13. Mechanisms of cilia-driven transport in the airways in the absence of mucus.

    PubMed

    Bermbach, Saskia; Weinhold, Karina; Roeder, Thomas; Petersen, Frank; Kugler, Christian; Goldmann, Torsten; Rupp, Jan; König, Peter

    2014-07-01

    Airway mucus is thought to be required for the clearance of inhaled particles by mucociliary transport, but this view has recently been challenged. To test if mucus is necessary for cilia-driven particle transport, we removed mucus from murine and human ex vivo airway preparations by thorough rinsing with buffer with or without additional dithiothreitol washing. The transport of particles with diameters of 4.5 μm, 200 nm, and 40 nm and of bacteria was analyzed by video microscopy. Complete removal of mucus was verified by wheat germ agglutinin staining and by scanning electron microscopy. In the absence of mucus, we observed efficient transport of particles and bacteria by direct cilia-mediated propulsion or via fluid flow generated by ciliary beating. Virus-sized particles had the tendency to attach to cilia. Because direct contact of particles with ciliated cells occurs in the absence of mucus, we examined if this direct interaction changes epithelial function. Neither bacteria- nor LPS-induced nuclear translocation of NF-κB p65 in ciliated cells occurred, indicating that mere contact between ciliated cells and bacteria during transport does not activate the epithelium. Attachment of virus-sized particles to cilia could induce mucus release and/or increase the ciliary beat frequency. Our results indicate that cilia-driven transport of particles with various sizes is possible in murine and human airways without the presence of mucus. If mucus-free transport fails, the epithelium can react by releasing mucus or increasing the ciliary beat frequency to maintain particle transport. PMID:24467665

  14. The primary cilium as a gravitational force transducer and a regulator of transcriptional noise.

    PubMed

    Moorman, Stephen J; Shorr, Ardon Z

    2008-08-01

    Circumstantial evidence has suggested that the primary cilium might function as a gravity sensor. Direct evidence of its gravity-sensing function has recently been provided by studies of rohon beard neurons. These neurons showed changes in the variability of gene expression levels that are linked to the cyclic changes in the Earth's gravitational field due to the Sun and Moon. These cyclic changes also cause the tides. Rohon beard neurons, after the primary cilia have been selectively destroyed, no longer show changes in gene expression variability linked to the cyclic changes in Earth's gravitational field. After the neurons regrow their primary cilia, the link between variability in gene expression levels and the Earth's changing gravitational field returns. This suggests two new functions for the primary cilia, detecting the cyclical changes in the Earth's gravitational field and transducing those changes into changes in the variability (stochastic nature) of gene expression. PMID:18366139

  15. Genetics and Biology of Primary Ciliary Dyskinesia

    PubMed Central

    Horani, Amjad; Ferkol, Thomas W; Dutcher, Susan K.; Brody, Steven L

    2016-01-01

    Summary Ciliopathies are a growing class of disorders caused by abnormal ciliary axonemal structure and function. Our understanding of the complex genetic and functional phenotypes of these conditions has rapidly progressed. Primary ciliary dyskinesia (PCD) remains the sole genetic disorder of motile cilia dysfunction. However, unlike many Mendelian genetic disorders, PCD is not caused by mutations in a single gene or locus, but rather, autosomal recessive mutation in one of many genes that lead to a similar phenotype. The first reported PCD mutations, more than a decade ago, identified genes encoding known structural components of the ciliary axoneme. In recent years, mutations in genes encoding novel cytoplasmic and regulatory proteins have been discovered. These findings have provided new insights into the functions of the motile cilia, and a better understanding of motile cilia disease. Advances in genetic tools will soon allow more precise genetic testing, mandating that clinicians must understand the genetic basis of PCD. Here, we review genetic mutations, their biological impact on cilia structure and function, and the implication of emerging genetic diagnostic tools. PMID:26476603

  16. Endocytic Control of Cellular Signaling at the Primary Cilium.

    PubMed

    Pedersen, Lotte B; Mogensen, Johanne B; Christensen, Søren T

    2016-09-01

    Primary cilia are dynamic signaling organelles that project from the cell surface to sense diverse chemical, physical and morphogenetic cues. Ciliary defects therefore cause diseases (ciliopathies) that affect multiple organs in developing and adult organisms. Cilia-mediated signaling involves the orchestrated movement of signaling proteins in and out of the ciliary compartment, including movement of receptors such as the Sonic Hedgehog (Shh) receptor Patched 1 (PTCH1), Smoothened (SMO), and various other G protein-coupled receptors (GPCRs), as well as transforming growth factor β (TGF-β) receptors I and II (TGF-β-RI/II). We provide here a current understanding of trafficking events associated with cilia-mediated signaling, with emphasis on the involvement of clathrin-dependent receptor-mediated endocytosis in regulating ciliary Shh and TGF-β signaling. PMID:27364476

  17. Phase delay of polarisation modes in elastically twisted spun fibres

    SciTech Connect

    Morshnev, Sergey K; Chamorovsky, Yury K; Vorob'ev, Igor' L

    2011-05-31

    The evolution of the phase delay between linearly polarised orthogonal modes in a spun fibre elastically twisted around its axis has been studied experimentally and theoretically using a model for a helical structure of the built-in linear birefringence axes. The phase delay is a sinusoidal function of elastic twist angle, with an amplitude and period dependent on fibre parameters: spin pitch and built-in linear birefringence beat length. It is shown that, at a known spin pitch, phase delay versus elastic twist angle data can be used to determine the beat length of built-in linear birefringence in the range 0.01 to 100 mm. The theoretical analysis results are supported by experimental data for conventional and microstructured spun fibres. (fibre optics)

  18. Measurement of CP asymmetries and polarisation fractions in decays

    NASA Astrophysics Data System (ADS)

    Aaij, R.; Adeva, B.; Adinolfi, M.; Affolder, A.; Ajaltouni, Z.; Akar, S.; Albrecht, J.; Alessio, F.; Alexander, M.; Ali, S.; Alkhazov, G.; Alvarez Cartelle, P.; Alves, A. A.; Amato, S.; Amerio, S.; Amhis, Y.; An, L.; Anderlini, L.; Anderson, J.; Andreassen, R.; Andreotti, M.; Andrews, J. E.; Appleby, R. B.; Aquines Gutierrez, O.; Archilli, F.; Artamonov, A.; Artuso, M.; Aslanides, E.; Auriemma, G.; Baalouch, M.; Bachmann, S.; Back, J. J.; Badalov, A.; Baesso, C.; Baldini, W.; Barlow, R. J.; Barschel, C.; Barsuk, S.; Barter, W.; Batozskaya, V.; Battista, V.; Bay, A.; Beaucourt, L.; Beddow, J.; Bedeschi, F.; Bediaga, I.; Bel, L. J.; Belogurov, S.; Belyaev, I.; Ben-Haim, E.; Bencivenni, G.; Benson, S.; Benton, J.; Berezhnoy, A.; Bernet, R.; Bertolin, A.; Bettler, M.-O.; van Beuzekom, M.; Bien, A.; Bifani, S.; Bird, T.; Bizzeti, A.; Blake, T.; Blanc, F.; Blouw, J.; Blusk, S.; Bocci, V.; Bondar, A.; Bondar, N.; Bonivento, W.; Borghi, S.; Borgia, A.; Borsato, M.; Bowcock, T. J. V.; Bowen, E.; Bozzi, C.; Brett, D.; Britsch, M.; Britton, T.; Brodzicka, J.; Brook, N. H.; Bursche, A.; Buytaert, J.; Cadeddu, S.; Calabrese, R.; Calvi, M.; Calvo Gomez, M.; Campana, P.; Campora Perez, D.; Capriotti, L.; Carbone, A.; Carboni, G.; Cardinale, R.; Cardini, A.; Carniti, P.; Carson, L.; Carvalho Akiba, K.; Casanova Mohr, R.; Casse, G.; Cassina, L.; Castillo Garcia, L.; Cattaneo, M.; Cauet, Ch.; Cavallero, G.; Cenci, R.; Charles, M.; Charpentier, Ph.; Chefdeville, M.; Chen, S.; Cheung, S.-F.; Chiapolini, N.; Chrzaszcz, M.; Cid Vidal, X.; Ciezarek, G.; Clarke, P. E. L.; Clemencic, M.; Cliff, H. V.; Closier, J.; Coco, V.; Cogan, J.; Cogneras, E.; Cogoni, V.; Cojocariu, L.; Collazuol, G.; Collins, P.; Comerma-Montells, A.; Contu, A.; Cook, A.; Coombes, M.; Coquereau, S.; Corti, G.; Corvo, M.; Counts, I.; Couturier, B.; Cowan, G. A.; Craik, D. C.; Crocombe, A. C.; Cruz Torres, M.; Cunliffe, S.; Currie, R.; D'Ambrosio, C.; Dalseno, J.; David, P.; David, P. N. Y.; Davis, A.; De Bruyn, K.; De Capua, S.; De Cian, M.; De Miranda, J. M.; De Paula, L.; De Silva, W.; De Simone, P.; Dean, C.-T.; Decamp, D.; Deckenhoff, M.; Del Buono, L.; Déléage, N.; Derkach, D.; Deschamps, O.; Dettori, F.; Dey, B.; Di Canto, A.; Di Ruscio, F.; Dijkstra, H.; Donleavy, S.; Dordei, F.; Dorigo, M.; Dosil Suárez, A.; Dossett, D.; Dovbnya, A.; Dreimanis, K.; Dujany, G.; Dupertuis, F.; Durante, P.; Dzhelyadin, R.; Dziurda, A.; Dzyuba, A.; Easo, S.; Egede, U.; Egorychev, V.; Eidelman, S.; Eisenhardt, S.; Eitschberger, U.; Ekelhof, R.; Eklund, L.; El Rifai, I.; Elsasser, Ch.; Ely, S.; Esen, S.; Evans, H. M.; Evans, T.; Falabella, A.; Färber, C.; Farinelli, C.; Farley, N.; Farry, S.; Fay, R.; Ferguson, D.; Fernandez Albor, V.; Ferreira Rodrigues, F.; Ferro-Luzzi, M.; Filippov, S.; Fiore, M.; Fiorini, M.; Firlej, M.; Fitzpatrick, C.; Fiutowski, T.; Fol, P.; Fontana, M.; Fontanelli, F.; Forty, R.; Francisco, O.; Frank, M.; Frei, C.; Frosini, M.; Fu, J.; Furfaro, E.; Gallas Torreira, A.; Galli, D.; Gallorini, S.; Gambetta, S.; Gandelman, M.; Gandini, P.; Gao, Y.; García Pardiñas, J.; Garofoli, J.; Garra Tico, J.; Garrido, L.; Gascon, D.; Gaspar, C.; Gastaldi, U.; Gauld, R.; Gavardi, L.; Gazzoni, G.; Geraci, A.; Gersabeck, E.; Gersabeck, M.; Gershon, T.; Ghez, Ph.; Gianelle, A.; Gianì, S.; Gibson, V.; Giubega, L.; Gligorov, V. V.; Göbel, C.; Golubkov, D.; Golutvin, A.; Gomes, A.; Gotti, C.; Grabalosa Gándara, M.; Graciani Diaz, R.; Granado Cardoso, L. A.; Graugés, E.; Graverini, E.; Graziani, G.; Grecu, A.; Greening, E.; Gregson, S.; Griffith, P.; Grillo, L.; Grünberg, O.; Gui, B.; Gushchin, E.; Guz, Yu.; Gys, T.; Hadjivasiliou, C.; Haefeli, G.; Haen, C.; Haines, S. C.; Hall, S.; Hamilton, B.; Hampson, T.; Han, X.; Hansmann-Menzemer, S.; Harnew, N.; Harnew, S. T.; Harrison, J.; He, J.; Head, T.; Heijne, V.; Hennessy, K.; Henrard, P.; Henry, L.; Hernando Morata, J. A.; van Herwijnen, E.; Heß, M.; Hicheur, A.; Hill, D.; Hoballah, M.; Hombach, C.; Hulsbergen, W.; Humair, T.; Hussain, N.; Hutchcroft, D.; Hynds, D.; Idzik, M.; Ilten, P.; Jacobsson, R.; Jaeger, A.; Jalocha, J.; Jans, E.; Jawahery, A.; Jing, F.; John, M.; Johnson, D.; Jones, C. R.; Joram, C.; Jost, B.; Jurik, N.; Kandybei, S.; Kanso, W.; Karacson, M.; Karbach, T. M.; Karodia, S.; Kelsey, M.; Kenyon, I. R.; Kenzie, M.; Ketel, T.; Khanji, B.; Khurewathanakul, C.; Klaver, S.; Klimaszewski, K.; Kochebina, O.; Kolpin, M.; Komarov, I.; Koopman, R. F.; Koppenburg, P.; Korolev, M.; Kravchuk, L.; Kreplin, K.; Kreps, M.; Krocker, G.; Krokovny, P.; Kruse, F.; Kucewicz, W.; Kucharczyk, M.; Kudryavtsev, V.; Kurek, K.; Kvaratskheliya, T.; La Thi, V. N.; Lacarrere, D.; Lafferty, G.; Lai, A.; Lambert, D.; Lambert, R. W.; Lanfranchi, G.; Langenbruch, C.; Langhans, B.; Latham, T.; Lazzeroni, C.; Le Gac, R.; van Leerdam, J.; Lees, J.-P.; Lefèvre, R.; Leflat, A.; Lefrançois, J.; Leroy, O.; Lesiak, T.; Leverington, B.; Li, Y.; Likhomanenko, T.; Liles, M.; Lindner, R.; Linn, C.; Lionetto, F.; Liu, B.; Lohn, S.; Longstaff, I.; Lopes, J. H.; Lowdon, P.; Lucchesi, D.; Luo, H.; Lupato, A.; Luppi, E.; Lupton, O.; Machefert, F.; Machikhiliyan, I. V.; Maciuc, F.; Maev, O.; Malde, S.; Malinin, A.; Manca, G.; Mancinelli, G.; Manning, P.; Mapelli, A.; Maratas, J.; Marchand, J. F.; Marconi, U.; Marin Benito, C.; Marino, P.; Märki, R.; Marks, J.; Martellotti, G.; Martinelli, M.; Martinez Santos, D.; Martinez Vidal, F.; Martins Tostes, D.; Massafferri, A.; Matev, R.; Mathe, Z.; Matteuzzi, C.; Mauri, A.; Maurin, B.; Mazurov, A.; McCann, M.; McCarthy, J.; McNab, A.; McNulty, R.; McSkelly, B.; Meadows, B.; Meier, F.; Meissner, M.; Merk, M.; Milanes, D. A.; Minard, M.-N.; Molina Rodriguez, J.; Monteil, S.; Morandin, M.; Morawski, P.; Mordà, A.; Morello, M. J.; Moron, J.; Morris, A.-B.; Mountain, R.; Muheim, F.; Müller, K.; Mussini, M.; Muster, B.; Naik, P.; Nakada, T.; Nandakumar, R.; Nasteva, I.; Needham, M.; Neri, N.; Neubert, S.; Neufeld, N.; Neuner, M.; Nguyen, A. D.; Nguyen, T. D.; Nguyen-Mau, C.; Nicol, M.; Niess, V.; Niet, R.; Nikitin, N.; Nikodem, T.; Novoselov, A.; O'Hanlon, D. P.; Oblakowska-Mucha, A.; Obraztsov, V.; Ogilvy, S.; Okhrimenko, O.; Oldeman, R.; Onderwater, C. J. G.; Osorio Rodrigues, B.; Otalora Goicochea, J. M.; Otto, A.; Owen, P.; Oyanguren, A.; Pal, B. K.; Palano, A.; Palombo, F.; Palutan, M.; Panman, J.; Papanestis, A.; Pappagallo, M.; Pappalardo, L. L.; Parkes, C.; Parkinson, C. J.; Passaleva, G.; Patel, G. D.; Patel, M.; Patrignani, C.; Pearce, A.; Pellegrino, A.; Penso, G.; Pepe Altarelli, M.; Perazzini, S.; Perret, P.; Pescatore, L.; Petridis, K.; Petrolini, A.; Picatoste Olloqui, E.; Pietrzyk, B.; Pilař, T.; Pinci, D.; Pistone, A.; Playfer, S.; Plo Casasus, M.; Polci, F.; Poluektov, A.; Polyakov, I.; Polycarpo, E.; Popov, A.; Popov, D.; Popovici, B.; Potterat, C.; Price, E.; Price, J. D.; Prisciandaro, J.; Pritchard, A.; Prouve, C.; Pugatch, V.; Puig Navarro, A.; Punzi, G.; Qian, W.; Quagliani, R.; Rachwal, B.; Rademacker, J. H.; Rakotomiaramanana, B.; Rama, M.; Rangel, M. S.; Raniuk, I.; Rauschmayr, N.; Raven, G.; Redi, F.; Reichert, S.; Reid, M. M.; dos Reis, A. C.; Ricciardi, S.; Richards, S.; Rihl, M.; Rinnert, K.; Rives Molina, V.; Robbe, P.; Rodrigues, A. B.; Rodrigues, E.; Rodriguez Perez, P.; Roiser, S.; Romanovsky, V.; Romero Vidal, A.; Rotondo, M.; Rouvinet, J.; Ruf, T.; Ruiz, H.; Ruiz Valls, P.; Saborido Silva, J. J.; Sagidova, N.; Sail, P.; Saitta, B.; Salustino Guimaraes, V.; Sanchez Mayordomo, C.; Sanmartin Sedes, B.; Santacesaria, R.; Santamarina Rios, C.; Santovetti, E.; Sarti, A.; Satriano, C.; Satta, A.; Saunders, D. M.; Savrina, D.; Schiller, M.; Schindler, H.; Schlupp, M.; Schmelling, M.; Schmidt, B.; Schneider, O.; Schopper, A.; Schune, M.-H.; Schwemmer, R.; Sciascia, B.; Sciubba, A.; Semennikov, A.; Sepp, I.; Serra, N.; Serrano, J.; Sestini, L.; Seyfert, P.; Shapkin, M.; Shapoval, I.; Shcheglov, Y.; Shears, T.; Shekhtman, L.; Shevchenko, V.; Shires, A.; Silva Coutinho, R.; Simi, G.; Sirendi, M.; Skidmore, N.; Skillicorn, I.; Skwarnicki, T.; Smith, N. A.; Smith, E.; Smith, E.; Smith, J.; Smith, M.; Snoek, H.; Sokoloff, M. D.; Soler, F. J. P.; Soomro, F.; Souza, D.; Souza De Paula, B.; Spaan, B.; Spradlin, P.; Sridharan, S.; Stagni, F.; Stahl, M.; Stahl, S.; Steinkamp, O.; Stenyakin, O.; Sterpka, F.; Stevenson, S.; Stoica, S.; Stone, S.; Storaci, B.; Stracka, S.; Straticiuc, M.; Straumann, U.; Stroili, R.; Sun, L.; Sutcliffe, W.; Swientek, K.; Swientek, S.; Syropoulos, V.; Szczekowski, M.; Szczypka, P.; Szumlak, T.; T'Jampens, S.; Teklishyn, M.; Tellarini, G.; Teubert, F.; Thomas, C.; Thomas, E.; van Tilburg, J.; Tisserand, V.; Tobin, M.; Todd, J.; Tolk, S.; Tomassetti, L.; Tonelli, D.; Topp-Joergensen, S.; Torr, N.; Tournefier, E.; Tourneur, S.; Trabelsi, K.; Tran, M. T.; Tresch, M.; Trisovic, A.; Tsaregorodtsev, A.; Tsopelas, P.; Tuning, N.; Ubeda Garcia, M.; Ukleja, A.; Ustyuzhanin, A.; Uwer, U.; Vacca, C.; Vagnoni, V.; Valenti, G.; Vallier, A.; Vazquez Gomez, R.; Vazquez Regueiro, P.; Vázquez Sierra, C.; Vecchi, S.; Velthuis, J. J.; Veltri, M.; Veneziano, G.; Vesterinen, M.; Viana Barbosa, J. V.; Viaud, B.; Vieira, D.; Vieites Diaz, M.; Vilasis-Cardona, X.; Vollhardt, A.; Volyanskyy, D.; Voong, D.; Vorobyev, A.; Vorobyev, V.; Voß, C.; de Vries, J. A.; Waldi, R.; Wallace, C.; Wallace, R.; Walsh, J.; Wandernoth, S.; Wang, J.; Ward, D. R.; Watson, N. K.; Websdale, D.; Whitehead, M.; Wiedner, D.; Wilkinson, G.; Wilkinson, M.; Williams, M. P.; Williams, M.; Wilschut, H. W.; Wilson, F. F.; Wimberley, J.; Wishahi, J.; Wislicki, W.; Witek, M.; Wormser, G.; Wotton, S. A.; Wright, S.; Wyllie, K.; Xie, Y.; Xing, Z.; Xu, Z.; Yang, Z.; Yuan, X.; Yushchenko, O.; Zangoli, M.; Zavertyaev, M.; Zhang, L.; Zhang, W. C.; Zhang, Y.; Zhelezov, A.; Zhokhov, A.; Zhong, L.

    2015-07-01

    An angular analysis of the decay is performed using pp collisions corresponding to an integrated luminosity of 1 .0 fb-1 collected by the LHCb experiment at a centre-of-mass energy TeV. A combined angular and mass analysis separates six helicity amplitudes and allows the measurement of the longitudinal polarisation fraction f L = 0 .201 ± 0 .057 (stat .) ± 0 .040 (syst .) for the decay. A large scalar contribution from the K {0/∗}(1430) and K {0/∗}(800) resonances is found, allowing the determination of additional CP asymmetries. Triple product and direct CP asymmetries are determined to be compatible with the Standard Model expectations. The branching fraction is measured to be (10 .8 ± 2 .1 (stat .) ± 1 .4 (syst .) ± 0 .6 ( f d /f s )) × 10-6. [Figure not available: see fulltext.

  19. Denoising time-domain induced polarisation data using wavelet techniques

    NASA Astrophysics Data System (ADS)

    Deo, Ravin N.; Cull, James P.

    2016-05-01

    Time-domain induced polarisation (TDIP) methods are routinely used for near-surface evaluations in quasi-urban environments harbouring networks of buried civil infrastructure. A conventional technique for improving signal to noise ratio in such environments is by using analogue or digital low-pass filtering followed by stacking and rectification. However, this induces large distortions in the processed data. In this study, we have conducted the first application of wavelet based denoising techniques for processing raw TDIP data. Our investigation included laboratory and field measurements to better understand the advantages and limitations of this technique. It was found that distortions arising from conventional filtering can be significantly avoided with the use of wavelet based denoising techniques. With recent advances in full-waveform acquisition and analysis, incorporation of wavelet denoising techniques can further enhance surveying capabilities. In this work, we present the rationale for utilising wavelet denoising methods and discuss some important implications, which can positively influence TDIP methods.

  20. The more we know, the more we have to discover: an exciting future for understanding cilia and ciliopathies.

    PubMed

    Benmerah, Alexandre; Durand, Bénédicte; Giles, Rachel H; Harris, Tess; Kohl, Linda; Laclef, Christine; Meilhac, Sigolène M; Mitchison, Hannah M; Pedersen, Lotte B; Roepman, Ronald; Swoboda, Peter; Ueffing, Marius; Bastin, Philippe

    2015-01-01

    The Cilia 2014 conference was organised by four European networks: the Ciliopathy Alliance, the Groupement de Recherche CIL, the Nordic Cilia and Centrosome Network and the EU FP7 programme SYSCILIA. More than 400 delegates from 27 countries gathered at the Institut Pasteur conference centre in Paris, including 30 patients and patient representatives. The meeting offered a unique opportunity for exchange between different scientific and medical communities. Major highlights included new discoveries about the roles of motile and immotile cilia during development and homeostasis, the mechanism of cilium construction, as well as progress in diagnosis and possible treatment of ciliopathies. The contributions to the cilia field of flagellated infectious eukaryotes and of systems biology were also presented. PMID:25974046

  1. Axial super-localisation using rotating point spread functions shaped by polarisation-dependent phase modulation.

    PubMed

    Roider, Clemens; Jesacher, Alexander; Bernet, Stefan; Ritsch-Marte, Monika

    2014-02-24

    We present an approach for point spread function (PSF) engineering that allows one to shape the optical wavefront independently in both polarisation directions, with two adjacent phase masks displayed on a single liquid-crystal spatial light modulator (LC-SLM). The set-up employs a polarising beam splitter and a geometric image rotator to rectify and process both polarisation directions detected by the camera. We shape a single-lobe ("corkscrew") PSF that rotates upon defocus for each polarisation channel and combine the two polarisation channels with a relative 180° phase-shift on the computer, merging them into a single PSF that exhibits two lobes whose orientation contains information about the axial position. A major advantage lies in the possibility to measure and eliminate the aberrations in the two polarisation channels independently. We demonstrate axial super-localisation of isotropically emitting fluorescent nanoparticles. Our implementation of the single-lobe PSFs follows the method proposed by Prasad [Opt. Lett.38, 585 (2013)], and thus is to the best of our knowledge the first experimental realisation of this suggestion. For comparison we also study an approach with a rotating double-helix PSFs (in only one polarisation channel) and ascertain the trade-off between localisation precision and axial working range. PMID:24663724

  2. A continuous roll-pulling approach for the fabrication of magnetic artificial cilia with microfluidic pumping capability.

    PubMed

    Wang, Ye; den Toonder, Jaap; Cardinaels, Ruth; Anderson, Patrick

    2016-06-21

    Magnetic artificial cilia are micro-hairs covering a surface that can be actuated using a time-dependent magnetic field to pump or mix fluids in microfluidic devices. This paper presents a novel fabrication method to realize magnetic artificial cilia using a roll-pulling process, in which a cylinder decorated with micro-pillars rolls over a liquid precursor film that contains magnetic particles at a speed up to 1 m s(-1), while a magnetic field is applied. Due to the interaction between the pillars and the liquid film, micro-hairs are pulled out of the film. In this way, surfaces with slender cone-shaped magnetic artificial cilia were produced. When integrated in a closed-loop channel, the artificial cilia were shown to be capable of generating substantial microfluidic pumping using external magnetic actuation. The spatial arrangement of the cilia can be varied by altering the layout of the micro-pillars on the roll surface. In addition, the final geometry of the individual cilia depends on the rheological properties of the precursor material in combination with the processing parameters of the roll-pulling process. A rheological study and fabrication tests were carried out for a range of precursor material compositions to obtain insight into the relation between precursor rheology and processing conditions on the one hand, and cilia geometry on the other hand. The development of this cleanroom-free, high speed and potentially large area method of production of artificial cilia is another step towards their implementation in real-life applications. PMID:27210071

  3. The viewing angle effect on Hα-line impact polarisation in impulsive solar events

    NASA Astrophysics Data System (ADS)

    Zharkova, V. V.; Kashapova, L. K.

    2005-03-01

    The effect of a viewing angle on the hydroden Hα-line impact polarisation is investigated in a plane vertical atmosphere arbitrary located on the solar disk. The impact polarisation is assumed to be caused by precipitating beam electrons with pitch-angular anisotropy steadily injected into the flaring atmosphere from its top. The polarisation is calculated for a 3 level plus continuum hydrogen atom affected by Zeeman splitting in a moderate magnetic field taking into account depolarising effects of diffusive radiation and collisions with thermal electrons. The Hα polarisation profiles are affected by electron beams only in the line cores whereas the wings are fully depolarized by the collisions with thermal electrons despite the extended wing emission, or “moustaches”, caused by beam electrons. The full (integrated in wavelength) Hα-line linear polarisation, caused by moderate electron beams, is shown to be 2{-}20% and either negative or positive depending on the position of a flaring loop on the solar disk and the direction of an emitted photon from the local magnetic field. The polarisation plane is projected onto a viewing angle psi, being a superposition of the flare location on a solar disk and the magnetic field deviation from vertical on the solar surface. For viewing angles less then 50^circ the Hα-line impact polarisation is negative increasing up to -10% towards smaller angles, meaning that the polarisation is mostly perpendicular to the plane {B}× {K} where B is the magnetic field induction and K is the photon momentum vector. For viewing angles greater than 60^circ the measured impact polarisation becomes positive, sharply increasing up to 20% towards the limb. In the range of 50{-}60^circ the observed impact polarisation goes through a zero point despite the actual presence of beam electrons in the flaring atmosphere. The theoretical predictions of the dependence of polarisation degree on viewing angle fit remarkably well the observations of

  4. Polarisation effects in twin-core fibre: Application for mode locking in a fibre laser

    SciTech Connect

    Lobach, I A; Kablukov, S I; Podivilov, Evgenii V; Babin, Sergei A; Apolonski, A A

    2012-09-30

    We report the first measurements of the longitudinal power distribution in a twin-core optical fibre at different input light polarisations. Experimental evidence is presented that, because of the difference in birefringence between the cores, the power in them depends on which core the beam is launched into. Experimental data are interpreted in terms of a modified polarisation model for mode coupling in twin-core fibres which takes into account the birefringence of the cores. In addition, we demonstrate for the first time the use of the polarisation properties of a twincore fibre for mode locking in a fibre laser. (optical fibres, lasers and amplifiers. properties and applications)

  5. Birefringence in anisotropic optical fibres studied by polarised light Brillouin reflectometry

    SciTech Connect

    Smirnov, A S; Burdin, V V; Konstantinov, Yu A; Petukhov, A S; Drozdov, I R; Kuz'minykh, Ya S; Besprozvannykh, V G

    2015-01-31

    Modal birefringence (the difference between the effective refractive indices of orthogonal polarisation modes) is one of the key parameters of anisotropic single-mode fibres, characterising their ability to preserve a linearly polarised state of input light. This parameter is commonly measured using short pieces of fibre, but such procedures are destructive and allow the birefringence to be determined only at the ends of long fibres. In this study, polarised light Brillouin reflectometry is used to assess birefringence uniformity throughout the length of an anisotropic fibre. (optical fibres)

  6. Capillary compressor of femtosecond laser pulses with nonlinear rotation of polarisation ellipse

    SciTech Connect

    Konyashchenko, Aleksandr V; Kostryukov, P V; Losev, Leonid L; Tenyakov, S Yu

    2012-03-31

    The process of nonlinear rotation of the polarisation ellipse of laser radiation, occurring simultaneously with the broadening of the pulse spectrum due to nonlinear self-phase modulation in a gas-filled capillary, is studied. It is shown that the maximal rotation of the polarisation ellipse is experienced by the spectral components, shifted towards the short-wavelength side with respect to the central wavelength of the initial laser pulse. Using the effect of polarisation ellipse rotation, an eightfold increase in the energy contrast ratio of a 28-fs light pulse, obtained by compression of the radiation pulse from an ytterbium laser with the duration 290 fs, is implemented.

  7. Changes of lymphocyte membrane fluidity in rheumatoid arthritis: a fluorescence polarisation study.

    PubMed Central

    Beccerica, E; Piergiacomi, G; Curatola, G; Ferretti, G

    1988-01-01

    Fluorescence polarisation of 1,6-diphenyl-1,3,5-hexatriene was used to study the lymphocyte membrane in rheumatoid arthritis. The increase of polarisation value in the patients (n = 27) compared with healthy controls (n = 32) suggests a decrease of membrane fluidity. Moreover, erythrocyte sedimentation rate (ESR) and plasma fibrinogen concentrations were positively correlated with lymphocyte fluorescence polarisation values (r = 0.66 and r = 0.76 respectively). The results suggest that the changes in lymphocyte membrane fluidity could be involved in the pathogenetic mechanism of rheumatoid arthritis. PMID:3382266

  8. Polarisation and compression of {sup 3}He for Magnetic Resonance Imaging purposes

    SciTech Connect

    Geurts, D. G.; Brand, J. F. J. van den; Bulten, H. J.; Poolman, H. R.; Ferro-Luzzi, M.; Nicolay, K.

    1998-01-20

    Magnetic Resonance Imaging is often used in medical science as a diagnostic tool for the human body. Conventional MRI uses the NMR signal from the protons of water molecules in tissue to image the interior of the patient's body. However, for certain areas such as the lungs and airways, the usage of a highly polarised gas yields better results. We are currently constructing an apparatus that uses polarised {sup 3}He gas to produce detailed images of those signal-deficient moyeties. We also plan to study possible uptake of polarised {sup 3}He gas by the circulatory system to image other organs.

  9. Collective motion of motile cilia: from human airways to model systems

    NASA Astrophysics Data System (ADS)

    Cicuta, Pietro; Feriani, Luigi; Chioccioli, Maurizio; Kotar, Jurij

    Mammalian airways are a fantastic playground of nonlinear phenomena, from the function of individual active filaments, to the emerging collective behaviour, to the rheology of the mucus solution surrounding cilia. We have been investigating the fundamental physics of this system through a variety of model system approaches, both experimental and computational. In the last year we have started measurements on living human cells, observing cilia shape during beating, and measuring speed and coherence of the collective dynamics. We report on significant differences in the collective motion in ciliated cell carpets from a variety of diseases, and we attempt to reconcile the collective dynamical phenotypes to the properties of individual filaments and the mechanics of the environment.

  10. Ca2+-dependent regulation of beat frequency of cilia in Paramecium.

    PubMed

    Nakaoka, Y; Tanaka, H; Oosawa, F

    1984-01-01

    Triton-extracted models of Paramecium cells prepared in the presence of Mg2+ and EGTA showed Ca2+ sensitivity not only in the direction of beat but also in the beat frequency of cilia. The beat frequency increased over a range of concentration of Ca2+ from 2 X 10(-7) to 4 X 10(-7) M, in which the model swam forwards. The frequency increased also above 10(-6)M-Ca2+, in which the model swam backwards. The increase in frequency was inhibited by calmodulin antagonists. Intracellular injection of a Ca2+ buffer giving a free Ca2+ concentration of 2 X 10(-7) to 5 X 10(-7) M in intact cells induced an increase in the beat frequency. Therefore, it is very likely that the beat frequency of cilia is regulated by the intracellular concentration of Ca2+. PMID:6715425

  11. Katanin p80 Regulates Human Cortical Development by Limiting Centriole and Cilia Number

    PubMed Central

    Hu, Wen F.; Pomp, Oz; Ben-Omran, Tawfeg; Kodani, Andrew; Henke, Katrin; Mochida, Ganeshwaran H.; Yu, Timothy W.; Woodworth, Mollie B.; Bonnard, Carine; Raj, Grace Selva; Tan, Thong Teck; Hamamy, Hanan; Masri, Amira; Shboul, Mohammad; Al Saffar, Muna; Partlow, Jennifer N.; Al-Dosari, Mohammed; Alazami, Anas; Alowain, Mohammed; Alkuraya, Fowzan S.; Reiter, Jeremy F.; Harris, Matthew P.; Reversade, Bruno; Walsh, Christopher A.

    2015-01-01

    SUMMARY Katanin is a microtubule-severing complex whose catalytic activities are well characterized, but whose in vivo functions are incompletely understood. Human mutations in KATNB1, which encodes the noncatalytic regulatory p80 subunit of katanin, cause severe microlissencephaly. Loss of Katnb1 in mice confirms essential roles in neurogenesis and cell survival, while loss of zebrafish katnb1 reveals specific roles for katnin p80 in early and late developmental stages. Surprisingly, Katnb1 null mutant mouse embryos display hallmarks of aberrant Sonic hedgehog signaling, including holoprosencephaly. KATNB1-deficient human cells show defective proliferation and spindle structure, while Katnb1 null fibroblasts also demonstrate a remarkable excess of centrioles, with supernumerary cilia but deficient Hedgehog signaling. Our results reveal unexpected functions for KATNB1 in regulating overall centriole, mother centriole, and cilia number, and as an essential gene for normal Hedgehog signaling during neocortical development. PMID:25521379

  12. The sense of smell, its signalling pathways, and the dichotomy of cilia and microvilli in olfactory sensory cells

    PubMed Central

    2007-01-01

    Smell is often regarded as an ancillary perception in primates, who seem so dominated by their sense of vision. In this paper, we will portray some aspects of the significance of olfaction to human life and speculate on what evolutionary factors contribute to keeping it alive. We then outline the functional architecture of olfactory sensory neurons and their signal transduction pathways, which are the primary detectors that render olfactory perception possible. Throughout the phylogenetic tree, olfactory neurons, at their apical tip, are either decorated with cilia or with microvilli. The significance of this dichotomy is unknown. It is generally assumed that mammalian olfactory neurons are of the ciliary type only. The existance of so-called olfactory microvillar cells in mammals, however, is well documented, but their nature remains unclear and their function orphaned. This paper discusses the possibility, that in the main olfactory epithelium of mammals ciliated and microvillar sensory cells exist concurrently. We review evidence related to this hypothesis and ask, what function olfactory microvillar cells might have and what signalling mechanisms they use. PMID:17903277

  13. Hsp40 is involved in cilia regeneration in sea urchin embryos.

    PubMed

    Casano, Caterina; Gianguzza, Fabrizio; Roccheri, Maria C; Di Giorgi, Rossana; Maenza, Luigia; Ragusa, Maria A

    2003-12-01

    In a previous paper we demonstrated that, in Paracentrotus lividus embryos, deciliation represents a specific kind of stress that induces an increase in the levels of an acidic protein of about 40 kD (p40). Here we report that deciliation also induces an increase in Hsp40 chaperone levels and enhancement of its ectodermal localization. We suggest that Hsp40 might play a chaperoning role in cilia regeneration. PMID:14623926

  14. Global genetic analysis in mice unveils central role for cilia in congenital heart disease.

    PubMed

    Li, You; Klena, Nikolai T; Gabriel, George C; Liu, Xiaoqin; Kim, Andrew J; Lemke, Kristi; Chen, Yu; Chatterjee, Bishwanath; Devine, William; Damerla, Rama Rao; Chang, Chienfu; Yagi, Hisato; San Agustin, Jovenal T; Thahir, Mohamed; Anderton, Shane; Lawhead, Caroline; Vescovi, Anita; Pratt, Herbert; Morgan, Judy; Haynes, Leslie; Smith, Cynthia L; Eppig, Janan T; Reinholdt, Laura; Francis, Richard; Leatherbury, Linda; Ganapathiraju, Madhavi K; Tobita, Kimimasa; Pazour, Gregory J; Lo, Cecilia W

    2015-05-28

    Congenital heart disease (CHD) is the most prevalent birth defect, affecting nearly 1% of live births; the incidence of CHD is up to tenfold higher in human fetuses. A genetic contribution is strongly suggested by the association of CHD with chromosome abnormalities and high recurrence risk. Here we report findings from a recessive forward genetic screen in fetal mice, showing that cilia and cilia-transduced cell signalling have important roles in the pathogenesis of CHD. The cilium is an evolutionarily conserved organelle projecting from the cell surface with essential roles in diverse cellular processes. Using echocardiography, we ultrasound scanned 87,355 chemically mutagenized C57BL/6J fetal mice and recovered 218 CHD mouse models. Whole-exome sequencing identified 91 recessive CHD mutations in 61 genes. This included 34 cilia-related genes, 16 genes involved in cilia-transduced cell signalling, and 10 genes regulating vesicular trafficking, a pathway important for ciliogenesis and cell signalling. Surprisingly, many CHD genes encoded interacting proteins, suggesting that an interactome protein network may provide a larger genomic context for CHD pathogenesis. These findings provide novel insights into the potential Mendelian genetic contribution to CHD in the fetal population, a segment of the human population not well studied. We note that the pathways identified show overlap with CHD candidate genes recovered in CHD patients, suggesting that they may have relevance to the more complex genetics of CHD overall. These CHD mouse models and >8,000 incidental mutations have been sperm archived, creating a rich public resource for human disease modelling. PMID:25807483

  15. WD60/FAP163 is a dynein intermediate chain required for retrograde intraflagellar transport in cilia.

    PubMed

    Patel-King, Ramila S; Gilberti, Renée M; Hom, Erik F Y; King, Stephen M

    2013-09-01

    Retrograde intraflagellar transport (IFT) is required for assembly of cilia. We identify a Chlamydomonas flagellar protein (flagellar-associated protein 163 [FAP163]) as being closely related to the D1bIC(FAP133) intermediate chain (IC) of the dynein that powers this movement. Biochemical analysis revealed that FAP163 is present in the flagellar matrix and is actively trafficked by IFT. Furthermore, FAP163 copurified with D1bIC(FAP133) and the LC8 dynein light chain, indicating that it is an integral component of the retrograde IFT dynein. To assess the functional role of FAP163, we generated an RNA interference knockdown of the orthologous protein (WD60) in planaria. The Smed-wd60(RNAi) animals had a severe ciliary assembly defect that dramatically compromised whole-organism motility. Most cilia were present as short stubs that had accumulated large quantities of IFT particle-like material between the doublet microtubules and the membrane. The few remaining approximately full-length cilia had a chaotic beat with a frequency reduced from 24 to ∼10 Hz. Thus WD60/FAP163 is a dynein IC that is absolutely required for retrograde IFT and ciliary assembly. PMID:23864713

  16. A Computational Model of Dynein Activation Patterns that Can Explain Nodal Cilia Rotation

    PubMed Central

    Chen, Duanduan; Zhong, Yi

    2015-01-01

    Normal left-right patterning in vertebrates depends on the rotational movement of nodal cilia. In order to produce this ciliary motion, the activity of axonemal dyneins must be tightly regulated in a temporal and spatial manner; the specific activation pattern of the dynein motors in the nodal cilia has not been reported. Contemporary imaging techniques cannot directly assess dynein activity in a living cilium. In this study, we establish a three-dimensional model to mimic the ciliary ultrastructure and assume that the activation of dynein proteins is related to the interdoublet distance. By employing finite-element analysis and grid deformation techniques, we simulate the mechanical function of dyneins by pairs of point loads, investigate the time-variant interdoublet distance, and simulate the dynein-triggered ciliary motion. The computational results indicate that, to produce the rotational movement of nodal cilia, the dynein activity is transferred clockwise (looking from the tip) between the nine doublet microtubules, and along each microtubule, the dynein activation should occur faster at the basal region and slower when it is close to the ciliary tip. Moreover, the time cost by all the dyneins along one microtubule to be activated can be used to deduce the dynein activation pattern; it implies that, as an alternative method, measuring this time can indirectly reveal the dynein activity. The proposed protein-structure model can simulate the ciliary motion triggered by various dynein activation patterns explicitly and may contribute to furthering the studies on axonemal dynein activity. PMID:26153700

  17. A complex of BBS1 and NPHP7 is required for cilia motility in zebrafish.

    PubMed

    Kim, Yun Hee; Epting, Daniel; Slanchev, Krasimir; Engel, Christina; Walz, Gerd; Kramer-Zucker, Albrecht

    2013-01-01

    Bardet-Biedl syndrome (BBS) and nephronophthisis (NPH) are hereditary autosomal recessive disorders, encoded by two families of diverse genes. BBS and NPH display several overlapping phenotypes including cystic kidney disease, retinitis pigmentosa, liver fibrosis, situs inversus and cerebellar defects. Since most of the BBS and NPH proteins localize to cilia and/or their appendages, BBS and NPH are considered ciliopathies. In this study, we characterized the function of the transcription factor Nphp7 in zebrafish, and addressed the molecular connection between BBS and NPH. The knockdown of zebrafish bbs1 and nphp7.2 caused similar phenotypic changes including convergent extension defects, curvature of the body axis, hydrocephalus, abnormal heart looping and cystic pronephros, all consistent with an altered ciliary function. Immunoprecipitation assays revealed a physical interaction between BBS1 and NPHP7, and the simultaneous knockdown of zbbs1 and znphp7.2 enhanced the cystic pronephros phenotype synergistically, suggesting a genetic interaction between zbbs1 and znphp7.2 in vivo. Deletion of zBbs1 or zNphp7.2 did not compromise cilia formation, but disrupted cilia motility. Although NPHP7 has been shown to act as transcriptional repressor, our studies suggest a crosstalk between BBS1 and NPHP7 in regulating normal function of the cilium. PMID:24069149

  18. Congenital Heart Disease Genetics Uncovers Context-Dependent Organization and Function of Nucleoporins at Cilia.

    PubMed

    Del Viso, Florencia; Huang, Fang; Myers, Jordan; Chalfant, Madeleine; Zhang, Yongdeng; Reza, Nooreen; Bewersdorf, Joerg; Lusk, C Patrick; Khokha, Mustafa K

    2016-09-12

    Human genomics is identifying candidate genes for congenital heart disease (CHD), but discovering the underlying mechanisms remains challenging. In a patient with CHD and heterotaxy (Htx), a disorder of left-right patterning, we previously identified a duplication in Nup188. However, a mechanism to explain how a component of the nuclear pore complex (NPC) could cause Htx/CHD was undefined. Here, we show that knockdown of Nup188 or its binding partner Nup93 leads to a loss of cilia during embryonic development while leaving NPC function largely intact. Many data, including the localization of endogenous Nup188/93 at cilia bases, support their direct role at cilia. Super-resolution imaging of Nup188 shows two barrel-like structures with dimensions and organization incompatible with an NPC-like ring, arguing against a proposed "ciliary pore complex." We suggest that the nanoscale organization and function of nucleoporins are context dependent in a way that is required for the structure of the heart. PMID:27593162

  19. The intraflagellar transport protein IFT80 is required for cilia formation and osteogenesis

    PubMed Central

    Yang, Shuying; Wang, Changdong

    2012-01-01

    Intraflagellar transport (IFT) proteins are essential for the assembly and maintenance of cilia, which play important roles in development and homeostasis. IFT80 is a newly defined IFT protein. Partial mutation of IFT80 in humans causes diseases such as Jeune asphyxiating thoracic dystrophy (JATD) and short rib polydactyly (SRP) type III with abnormal skeletal development. However, the role and mechanism of IFT80 in osteogenesis is unknown. Here, we first detected IFT80 expression pattern and found that IFT80 was highly expressed in mouse long bone, skull, and during osteoblast differentiation. By using lentivirus-mediated RNA interference (RNAi) technology to silence IFT80 in murine mesenchymal progenitor cell line-C3H10T1/2 and bone marrow derived stromal cells, we found that silencing IFT80 led to either shortening or loss of cilia and the decrease of Arl13b expression - a small GTPase that is localized in cilia. Additionally, silencing IFT80 blocked the expression of osteoblast markers and significantly inhibited ALP activity and cell mineralization. We further found that IFT80 silencing inhibited the expression of Gli2, a critical transcriptional factor in the hedgehog signaling pathway. Overexpression of Gli2 rescued the deficiency of osteoblast differentiation from IFT80-silenced cells, and dramatically promoted osteoblast differentiation. Moreover, introduction of Smo agonist (SAG) promotes osteoblast differentiation, which was partially inhibited by IFT80 silencing. Thus, these results suggested that IFT80 plays an important role in osteogenesis through regulating Hedgehog/Gli signal pathways. PMID:22771375

  20. Study of clathrate hydrates via equilibrium molecular-dynamics simulation employing polarisable and non-polarisable, rigid and flexible water models.

    PubMed

    Burnham, Christian J; English, Niall J

    2016-04-28

    Equilibrium molecular-dynamics (MD) simulations have been performed on metastable sI and sII polymorphs of empty hydrate lattices, in addition to liquid water and ice Ih. The non-polarisable TIP4P-2005, simple point charge model (SPC), and polarisable Thole-type models (TTM): TTM2, TTM3, and TTM4 water models were used in order to survey the differences between models and to see what differences can be expected when polarisability is incorporated. Rigid and flexible variants were used of each model to gauge the effects of flexibility. Power spectra are calculated and compared to density-of-states spectra inferred from inelastic neutron scattering (INS) measurements. Thermodynamic properties were also calculated, as well as molecular-dipole distributions. It was concluded that TTM models offer optimal fidelity vis-à-vis INS spectra, together with thermodynamic properties, with the flexible TTM2 model offering optimal placement of vibrational modes. PMID:27131553

  1. Study of clathrate hydrates via equilibrium molecular-dynamics simulation employing polarisable and non-polarisable, rigid and flexible water models

    NASA Astrophysics Data System (ADS)

    Burnham, Christian J.; English, Niall J.

    2016-04-01

    Equilibrium molecular-dynamics (MD) simulations have been performed on metastable sI and sII polymorphs of empty hydrate lattices, in addition to liquid water and ice Ih. The non-polarisable TIP4P-2005, simple point charge model (SPC), and polarisable Thole-type models (TTM): TTM2, TTM3, and TTM4 water models were used in order to survey the differences between models and to see what differences can be expected when polarisability is incorporated. Rigid and flexible variants were used of each model to gauge the effects of flexibility. Power spectra are calculated and compared to density-of-states spectra inferred from inelastic neutron scattering (INS) measurements. Thermodynamic properties were also calculated, as well as molecular-dipole distributions. It was concluded that TTM models offer optimal fidelity vis-à-vis INS spectra, together with thermodynamic properties, with the flexible TTM2 model offering optimal placement of vibrational modes.

  2. A polarisation modulation scheme for measuring vacuum magnetic birefringence with static fields

    NASA Astrophysics Data System (ADS)

    Zavattini, G.; Della Valle, F.; Ejlli, A.; Ruoso, G.

    2016-05-01

    A novel polarisation modulation scheme for polarimeters based on Fabry-Perot cavities is presented. The application to the measurement of the magnetic birefringence of vacuum with the HERA superconducting magnets in the ALPS-II configuration is discussed.

  3. Compton scattering and nucleon polarisabilities in chiral EFT: Status and future

    NASA Astrophysics Data System (ADS)

    Grießhammer, Harald W.; McGovern, Judith A.; Phillips, Daniel R.

    2016-05-01

    We review theoretical progress and prospects for determining the nucleon's static dipole polarisabilities from Compton scattering on few-nucleon targets, including new values; see Refs. [1-5] for details and a more thorough bibliography.

  4. Spin echo small angle neutron scattering using a continuously pumped {sup 3}He neutron polarisation analyser

    SciTech Connect

    Parnell, S. R.; Li, K.; Yan, H.; Stonaha, P.; Li, F.; Wang, T.; Baxter, D. V.; Snow, W. M.; Washington, A. L.; Walsh, A.; Chen, W. C.; Parnell, A. J.; Fairclough, J. P. A.; Pynn, R.

    2015-02-15

    We present a new instrument for spin echo small angle neutron scattering (SESANS) developed at the Low Energy Neutron Source at Indiana University. A description of the various instrument components is given along with the performance of these components. At the heart of the instrument are a series of resistive coils to encode the neutron trajectory into the neutron polarisation. These are shown to work well over a broad range of neutron wavelengths. Neutron polarisation analysis is accomplished using a continuously operating neutron spin filter polarised by Rb spin-exchange optical pumping of {sup 3}He. We describe the performance of the analyser along with a study of the {sup 3}He polarisation stability and its implications for SESANS measurements. Scattering from silica Stöber particles is investigated and agrees with samples run on similar instruments.

  5. Enteric neurons show a primary cilium

    PubMed Central

    Luesma, Mª José; Cantarero, Irene; Castiella, Tomás; Soriano, Mario; Garcia–Verdugo, José Manuel; Junquera, Concepción

    2013-01-01

    The primary cilium is a non-motile cilium whose structure is 9+0. It is involved in co-ordinating cellular signal transduction pathways, developmental processes and tissue homeostasis. Defects in the structure or function of the primary cilium underlie numerous human diseases, collectively termed ciliopathies. The presence of single cilia in the central nervous system (CNS) is well documented, including some choroid plexus cells, neural stem cells, neurons and astrocytes, but the presence of primary cilia in differentiated neurons of the enteric nervous system (ENS) has not yet been described in mammals to the best of our knowledge. The enteric nervous system closely resembles the central nervous system. In fact, the ultrastructure of the ENS is more similar to the CNS ultrastructure than to the rest of the peripheral nervous system. This research work describes for the first time the ultrastructural characteristics of the single cilium in neurons of rat duodenum myenteric plexus, and reviews the cilium function in the CNS to propose the possible role of cilia in the ENS cells. PMID:23205631

  6. Primary ciliary dyskinesia.

    PubMed Central

    Le Mauviel, L.

    1991-01-01

    Primary ciliary dyskinesia represents a group of heritable disorders of cilia and sperm affecting between 1 in 15,000 and 1 in 30,000 persons. Those affected lack measurable mucociliary clearance and suffer the constant misery of rhinorrhea and chronic productive cough. Because mucociliary clearance constitutes one of the respiratory system's major lines of defense, these patients are vulnerable to chronic sinusitis, bronchitis, pneumonia, and otitis media. Left untreated, these problems may progress to bronchiectasis, found frequently in adult patients, or pulmonary hypertension with eventual cor pulmonale. Screening for this disorder includes some simple and inexpensive methods as well as more exotic techniques requiring special camera equipment and an electron microscope to make a definitive diagnosis. Physiotherapy techniques can be taught to patients with primary ciliary dyskinesia and go a long way toward making up for the lack of mucociliary clearance. Vigorous bronchopulmonary toilet and palliative measures may enable these patients to enjoy relatively normal lives. PMID:1949776

  7. High resolution VLBI polarisation imaging of AGN with the Maximum Entropy Method

    NASA Astrophysics Data System (ADS)

    Coughlan, Colm P.; Gabuzda, Denise C.

    2016-08-01

    Radio polarisation images of the jets of Active Galactic Nuclei (AGN) can provide a deep insight into the launching and collimation mechanisms of relativistic jets. However, even at VLBI scales, resolution is often a limiting factor in the conclusions that can be drawn from observations. The Maximum Entropy Method (MEM) is a deconvolution algorithm that can outperform the more common CLEAN algorithm in many cases, particularly when investigating structures present on scales comparable to or smaller than the nominal beam size with "super-resolution". A new implementation of the MEM suitable for single- or multiple-wavelength VLBI polarisation observations has been developed and is described here. Monte Carlo simulations comparing the performances of CLEAN and MEM at reconstructing the properties of model images are presented; these demonstrate the enhanced reliability of MEM over CLEAN when images of the fractional polarisation and polarisation angle are constructed using convolving beams that are appreciably smaller than the full CLEAN beam. The results of using this new MEM software to image VLBA observations of the AGN 0716+714 at six different wavelengths are presented, and compared to corresponding maps obtained with CLEAN. MEM and CLEAN maps of Stokes I, the polarised flux, the fractional polarisation and the polarisation angle are compared for convolving beams ranging from the full CLEAN beam down to a beam one-third of this size. MEM's ability to provide more trustworthy polarisation imaging than a standard CLEAN-based deconvolution when convolving beams appreciably smaller than the full CLEAN beam are used is discussed.

  8. LEKIDs as mm-Wave Polarisation Analysers: Fabrication, Test Bench and Early Results

    NASA Astrophysics Data System (ADS)

    Tartari, A.; Bélier, B.; Bleurvacq, N.; Calvo, M.; Cammilleri, D.; Decourcelle, T.; Monfardini, A.; Moric, I.; Piat, M.; Prêle, D.; Smoot, G. F.

    2016-07-01

    We have demonstrated in an earlier paper that LEKIDs can be used in a polarisation selective way in a filled array configuration. A polarised response can be achieved by means of thick Nb polarising grids lithographed on the rear side of a 300 microns silicon wafer, on which Al resonators have been previously patterned. In the most interesting scheme that we have investigated, a unit cell formed by 4 pixels (2 by 2) responds simultaneously to two orthogonal (cartesian) polarisation states. To assess the effectiveness of this detection scheme, we have fabricated a first generation of devices (9 small arrays, 20-25 pixels each, on a 4^' ' } Silicon wafer) by using a double-sided mask aligner suitable for a precise positioning of the individual grids in correspondence of each resonator's meander, for the different LEKID geometries. We describe here the realisation of these first devices. The construction of a dedicated polarimetric test bench is also described in this contribution, together with the first characterisation results. We consider this activity as a first and necessary step to evaluate the polarisation purity attainable with polarisation-sensitive pixels whose size is comparable to the wavelength. This is a fundamental information to drive further studies.

  9. Polarisation response of delay dependent absorption modulation in strong field dressed helium atoms probed near threshold

    NASA Astrophysics Data System (ADS)

    Simpson, E. R.; Sanchez-Gonzalez, A.; Austin, D. R.; Diveki, Z.; Hutchinson, S. E. E.; Siegel, T.; Ruberti, M.; Averbukh, V.; Miseikis, L.; Strüber, C. S.; Chipperfield, L.; Marangos, J. P.

    2016-08-01

    We present the first measurement of the vectorial response of strongly dressed helium atoms probed by an attosecond pulse train (APT) polarised either parallel or perpendicular to the dressing field polarisation. The transient absorption is probed as a function of delay between the APT and the linearly polarised 800 nm field of peak intensity 1.3× {10}14 {{W}} {{cm}}-2. The APT spans the photon energy range 16–42 eV, covering the first ionisation energy of helium (24.59 eV). With parallel polarised dressing and probing fields, we observe modulations with periods of one half and one quarter of the dressing field period. When the polarisation of the dressing field is altered from parallel to perpendicular with respect to the APT polarisation we observe a large suppression in the modulation depth of the above ionisation threshold absorption. In addition to this we present the intensity dependence of the harmonic modulation depth as a function of delay between the dressing and probe fields, with dressing field peak intensities ranging from 2 × 1012 to 2 × 1014 {{W}} {{cm}}-2. We compare our experimental results with a full-dimensional solution of the single-atom time-dependent (TD) Schrödinger equation obtained using the recently developed abinitio TD B-spline ADC method and find good qualitative agreement for the above threshold harmonics.

  10. LEKIDs as mm-Wave Polarisation Analysers: Fabrication, Test Bench and Early Results

    NASA Astrophysics Data System (ADS)

    Tartari, A.; Bélier, B.; Bleurvacq, N.; Calvo, M.; Cammilleri, D.; Decourcelle, T.; Monfardini, A.; Moric, I.; Piat, M.; Prêle, D.; Smoot, G. F.

    2015-12-01

    We have demonstrated in an earlier paper that LEKIDs can be used in a polarisation selective way in a filled array configuration. A polarised response can be achieved by means of thick Nb polarising grids lithographed on the rear side of a 300 microns silicon wafer, on which Al resonators have been previously patterned. In the most interesting scheme that we have investigated, a unit cell formed by 4 pixels (2 by 2) responds simultaneously to two orthogonal (cartesian) polarisation states. To assess the effectiveness of this detection scheme, we have fabricated a first generation of devices (9 small arrays, 20-25 pixels each, on a 4^' ' } Silicon wafer) by using a double-sided mask aligner suitable for a precise positioning of the individual grids in correspondence of each resonator's meander, for the different LEKID geometries. We describe here the realisation of these first devices. The construction of a dedicated polarimetric test bench is also described in this contribution, together with the first characterisation results. We consider this activity as a first and necessary step to evaluate the polarisation purity attainable with polarisation-sensitive pixels whose size is comparable to the wavelength. This is a fundamental information to drive further studies.

  11. Structural and energetic effects of the use of polarisable water to solvate proteins

    NASA Astrophysics Data System (ADS)

    Bachmann, Stephan J.; van Gunsteren, Wilfred F.

    2015-09-01

    Using a non-polarisable model (simple-point-charge (SPC)) for liquid water and two polarisable water models (COS/G2, COS/D), the effect of introducing molecular polarisability into the solvent upon protein structure and energetics is investigated for eight proteins, hen egg-white lysozyme (HEWL), major cold shock protein (CspA), protein G (GP), chorismate mutase (CM), the C-terminal domain of the ribosomal protein L7/L12 (RB), the amino terminal domain of phage 434 repressor (GRP), a 12-residue β-hairpin (DNV) and the GCN trigger peptide (GTP), using MD simulation, one 50 ns simulation and four additional 20 ns simulations for each protein and each water model. The differences in overall structural and energetic properties of the proteins induced by the three different water models are small, except for the amino-terminal domain of phage 434 repressor (GRP). The polarisable COS/G2 water model induces a slightly stronger interaction with the proteins modelled using the GROMOS 54A7 force field than the non-polarisable SPC water model, while for the polarisable COS/D water model the opposite effect is observed.

  12. Polarised skylight and the landmark panorama provide night-active bull ants with compass information during route following.

    PubMed

    Reid, Samuel F; Narendra, Ajay; Hemmi, Jan M; Zeil, Jochen

    2011-02-01

    Navigating animals are known to use a number of celestial and terrestrial compass cues that allow them to determine and control their direction of travel. Which of the cues dominate appears to depend on their salience. Here we show that night-active bull ants attend to both the pattern of polarised skylight and the landmark panorama in their familiar habitat. When the two directional cues are in conflict, ants choose a compromise direction. However, landmark guidance appears to be the primary mechanism of navigation used by forager ants, with those cues in the direction of heading having the greatest influence on navigation. Different colonies respond to the removal of these cues to different degrees, depending on the directional information provided by the local landmark panorama. Interestingly, other parts of the surrounding panorama also influence foraging speed and accuracy, suggesting that they too play a role in navigation. PMID:21228195

  13. Computational models of the primary cilium and endothelial mechanotransmission.

    PubMed

    Lim, Yi Chung; Cooling, Michael T; Long, David S

    2015-06-01

    In endothelial cells (ECs), the mechanotransduction of fluid shear stress is partially dependent on the transmission of force from the fluid into the cell (mechanotransmission). The role of the primary cilium in EC mechanotransmission is not yet known. To motivate a framework towards quantifying cilia contribution to EC mechanotransmission, we have reviewed mechanical models of both (1) the primary cilium (three-dimensional and lower-dimensional) and (2) whole ECs (finite element, non-finite element, and tensegrity). Both the primary cilia and whole EC models typically incorporate fluid-induced wall shear stress and spatial geometry based on experimentally acquired images of cells. This paper presents future modelling directions as well as the major goals towards integrating primary cilium models into a multi-component EC mechanical model. Finally, we outline how an integrated cilium-EC model can be used to better understand mechanotransduction in the endothelium. PMID:25366114

  14. Synergistic activity of polarised osteoblasts inside condensations cause their differentiation

    PubMed Central

    Kaul, Himanshu; Hall, Brian K.; Newby, Chris; Ventikos, Yiannis

    2015-01-01

    Condensation of pre-osteogenic, or pre-chondrogenic, cells is the first of a series of processes that initiate skeletal development. We present a validated, novel, three-dimensional agent-based model of in vitro intramembranous osteogenic condensation. The model, informed by system heterogeneity and relying on an interaction-reliant strategy, is shown to be sensitive to ‘rules’ capturing condensation growth and can be employed to track activity of individual cells to observe their macroscopic impact. It, therefore, makes available previously inaccessible data, offering new insights and providing a new context for exploring the emergence, as well as normal and abnormal development, of osteogenic structures. Of the several stages of condensation we investigate osteoblast ‘burial’ within the osteoid they deposit. The mechanisms underlying entrapment – required for osteoblasts to differentiate into osteocytes – remain a matter of conjecture with several hypotheses claiming to capture this important transition. Computational examination of this transition indicates that osteoblasts neither turn off nor slow down their matrix secreting genes – a widely held view; nor do they secrete matrix randomly. The model further reveals that osteoblasts display polarised behaviour to deposit osteoid. This is both an important addition to our understanding of condensation and an important validation of the model’s utility. PMID:26146365

  15. Exclusive ρ0 muoproduction on transversely polarised protons and deuterons

    NASA Astrophysics Data System (ADS)

    Adolph, C.; Alekseev, M. G.; Alexakhin, V. Yu.; Alexandrov, Yu.; Alexeev, G. D.; Amoroso, A.; Antonov, A. A.; Austregesilo, A.; Badełek, B.; Balestra, F.; Barth, J.; Baum, G.; Bedfer, Y.; Bernhard, J.; Bertini, R.; Bettinelli, M.; Bicker, K.; Bieling, J.; Birsa, R.; Bisplinghoff, J.; Bordalo, P.; Bradamante, F.; Braun, C.; Bravar, A.; Bressan, A.; Büchele, M.; Burtin, E.; Capozza, L.; Chiosso, M.; Chung, S. U.; Cicuttin, A.; Crespo, M. L.; Dalla Torre, S.; Das, S.; Dasgupta, S. S.; Dasgupta, S.; Denisov, O. Yu.; Dhara, L.; Donskov, S. V.; Doshita, N.; Duic, V.; Dünnweber, W.; Dziewiecki, M.; Efremov, A.; Elia, C.; Eversheim, P. D.; Eyrich, W.; Faessler, M.; Ferrero, A.; Filin, A.; Finger, M.; Finger, M.; Fischer, H.; Franco, C.; du Fresne von Hohenesche, N.; Friedrich, J. M.; Frolov, V.; Garfagnini, R.; Gautheron, F.; Gavrichtchouk, O. P.; Gerassimov, S.; Geyer, R.; Giorgi, M.; Gnesi, I.; Gobbo, B.; Goertz, S.; Grabmüller, S.; Grasso, A.; Grube, B.; Gushterski, R.; Guskov, A.; Guthörl, T.; Haas, F.; von Harrach, D.; Heinsius, F. H.; Herrmann, F.; Heß, C.; Hinterberger, F.; Horikawa, N.; Höppner, Ch.; d'Hose, N.; Ishimoto, S.; Ivanov, O.; Ivanshin, Yu.; Iwata, T.; Jahn, R.; Jary, V.; Jasinski, P.; Jegou, G.; Joosten, R.; Kabuß, E.; Kang, D.; Ketzer, B.; Khaustov, G. V.; Khokhlov, Yu. A.; Kisselev, Yu.; Klein, F.; Klimaszewski, K.; Koblitz, S.; Koivuniemi, J. H.; Kolosov, V. N.; Kondo, K.; Königsmann, K.; Konorov, I.; Konstantinov, V. F.; Korzenev, A.; Kotzinian, A. M.; Kouznetsov, O.; Krämer, M.; Kroumchtein, Z. V.; Kunne, F.; Kurek, K.; Lauser, L.; Lednev, A. A.; Lehmann, A.; Levorato, S.; Lichtenstadt, J.; Liska, T.; Maggiora, A.; Magnon, A.; Makke, N.; Mallot, G. K.; Mann, A.; Marchand, C.; Martin, A.; Marzec, J.; Matsuda, T.; Meshcheryakov, G.; Meyer, W.; Michigami, T.; Mikhailov, Yu. V.; Moinester, M. A.; Morreale, A.; Mutter, A.; Nagaytsev, A.; Nagel, T.; Negrini, T.; Nerling, F.; Neubert, S.; Neyret, D.; Nikolaenko, V. I.; Nowak, W.-D.; Nunes, A. S.; Olshevsky, A. G.; Ostrick, M.; Padee, A.; Panknin, R.; Panzieri, D.; Parsamyan, B.; Paul, S.; Perevalova, E.; Pesaro, G.; Peshekhonov, D. V.; Piragino, G.; Platchkov, S.; Pochodzalla, J.; Polak, J.; Polyakov, V. A.; Pretz, J.; Quaresma, M.; Quintans, C.; Rajotte, J.-F.; Ramos, S.; Rapatsky, V.; Reicherz, G.; Richter, A.; Rocco, E.; Rondio, E.; Rossiyskaya, N. S.; Ryabchikov, D. I.; Samoylenko, V. D.; Sandacz, A.; Sapozhnikov, M. G.; Sarkar, S.; Savin, I. A.; Sbrizzai, G.; Schiavon, P.; Schill, C.; Schlüter, T.; Schmidt, K.; Schmitt, L.; Schönning, K.; Schopferer, S.; Schott, M.; Schröder, W.; Shevchenko, O. Yu.; Silva, L.; Sinha, L.; Sissakian, A. N.; Slunecka, M.; Smirnov, G. I.; Sosio, S.; Sozzi, F.; Srnka, A.; Steiger, L.; Stolarski, M.; Sulc, M.; Sulej, R.; Suzuki, H.; Sznajder, P.; Takekawa, S.; Ter Wolbeek, J.; Tessaro, S.; Tessarotto, F.; Tkatchev, L. G.; Uhl, S.; Uman, I.; Vandenbroucke, M.; Virius, M.; Vlassov, N. V.; Wang, L.; Wilfert, M.; Windmolders, R.; Wiślicki, W.; Wollny, H.; Zaremba, K.; Zavertyaev, M.; Zemlyanichkina, E.; Ziembicki, M.; Zhuravlev, N.; Zvyagin, A.

    2012-12-01

    The transverse target spin azimuthal asymmetry AUTsin(ϕ-ϕS) in hard exclusive production of ρ0 mesons was measured at COMPASS by scattering 160 GeV/c muons off transversely polarised protons and deuterons. The measured asymmetry is sensitive to the nucleon helicity-flip generalised parton distributions Eq, which are related to the orbital angular momentum of quarks in the nucleon. The Q2, xBj and pT2 dependence of AUTsin(ϕ-ϕS) is presented in a wide kinematic range: 1 (

  16. Debye decomposition of time-lapse spectral induced polarisation data

    NASA Astrophysics Data System (ADS)

    Weigand, M.; Kemna, A.

    2016-01-01

    Spectral induced polarisation (SIP) measurements capture the low-frequency electrical properties of soils and rocks and provide a non-invasive means to access lithological, hydrogeological, and geochemical properties of the subsurface. The Debye decomposition (DD) approach is now increasingly being used to analyse SIP signatures in terms of relaxation time distributions due to its flexibility regarding the shape of the spectra. Imaging and time-lapse (monitoring) SIP measurements, capturing SIP variations in space and time, respectively, are now more and more conducted and lead to a drastic increase in the number of spectra considered, which prompts the need for robust and reliable DD tools to extract quantitative parameters from such data. We here present an implementation of the DD method for the analysis of a series of SIP data sets which are expected to only smoothly change in terms of spectral behaviour, such as encountered in many time-lapse applications where measurement geometry does not change. The routine is based on a non-linear least-squares inversion scheme with smoothness constraints on the spectral variation and in addition from one spectrum of the series to the next to deal with the inherent ill-posedness and non-uniqueness of the problem. By means of synthetic examples with typical SIP characteristics we elucidate the influence of the number and range of considered relaxation times on the inversion results. The source code of the presented routines is provided under an open source licence as a basis for further applications and developments.

  17. The Zebrafish Orthologue of the Dyslexia Candidate Gene DYX1C1 Is Essential for Cilia Growth and Function

    PubMed Central

    Chandrasekar, Gayathri; Vesterlund, Liselotte; Hultenby, Kjell; Tapia-Páez, Isabel; Kere, Juha

    2013-01-01

    DYX1C1, a susceptibility gene for dyslexia, encodes a tetratricopeptide repeat domain containing protein that has been implicated in neuronal migration in rodent models. The developmental role of this gene remains unexplored. To understand the biological function(s) of zebrafish dyx1c1 during embryonic development, we cloned the zebrafish dyx1c1 and used morpholino-based knockdown strategy. Quantitative real-time PCR analysis revealed the presence of dyx1c1 transcripts in embryos, early larval stages and in a wide range of adult tissues. Using mRNA in situ hybridization, we show here that dyx1c1 is expressed in many ciliated tissues in zebrafish. Inhibition of dyx1c1 produced pleiotropic phenotypes characteristically associated with cilia defects such as body curvature, hydrocephalus, situs inversus and kidney cysts. We also demonstrate that in dyx1c1 morphants, cilia length is reduced in several organs including Kupffer’s vesicle, pronephros, spinal canal and olfactory placode. Furthermore, electron microscopic analysis of cilia in dyx1c1 morphants revealed loss of both outer (ODA) and inner dynein arms (IDA) that have been shown to be required for cilia motility. Considering all these results, we propose an essential role for dyx1c1 in cilia growth and function. PMID:23650548

  18. Regulation of Chlamydomonas flagella and ependymal cell motile cilia by ceramide-mediated translocation of GSK3

    PubMed Central

    Kong, Ji Na; Hardin, Kara; Dinkins, Michael; Wang, Guanghu; He, Qian; Mujadzic, Tarik; Zhu, Gu; Bielawski, Jacek; Spassieva, Stefka; Bieberich, Erhard

    2015-01-01

    Cilia are important organelles formed by cell membrane protrusions; however, little is known about their regulation by membrane lipids. We characterize a novel activation mechanism for glycogen synthase kinase-3 (GSK3) by the sphingolipids phytoceramide and ceramide that is critical for ciliogenesis in Chlamydomonas and murine ependymal cells, respectively. We show for the first time that Chlamydomonas expresses serine palmitoyl transferase (SPT), the first enzyme in (phyto)ceramide biosynthesis. Inhibition of SPT in Chlamydomonas by myriocin led to loss of flagella and reduced tubulin acetylation, which was prevented by supplementation with the precursor dihydrosphingosine. Immunocytochemistry showed that (phyto)ceramide was colocalized with phospho–Tyr-216-GSK3 (pYGSK3) at the base and tip of Chlamydomonas flagella and motile cilia in ependymal cells. The (phyto)ceramide distribution was consistent with that of a bifunctional ceramide analogue UV cross-linked and visualized by click-chemistry–mediated fluorescent labeling. Ceramide depletion, by myriocin or neutral sphingomyelinase deficiency (fro/fro mouse), led to GSK3 dephosphorylation and defective flagella and cilia. Motile cilia were rescued and pYGSK3 localization restored by incubation of fro/fro ependymal cells with exogenous C24:1 ceramide, which directly bound to pYGSK3. Our findings suggest that (phyto)ceramide-mediated translocation of pYGSK into flagella and cilia is an evolutionarily conserved mechanism fundamental to the regulation of ciliogenesis. PMID:26446842

  19. The Intraflagellar Transport Protein IFT27 Promotes BBSome Exit from Cilia through the GTPase ARL6/BBS3

    PubMed Central

    Liew, Gerald M.; Ye, Fan; Nager, Andrew R.; Murphy, J. Patrick; Lee, Jaclyn S.; Aguiar, Mike; Breslow, David K.; Gygi, Steven P.; Nachury, Maxence V.

    2014-01-01

    SUMMARY The sorting of signaling receptors into and out of cilia relies on the BBSome, a complex of Bardet-Biedl syndrome (BBS) proteins, and on the intraflagellar transport (IFT) machinery. GTP loading onto the Arf-like GTPase ARL6/BBS3 drives assembly of a membrane-apposed BBSome coat that promotes cargo entry into cilia, yet how and where ARL6 is activated remains elusive. Here, we show that the Rab-like GTPase IFT27/RABL4, a known component of IFT complex B, promotes the exit of BBSome and associated cargoes from cilia. Unbiased proteomics and biochemical reconstitution assays show that, upon disengagement from the rest of IFT-B, IFT27 directly interacts with the nucleotide-free form of ARL6. Furthermore, IFT27 prevents aggregation of nucleotide-free ARL6 in solution. Thus, we propose that IFT27 separates from IFT-B inside cilia to promote ARL6 activation, BBSome coat assembly and subsequent ciliary exit, mirroring the process by which BBSome mediates cargo entry into cilia. PMID:25443296

  20. Functional optical coherence tomography for high-resolution mapping of cilia beat frequency in the mouse oviduct in vivo

    NASA Astrophysics Data System (ADS)

    Wang, Shang; Burton, Jason C.; Behringer, Richard R.; Larina, Irina V.

    2016-02-01

    Since mouse is a superior model for genetic analysis of human disorders, reproductive studies in mice have significant implications on further understanding of fertility and infertility in humans. Fertilized oocytes are transported through the reproductive tract by motile cilia lining the lumen of the oviduct as well as by oviduct contractions. While the role of cilia is well recognized, ciliary dynamics in the oviduct is not well understood, largely owing to the lack of live imaging approaches. Here, we report in vivo micro-scale mapping of cilia and cilia beat frequency (CBF) in the mouse oviduct using optical coherence tomography (OCT). This functional imaging method is based on spectral analysis of the OCT speckle variations produced by the beat of cilia in the oviduct, which does not require exogenous contrast agents. Animal procedures similar to the ones used for production of transgenic mice are utilized to expose the reproductive organs for imaging in anesthetized females. In this paper, we first present in vivo structural imaging of the mouse oviduct capturing the oocyte and the preimplantation embryo and then show the result of depth-resolved high-resolution CBF mapping in the ampulla of the live mouse. These data indicate that this structural and functional OCT imaging approach can be a useful tool for a variety of live investigations of mammalian reproduction and infertility.

  1. DNAH6 and Its Interactions with PCD Genes in Heterotaxy and Primary Ciliary Dyskinesia.

    PubMed

    Li, You; Yagi, Hisato; Onuoha, Ezenwa Obi; Damerla, Rama Rao; Francis, Richard; Furutani, Yoshiyuki; Tariq, Muhammad; King, Stephen M; Hendricks, Gregory; Cui, Cheng; Saydmohammed, Manush; Lee, Dong Min; Zahid, Maliha; Sami, Iman; Leatherbury, Linda; Pazour, Gregory J; Ware, Stephanie M; Nakanishi, Toshio; Goldmuntz, Elizabeth; Tsang, Michael; Lo, Cecilia W

    2016-02-01

    Heterotaxy, a birth defect involving left-right patterning defects, and primary ciliary dyskinesia (PCD), a sinopulmonary disease with dyskinetic/immotile cilia in the airway are seemingly disparate diseases. However, they have an overlapping genetic etiology involving mutations in cilia genes, a reflection of the common requirement for motile cilia in left-right patterning and airway clearance. While PCD is a monogenic recessive disorder, heterotaxy has a more complex, largely non-monogenic etiology. In this study, we show mutations in the novel dynein gene DNAH6 can cause heterotaxy and ciliary dysfunction similar to PCD. We provide the first evidence that trans-heterozygous interactions between DNAH6 and other PCD genes potentially can cause heterotaxy. DNAH6 was initially identified as a candidate heterotaxy/PCD gene by filtering exome-sequencing data from 25 heterotaxy patients stratified by whether they have airway motile cilia defects. dnah6 morpholino knockdown in zebrafish disrupted motile cilia in Kupffer's vesicle required for left-right patterning and caused heterotaxy with abnormal cardiac/gut looping. Similarly DNAH6 shRNA knockdown disrupted motile cilia in human and mouse respiratory epithelia. Notably a heterotaxy patient harboring heterozygous DNAH6 mutation was identified to also carry a rare heterozygous PCD-causing DNAI1 mutation, suggesting a DNAH6/DNAI1 trans-heterozygous interaction. Furthermore, sequencing of 149 additional heterotaxy patients showed 5 of 6 patients with heterozygous DNAH6 mutations also had heterozygous mutations in DNAH5 or other PCD genes. We functionally assayed for DNAH6/DNAH5 and DNAH6/DNAI1 trans-heterozygous interactions using subthreshold double-morpholino knockdown in zebrafish and showed this caused heterotaxy. Similarly, subthreshold siRNA knockdown of Dnah6 in heterozygous Dnah5 or Dnai1 mutant mouse respiratory epithelia disrupted motile cilia function. Together, these findings support an oligogenic disease

  2. DNAH6 and Its Interactions with PCD Genes in Heterotaxy and Primary Ciliary Dyskinesia

    PubMed Central

    Onuoha, Ezenwa Obi; Damerla, Rama Rao; Francis, Richard; Furutani, Yoshiyuki; Tariq, Muhammad; King, Stephen M.; Hendricks, Gregory; Cui, Cheng; Saydmohammed, Manush; Lee, Dong Min; Zahid, Maliha; Sami, Iman; Leatherbury, Linda; Pazour, Gregory J.; Ware, Stephanie M.; Nakanishi, Toshio; Goldmuntz, Elizabeth; Tsang, Michael; Lo, Cecilia W.

    2016-01-01

    Heterotaxy, a birth defect involving left-right patterning defects, and primary ciliary dyskinesia (PCD), a sinopulmonary disease with dyskinetic/immotile cilia in the airway are seemingly disparate diseases. However, they have an overlapping genetic etiology involving mutations in cilia genes, a reflection of the common requirement for motile cilia in left-right patterning and airway clearance. While PCD is a monogenic recessive disorder, heterotaxy has a more complex, largely non-monogenic etiology. In this study, we show mutations in the novel dynein gene DNAH6 can cause heterotaxy and ciliary dysfunction similar to PCD. We provide the first evidence that trans-heterozygous interactions between DNAH6 and other PCD genes potentially can cause heterotaxy. DNAH6 was initially identified as a candidate heterotaxy/PCD gene by filtering exome-sequencing data from 25 heterotaxy patients stratified by whether they have airway motile cilia defects. dnah6 morpholino knockdown in zebrafish disrupted motile cilia in Kupffer’s vesicle required for left-right patterning and caused heterotaxy with abnormal cardiac/gut looping. Similarly DNAH6 shRNA knockdown disrupted motile cilia in human and mouse respiratory epithelia. Notably a heterotaxy patient harboring heterozygous DNAH6 mutation was identified to also carry a rare heterozygous PCD-causing DNAI1 mutation, suggesting a DNAH6/DNAI1 trans-heterozygous interaction. Furthermore, sequencing of 149 additional heterotaxy patients showed 5 of 6 patients with heterozygous DNAH6 mutations also had heterozygous mutations in DNAH5 or other PCD genes. We functionally assayed for DNAH6/DNAH5 and DNAH6/DNAI1 trans-heterozygous interactions using subthreshold double-morpholino knockdown in zebrafish and showed this caused heterotaxy. Similarly, subthreshold siRNA knockdown of Dnah6 in heterozygous Dnah5 or Dnai1 mutant mouse respiratory epithelia disrupted motile cilia function. Together, these findings support an oligogenic disease

  3. Polarised photoselection and molecular dynamics in liquid crystals and proteins

    NASA Astrophysics Data System (ADS)

    Bryant, Jason

    Time resolved fluorescence polarisation studies of probe motion in a liquid crystal and protein matrix are presented. In this work, the dynamics and orientational properties of four common laser dyes (Oxazine 1, Oxazine 4, Rhodamine 6G and Rhodamine B) are determined in the liquid crystal 5CB. These studies provide the first direct measurement of θ and φ diffusion (hitherto unobserved) of molecular probes in a nematic host. A distinct anisotropy in θ and φ motion is observed, θ diffusion dynamics show a conventional Arrhenius temperature dependence in the approach to T NI. In contrast, φ motion is largely temperature independent and shows some evidence of slowing in the vicinity of the phase transition. From constructing the orientational distribution function P(θ), it is revealed that the dyes are aligned in pockets between the flexible tails of the host (θ ~ 38°) and conventional models assuming a cylindrical potential are shown to be non-applicable. These effects are most pronounced in Oxazine 4 whose alignment correlates strongly with that of the alkyl tails in 5CB. Here θ and φ diffusion dynamics are highly anisotropic with the ratio of τ20/τ22 in the region of 5:1. In the approach to TNI τ22 is seen to increase by c.a. 50% whilst τ20 shows a c.a 70% decrease. Measurements of Oxazine 4 motion in the Isotropic phase of 5CB indicate that the dye remains strongly correlated with the liquid crystal. Two diffusion times are observed consistent with restricted rotational diffusion within a more slowly diffusing arrangement (domain) of solvent molecules. The domain motion is seen to exhibit a Landau-de Gennes type temperature dependence whilst probe motion within the domains is seen to be largely temperature independent, similar behaviour having been recently reported for diffusion in pure 5CB. A central portion of the work concerns the development of a wholly new approach to polarised photoselection. A novel 3 beam photoselection technique in which

  4. Genetic Analysis Reveals a Hierarchy of Interactions between Polycystin-Encoding Genes and Genes Controlling Cilia Function during Left-Right Determination

    PubMed Central

    Grimes, Daniel T.; Keynton, Jennifer L.; Buenavista, Maria T.; Jin, Xingjian; Patel, Saloni H.; Kyosuke, Shinohara; Williams, Debbie J.; Hamada, Hiroshi; Hussain, Rohanah; Nauli, Surya M.; Norris, Dominic P.

    2016-01-01

    During mammalian development, left-right (L-R) asymmetry is established by a cilia-driven leftward fluid flow within a midline embryonic cavity called the node. This ‘nodal flow’ is detected by peripherally-located crown cells that each assemble a primary cilium which contain the putative Ca2+ channel PKD2. The interaction of flow and crown cell cilia promotes left side-specific expression of Nodal in the lateral plate mesoderm (LPM). Whilst the PKD2-interacting protein PKD1L1 has also been implicated in L-R patterning, the underlying mechanism by which flow is detected and the genetic relationship between Polycystin function and asymmetric gene expression remains unknown. Here, we characterize a Pkd1l1 mutant line in which Nodal is activated bilaterally, suggesting that PKD1L1 is not required for LPM Nodal pathway activation per se, but rather to restrict Nodal to the left side downstream of nodal flow. Epistasis analysis shows that Pkd1l1 acts as an upstream genetic repressor of Pkd2. This study therefore provides a genetic pathway for the early stages of L-R determination. Moreover, using a system in which cultured cells are supplied artificial flow, we demonstrate that PKD1L1 is sufficient to mediate a Ca2+ signaling response after flow stimulation. Finally, we show that an extracellular PKD domain within PKD1L1 is crucial for PKD1L1 function; as such, destabilizing the domain causes L-R defects in the mouse. Our demonstration that PKD1L1 protein can mediate a response to flow coheres with a mechanosensation model of flow sensation in which the force of fluid flow drives asymmetric gene expression in the embryo. PMID:27272319

  5. Genetic Analysis Reveals a Hierarchy of Interactions between Polycystin-Encoding Genes and Genes Controlling Cilia Function during Left-Right Determination.

    PubMed

    Grimes, Daniel T; Keynton, Jennifer L; Buenavista, Maria T; Jin, Xingjian; Patel, Saloni H; Kyosuke, Shinohara; Vibert, Jennifer; Williams, Debbie J; Hamada, Hiroshi; Hussain, Rohanah; Nauli, Surya M; Norris, Dominic P

    2016-06-01

    During mammalian development, left-right (L-R) asymmetry is established by a cilia-driven leftward fluid flow within a midline embryonic cavity called the node. This 'nodal flow' is detected by peripherally-located crown cells that each assemble a primary cilium which contain the putative Ca2+ channel PKD2. The interaction of flow and crown cell cilia promotes left side-specific expression of Nodal in the lateral plate mesoderm (LPM). Whilst the PKD2-interacting protein PKD1L1 has also been implicated in L-R patterning, the underlying mechanism by which flow is detected and the genetic relationship between Polycystin function and asymmetric gene expression remains unknown. Here, we characterize a Pkd1l1 mutant line in which Nodal is activated bilaterally, suggesting that PKD1L1 is not required for LPM Nodal pathway activation per se, but rather to restrict Nodal to the left side downstream of nodal flow. Epistasis analysis shows that Pkd1l1 acts as an upstream genetic repressor of Pkd2. This study therefore provides a genetic pathway for the early stages of L-R determination. Moreover, using a system in which cultured cells are supplied artificial flow, we demonstrate that PKD1L1 is sufficient to mediate a Ca2+ signaling response after flow stimulation. Finally, we show that an extracellular PKD domain within PKD1L1 is crucial for PKD1L1 function; as such, destabilizing the domain causes L-R defects in the mouse. Our demonstration that PKD1L1 protein can mediate a response to flow coheres with a mechanosensation model of flow sensation in which the force of fluid flow drives asymmetric gene expression in the embryo. PMID:27272319

  6. Global genetic analysis in mice unveils central role for cilia in congenital heart disease

    PubMed Central

    Li, You; Klena, Nikolai T.; Gabriel, George C; Liu, Xiaoqin; Kim, Andrew J.; Lemke, Kristi; Chen, Yu; Chatterjee, Bishwanath; Devine, William; Damerla, Rama Rao; Chang, Chien-fu; Yagi, Hisato; San Agustin, Jovenal T.; Thahir, Mohamed; Anderton, Shane; Lawhead, Caroline; Vescovi, Anita; Pratt, Herbert; Morgan, Judy; Haynes, Leslie; Smith, Cynthia L.; Eppig, Janan T.; Reinholdt, Laura; Francis, Richard; Leatherbury, Linda; Ganapathiraju, Madhavi K.; Tobita, Kimimasa; Pazour, Gregory J.; Lo, Cecilia W.

    2015-01-01

    Congenital heart disease (CHD) is the most prevalent birth defect, affecting nearly 1% of live births1, but the incidence of CHD is up to ten fold higher in human fetuses2,3. A genetic contribution is strongly suggested by the association of CHD with chromosome abnormalities and high recurrence risk4. Here we report findings from a recessive forward genetic screen in fetal mice, showing the cilium and cilia transduced cell signaling play important roles in the pathogenesis of CHD. The cilium is an evolutionarily conserved organelle projecting from the cell surface with essential roles in diverse cellular processes. Using echocardiography, we ultrasound scanned 87,355 chemically mutagenized C57BL/6J fetal mice and recovered 218 CHD mouse models. Whole exome sequencing identified 91 recessive CHD mutations in 61 genes. This included 34 cilia-related genes, 16 genes involved in cilia transduced cell signaling, and 10 genes regulating vesicular trafficking, a pathway important for ciliogenesis and cell signaling. Surprisingly, many CHD genes encoded interacting proteins, suggesting an interactome protein network may provide a larger genomic context for CHD pathogenesis. These findings provide novel insights into the potential Mendelian genetic contribution to CHD in the fetal population, a segment of the human population not well studied. We note pathways identified show overlap with CHD candidate genes recovered in CHD patients5, suggesting they may have relevance to the more complex genetics of CHD overall. These CHD mouse models and >8,000 incidental mutations are sperm archived, creating a rich public resource for human disease modeling. PMID:25807483

  7. Macrophage polarisation by fatty acids is PPARgamma-dependent.

    PubMed

    Pararasa, Chatyan; Bailey, Clifford; Griffiths, Helen

    2014-10-01

    Elevated plasma free fatty acids (FAs) are associated with increased risk of cardiovascular disease. We investigated the effects of the saturated FA palmitate and unsaturated FA oleate on monocyte phenotype and function. Palmitate increased cell surface expression of integrin CD11b and scavenger receptor CD36 in a concentration-dependent manner with some decrease in mitochondrial reducing capacity at high concentration (300µM). Monocytes incubated with palmitate, but not oleate, showed increased uptake of oxidized LDL and increased adhesion to rat aortic endothelium, particularly at bifurcations. The palmitate-induced increase in CD11b and CD36 expression was associated with increased cellular C16 ceramide and sphingomyelin, loss of reduced glutathione, and increased reactive oxygen species (ROS). Increased monocyte surface CD11b and CD36 was inhibited by fumonisin B1, an inhibitor of de novo ceramide synthesis, but not by the superoxide dismutase mimetic MnTBap. In contrast, MnTBap prevented the mitochondrial ROS increase and metabolic inhibition due to 300µM palmitate. This study demonstrates that in viable monocytes, palmitate but not oleate increases expression of surface CD11b and CD36. Palmitate increases monocyte adhesion to the aortic wall and promotes uptake of oxidized LDL and this involves de novo ceramide synthesis. We have also explored whether specific dietary fatty acids drive monocyte to macrophage polarisation via metabolic pathways. Here we show that monocytes pre-incubated with the saturated fatty acid palmitate increase production of inflammatory cytokines such as TNFa and IL-6 in response to a phorbol myristate differentiation trigger. This increases mitochondrial superoxide production, reduces dependency on oxidative phosphorylation through ceramide-dependent inhibition of PPARgamma activity and increases TNFa production, again via a mechanism that requires ceramide production. PMID:26461339

  8. The roles of evolutionarily conserved functional modules in cilia-related trafficking

    PubMed Central

    Sung, Ching-Hwa; Leroux, Michel R.

    2014-01-01

    Cilia are present across most eukaryotic phyla and have diverse sensory and motility roles in animal physiology, cell signalling and development. Their biogenesis and maintenance depend on vesicular and intraciliary (intraflagellar) trafficking pathways that share conserved structural and functional modules. The functional units of the interconnected pathways, which include proteins involved in membrane coating as well as small GTPases and their accessory factors, were first experimentally associated with canonical vesicular trafficking. These components are, however, ancient, having been co-opted by the ancestral eukaryote to establish the ciliary organelle, and their study can inform us about ciliary biology in higher organisms. PMID:24296415

  9. Robust and stretchable indium gallium zinc oxide-based electronic textiles formed by cilia-assisted transfer printing.

    PubMed

    Yoon, Jongwon; Jeong, Yunkyung; Kim, Heeje; Yoo, Seonggwang; Jung, Hoon Sun; Kim, Yonghun; Hwang, Youngkyu; Hyun, Yujun; Hong, Woong-Ki; Lee, Byoung Hun; Choa, Sung-Hoon; Ko, Heung Cho

    2016-01-01

    Electronic textile (e-textile) allows for high-end wearable electronic devices that provide easy access for carrying, handling and using. However, the related technology does not seem to be mature because the woven fabric hampers not only the device fabrication process directly on the complex surface but also the transfer printing of ultrathin planar electronic devices. Here we report an indirect method that enables conformal wrapping of surface with arbitrary yet complex shapes. Artificial cilia are introduced in the periphery of electronic devices as adhesive elements. The cilia also play an important role in confining a small amount of glue and damping mechanical stress to maintain robust electronic performance under mechanical deformation. The example of electronic applications depicts the feasibility of cilia for 'stick-&-play' systems, which provide electronic functions by transfer printing on unconventional complex surfaces. PMID:27248982

  10. Robust and stretchable indium gallium zinc oxide-based electronic textiles formed by cilia-assisted transfer printing

    PubMed Central

    Yoon, Jongwon; Jeong, Yunkyung; Kim, Heeje; Yoo, Seonggwang; Jung, Hoon Sun; Kim, Yonghun; Hwang, Youngkyu; Hyun, Yujun; Hong, Woong-Ki; Lee, Byoung Hun; Choa, Sung-Hoon; Ko, Heung Cho

    2016-01-01

    Electronic textile (e-textile) allows for high-end wearable electronic devices that provide easy access for carrying, handling and using. However, the related technology does not seem to be mature because the woven fabric hampers not only the device fabrication process directly on the complex surface but also the transfer printing of ultrathin planar electronic devices. Here we report an indirect method that enables conformal wrapping of surface with arbitrary yet complex shapes. Artificial cilia are introduced in the periphery of electronic devices as adhesive elements. The cilia also play an important role in confining a small amount of glue and damping mechanical stress to maintain robust electronic performance under mechanical deformation. The example of electronic applications depicts the feasibility of cilia for ‘stick-&-play' systems, which provide electronic functions by transfer printing on unconventional complex surfaces. PMID:27248982

  11. Robust and stretchable indium gallium zinc oxide-based electronic textiles formed by cilia-assisted transfer printing

    NASA Astrophysics Data System (ADS)

    Yoon, Jongwon; Jeong, Yunkyung; Kim, Heeje; Yoo, Seonggwang; Jung, Hoon Sun; Kim, Yonghun; Hwang, Youngkyu; Hyun, Yujun; Hong, Woong-Ki; Lee, Byoung Hun; Choa, Sung-Hoon; Ko, Heung Cho

    2016-06-01

    Electronic textile (e-textile) allows for high-end wearable electronic devices that provide easy access for carrying, handling and using. However, the related technology does not seem to be mature because the woven fabric hampers not only the device fabrication process directly on the complex surface but also the transfer printing of ultrathin planar electronic devices. Here we report an indirect method that enables conformal wrapping of surface with arbitrary yet complex shapes. Artificial cilia are introduced in the periphery of electronic devices as adhesive elements. The cilia also play an important role in confining a small amount of glue and damping mechanical stress to maintain robust electronic performance under mechanical deformation. The example of electronic applications depicts the feasibility of cilia for `stick-&-play' systems, which provide electronic functions by transfer printing on unconventional complex surfaces.

  12. Localization of a Guanylyl Cyclase to Chemosensory Cilia Requires the Novel Ciliary MYND Domain Protein DAF-25

    PubMed Central

    Jensen, Victor L.; Bialas, Nathan J.; Bishop-Hurley, Sharon L.; Molday, Laurie L.; Kida, Katarzyna; Nguyen, Phuong Anh T.; Blacque, Oliver E.; Molday, Robert S.; Leroux, Michel R.; Riddle, Donald L.

    2010-01-01

    In harsh conditions, Caenorhabditis elegans arrests development to enter a non-aging, resistant diapause state called the dauer larva. Olfactory sensation modulates the TGF-β and insulin signaling pathways to control this developmental decision. Four mutant alleles of daf-25 (abnormal DAuer Formation) were isolated from screens for mutants exhibiting constitutive dauer formation and found to be defective in olfaction. The daf-25 dauer phenotype is suppressed by daf-10/IFT122 mutations (which disrupt ciliogenesis), but not by daf-6/PTCHD3 mutations (which prevent environmental exposure of sensory cilia), implying that DAF-25 functions in the cilia themselves. daf-25 encodes the C. elegans ortholog of mammalian Ankmy2, a MYND domain protein of unknown function. Disruption of DAF-25, which localizes to sensory cilia, produces no apparent cilia structure anomalies, as determined by light and electron microscopy. Hinting at its potential function, the dauer phenotype, epistatic order, and expression profile of daf-25 are similar to daf-11, which encodes a cilium-localized guanylyl cyclase. Indeed, we demonstrate that DAF-25 is required for proper DAF-11 ciliary localization. Furthermore, the functional interaction is evolutionarily conserved, as mouse Ankmy2 interacts with guanylyl cyclase GC1 from ciliary photoreceptors. The interaction may be specific because daf-25 mutants have normally-localized OSM-9/TRPV4, TAX-4/CNGA1, CHE-2/IFT80, CHE-11/IFT140, CHE-13/IFT57, BBS-8, OSM-5/IFT88, and XBX-1/D2LIC in the cilia. Intraflagellar transport (IFT) (required to build cilia) is not defective in daf-25 mutants, although the ciliary localization of DAF-25 itself is influenced in che-11 mutants, which are defective in retrograde IFT. In summary, we have discovered a novel ciliary protein that plays an important role in cGMP signaling by localizing a guanylyl cyclase to the sensory organelle. PMID:21124868

  13. Mapping gravitational-wave backgrounds of arbitrary polarisation using pulsar timing arrays

    NASA Astrophysics Data System (ADS)

    Gair, Jonathan R.; Romano, Joseph D.; Taylor, Stephen R.

    2015-11-01

    We extend our previous work on mapping gravitational-wave backgrounds using techniques borrowed from the analysis of cosmic microwave background data to backgrounds which have non-general-relativity (non-GR) polarisations. Our analysis and results are presented in the context of pulsar timing array observations, but the overarching methods are general, and can be easily applied to LIGO or eLISA observations using appropriately modified response functions. Analytic expressions for the pulsar timing response to gravitational waves with non-GR polarisation are given for each mode of a spin-weighted spherical-harmonic decomposition of the background, which permit the signal to be mapped across the sky to any desired resolution. We also derive the pulsar timing overlap reduction functions for the various non-GR polarisations, finding analytic forms for anisotropic backgrounds with scalar-transverse ("breathing") and vector-longitudinal polarisations, and a semianalytic form for scalar-longitudinal backgrounds. Our results indicate that pulsar timing observations will be completely insensitive to scalar-transverse mode anisotropies in the polarisation amplitude beyond dipole, and anisotropies in the power beyond quadrupole. Analogous to our previous findings that pulsar timing observations lack sensitivity to tensor-curl modes for a transverse-traceless tensor background, we also find insensitivity to vector-curl modes for a vector-longitudinal background.

  14. Circularly polarised printed antenna with wide axial-ratio bandwidth using strip dipoles and slots

    NASA Astrophysics Data System (ADS)

    Ito, K.

    1983-10-01

    A circularly polarised series-fed printed-array antenna having a wide axial-ratio bandwidth (for an axial ratio of less than 3 dB) is realised by combining strip dipoles with slots. To form a free-space dipole, i.e. a complementary radiator to a slot, the strip dipole is constructed by modifying the strip radiator of a circularly polarised antenna. Several experiments verify that the strip dipole behaves like a free-space dipole. A design procedure of the circularly polarised printed antenna with a wide axial-ratio bandwidth is described. On the basis of the design procedure, two types of the array antenna having different strip-dipole spacings are made and measured at S-band. An axial-ratio bandwidth of about 27 percent is obtained for the three sets of the radiators, although the actual gain is less than 0 dBi because the impedance matching is neglected.

  15. Effects of frequency, irradiation geometry and polarisation on computation of SAR in human brain.

    PubMed

    Zhou, Hongmei; Su, Zhentao; Ning, Jing; Wang, Changzhen; Xie, Xiangdong; Qu, Decheng; Wu, Ke; Zhang, Xiaomin; Pan, Jie; Yang, Guoshan

    2014-12-01

    The power absorbed by the human brain has possible implications in the study of the central nervous system-related biological effects of electromagnetic fields. In order to determine the specific absorption rate (SAR) of radio frequency (RF) waves in the human brain, and to investigate the effects of geometry and polarisation on SAR value, the finite-difference time-domain method was applied for the SAR computation. An anatomically realistic model scaled to a height of 1.70 m and a mass of 63 kg was selected, which included 14 million voxels segmented into 39 tissue types. The results suggested that high SAR values were found in the brain, i.e. ∼250 MHz for vertical polarisation and 900-1200 MHz both for vertical and horizontal polarisation, which may be the result of head resonance at these frequencies. PMID:24399107

  16. Efficient, polarised, gain-switched operation of a Tm-doped fibre laser.

    PubMed

    Simakov, Nikita; Hemming, Alex; Bennetts, Shayne; Haub, John

    2011-08-01

    We present a monolithic, robustly polarised thulium fibre laser which produces pulses with 25 ns duration and energy of up to 35 µJ. A pulsed 1.55 μm source was used to gain switch the laser at repetition rates of up to 300 kHz, producing average powers of up to 8 W at 2.044 µm. To the best of our knowledge this is the highest power gain-switched thulium fibre laser and it is also the first with a linearly polarised output. The large repetition rate, pulse energy and average power will enable efficient energy extraction from a large mode-area, polarisation maintaining thulium fibre amplifier. PMID:21934856

  17. Structure of hydrogenous liquids: separation of coherent and incoherent cross sections using polarised neutrons

    NASA Astrophysics Data System (ADS)

    Stunault, A.; Vial, S.; Pusztai, L.; Cuello, G. J.; Temleitner, L.

    2016-04-01

    The determination of the coherent structure factor of hydrogenous liquids is very difficult: while X-rays are barely sensitive to hydrogen, neutrons results still lack accuracy due to the contamination of the scattering intensities by a huge spin-incoherent signal from the 1H atoms. Using polarised neutrons with polarisation analysis, one can experimentally separate the coherent and incoherent contributions to the scattered intensity. We present the upgrade of the D3 polarised hot neutron diffractometer at ILL to study hydrogenated liquids. We show first data obtained from a test sample of water and detail the data reduction leading to an unprecedented accuracy in the extraction of the coherent signal, representative of the structure.

  18. Fabrication-tolerant integrated polarisation splitter based on cascaded Mach–Zehnder interferometers

    SciTech Connect

    Koshelev, A Yu; Gol'tsov, A Yu

    2013-12-31

    We report a fabrication-tolerant polarisation splitter based on cascaded Mach – Zehnder interferometers. This configuration enables a factor of 2 – 3 increase (at the 20 dB level) in the tolerance to the phase difference in comparison with a single interferometer. As an example, we present numerical simulation of a splitter with a centre wavelength of 650 nm, based on a planar waveguide from Si{sub 3}N{sub 4}. The permissible channel waveguide width deviation from calculation results (20-dB extinction coefficient bandwidth) is ∼8% (∼30 nm) for the TE polarisation and ∼30% (100 nm) for the TM polarisation. (fiber and integrated optics)

  19. Modelling the circular polarisation of Earth-like exoplanets: constraints on detecting homochirality

    NASA Astrophysics Data System (ADS)

    Hogenboom, Michael; Stam, Daphne; Rossi, Loic; Snik, Frans

    2016-04-01

    The circular polarisation of light is a property of electromagnetic radiation from which extensive information can be extracted. It is oft-neglected due to its small signal relative to linear polarisation and the need for advanced instrumentation in measuring it. Additionally, numerical modelling is complex as the full Stokes vector must always be computed. Circular polarisation is commonly induced through the multiple scattering of light by aerosols te{hansen} and multiple reflections of light by rough surfaces te{circplanets}. Most interestingly, distinctive spectral circular polarimetric behaviour is exhibited by light reflected by organisms due to the homochiral molecular structure of all known organisms te{chiralbailey}. Especially fascinating is the unique circular polarimetric behaviour of light reflected by photosynthesising organisms at the absorption wavelength of the chlorophyll pigment te{circpolchar}. This presents the previously unexplored possibility of circular polarimetry as a method for identifying and characterising the presence of organisms, a method which could be applied in the hunt for extraterrestrial life. To date, few telescopes exist that measure circular polarisation and none that have been deployed in space. Observations of the circular polarisation reflected by other planets in the solar system have been made with ground-based telescopes, with significant results te{circplanets}. However, none of these observations have been made at the phase angles at which exoplanets will be observed. Also, none have been made of the Earth, which is the logical starting point for the study of biologically induced circular polarisation signals. This introduces the need for numerical modelling to determine the extent to which circular polarisation is present in light reflected by exoplanets or the Earth. In this study, we model the multiple scattering and reflection of light using the doubling-adding method te{dehaan}. We will present circular

  20. Lipopolysaccharide (LPS) disrupts particle transport, cilia function and sperm motility in an ex vivo oviduct model

    PubMed Central

    O’Doherty, A. M.; Di Fenza, M.; Kölle, S.

    2016-01-01

    The oviduct functions in the transportation of gametes to the site of fertilization (the ampulla) and is the site of early embryonic development. Alterations of this early developmental environment, such as the presence of sexually transmitted pathogens, may affect oviduct function leading to reduced fertilization rates and contribute to compromised embryonic development. In this study, sperm interactions, particle transport speed (PTS) and cilia beat frequency (CBF) in the ampulla following exposure to lipopolysaccharide (LPS), a constituent of the sexually transmitted pathogens Chlamydia trachomatis and Chlamydia abortus, was investigated. Three complementary experiments were performed to analyse; (1) bound sperm motility and cilia function (2) transport velocity in the oviduct and (3) the expression of genes related to immune function and inflammatory response (CASP3, CD14, MYD88, TLR4 and TRAF6). The motility of bound sperm was significantly lower in ampullae that were exposed to LPS. CBF and PTS significantly increased after treatment with LPS for 2 hours. Finally, gene expression analysis revealed that CASP3 and CD14 were significantly upregulated and TLR4 trended towards increased expression following treatment with LPS. These findings provide an insight on the impact of LPS on the oviduct sperm interaction, and have implications for both male and female fertility. PMID:27079521

  1. Co-expression of anoctamins in cilia of olfactory sensory neurons.

    PubMed

    Henkel, Bastian; Drose, Daniela R; Ackels, Tobias; Oberland, Sonja; Spehr, Marc; Neuhaus, Eva M

    2015-02-01

    Vertebrates can sense and identify a vast array of chemical cues. The molecular machinery involved in chemodetection and transduction is expressed within the cilia of olfactory sensory neurons. Currently, there is only limited information available on the distribution and density of individual signaling components within the ciliary compartment. Using super-resolution microscopy, we show here that cyclic-nucleotide-gated channels and calcium-activated chloride channels of the anoctamin family are localized to discrete microdomains in the ciliary membrane. In addition to ANO2, a second anoctamin, ANO6, also localizes to ciliary microdomains. This observation, together with the fact that ANO6 and ANO2 co-localize, indicates a role for ANO6 in olfactory signaling. We show that both ANO2 and ANO6 can form heteromultimers and that this heteromerization alters the recombinant channels' physiological properties. Thus, we provide evidence for interaction of ANO2 and ANO6 in olfactory cilia, with possible physiological relevance for olfactory signaling. PMID:25500808

  2. The effect of cilia and the mucociliary clearance on successful drug delivery.

    PubMed

    Gizurarson, Sveinbjörn

    2015-01-01

    Nasal mucociliary clearance is one of the most important factors affecting nasal delivery of drugs and vaccines. This is also the most important physiological defense mechanism inside the nasal cavity. It removes inhaled (and delivered) particles, microbes and substances trapped in the mucus. Almost all inhaled particles are trapped in the mucus carpet and transported with a rate of 8-10 mm/h toward the pharynx. This transport is conducted by the ciliated cells, which contain about 100-250 motile cellular appendages called cilia, 0.3 µm wide and 5 µm in length that beat about 1000 times every minute or 12-15 Hz. For efficient mucociliary clearance, the interaction between the cilia and the nasal mucus needs to be well structured, where the mucus layer is a tri-layer: an upper gel layer that floats on the lower, more aqueous solution, called the periciliary liquid layer and a third layer of surfactants between these two main layers. Pharmacokinetic calculations of the mucociliary clearance show that this mechanism may account for a substantial difference in bioavailability following nasal delivery. If the formulation irritates the nasal mucosa, this mechanism will cause the irritant to be rapidly diluted, followed by increased clearance, and swallowed. The result is a much shorter duration inside the nasal cavity and therefore less nasal bioavailability. PMID:25739664

  3. Cilia play a role in breaking left-right symmetry of the sea urchin embryo.

    PubMed

    Takemoto, Ayumi; Miyamoto, Tatsuo; Simono, Fumie; Kurogi, Nao; Shirae-Kurabayashi, Maki; Awazu, Akinori; Suzuki, Ken-Ichi T; Yamamoto, Takashi; Sakamoto, Naoaki

    2016-06-01

    Left-right asymmetry of bilaterian animals is established during early development. In mice, frogs and fishes, the ciliated left-right organizer plays an essential role in establishing bilateral asymmetry, and leftward flow of extracellular fluid generated by ciliary motion results in Nodal activity on the left side. However, H(+) /K(+) -ATPase activity is also involved in the determination of left-right asymmetry in a variety of animals, and it has been thought to be an ancestral mechanism in deuterostomes. In sea urchin, the determination of the left-right asymmetry based on H(+) /K(+) -ATPase activity was already clarified, but it remains to be uncovered whether ciliary motion is involved in the left-right asymmetry of the embryo. Here, we show evidence that ciliary motion is involved in the establishment of left-right asymmetry of sea urchin embryo. Furthermore, we show that the initial cilia generated on small micromeres during the early stage of embryogenesis may be involved in this process. These results suggest that the cilia-mediated mechanism for the determination of left-right asymmetry may be acquired at the base of the deuterostomes. PMID:27028068

  4. Early eukaryotic origins for cilia-associated bioactive peptide-amidating activity.

    PubMed

    Kumar, Dhivya; Blaby-Haas, Crysten E; Merchant, Sabeeha S; Mains, Richard E; King, Stephen M; Eipper, Betty A

    2016-03-01

    Ciliary axonemes and basal bodies were present in the last eukaryotic common ancestor and play crucial roles in sensing and responding to environmental cues. Peptidergic signaling, generally considered a metazoan innovation, is essential for organismal development and homeostasis. Peptidylglycine α-amidating monooxygenase (PAM) is crucial for the last step of bioactive peptide biosynthesis. However, identification of a complete PAM-like gene in green algal genomes suggests ancient evolutionary roots for bioactive peptide signaling. We demonstrate that the Chlamydomonas reinhardtii PAM gene encodes an active peptide-amidating enzyme (CrPAM) that shares key structural and functional features with the mammalian enzyme, indicating that components of the peptide biosynthetic pathway predate multicellularity. In addition to its secretory pathway localization, CrPAM localizes to cilia and tightly associates with the axonemal superstructure, revealing a new axonemal enzyme activity. This localization pattern is conserved in mammals, with PAM present in both motile and immotile sensory cilia. The conserved ciliary localization of PAM adds to the known signaling capabilities of the eukaryotic cilium and provides a potential mechanistic link between peptidergic signaling and endocrine abnormalities commonly observed in ciliopathies. PMID:26787743

  5. The Tumor Suppressor pVHL Down-regulates Never-in-Mitosis A-related Kinase 8 via Hypoxia-inducible Factors to Maintain Cilia in Human Renal Cancer Cells*

    PubMed Central

    Ding, Xiao-Fei; Zhou, Jun; Hu, Qiong-Ying; Liu, Shuang-Chun; Chen, Guang

    2015-01-01

    NEK8 (never in mitosis gene A (NIMA)-related kinase 8) is involved in cytoskeleton, cilia, and DNA damage response/repair. Abnormal expression and/or dysfunction of NEK8 are related to cancer development and progression. However, the mechanisms that regulate NEK8 are not well declared. We demonstrated here that pVHL may be involved in regulating NEK8. We found that CAK-I cells with wild-type vhl expressed a lower level of NEK8 than the cells loss of vhl, such as 786-O, 769-P, and A-498 cells. Moreover, pVHL overexpression down-regulated the NEK8 protein in 786-O cells, whereas pVHL knockdown up-regulated NEK8 in CAK-I cells. In addition, we found that the positive hypoxia response elements (HREs) are located in the promoter of the nek8 sequence and hypoxia could induce nek8 expression in different cell types. Consistent with this, down-regulation of hypoxia-inducible factors α (HIF-1α or HIF-2α) by isoform-specific siRNA reduced the ability of hypoxia inducing nek8 expression. In vivo, NEK8 and HIF-1α expression were increased in kidneys of rats subjected to an experimental hypoxia model of ischemia and reperfusion. Furthermore, NEK8 siRNA transfection significantly blocked pVHL-knockdown-induced cilia disassembling, through impairing the pVHL-knockdown-up-regulated NEK8 expression. These results support that nek8 may be a novel hypoxia-inducible gene. In conclusion, our findings show that nek8 may be a new HIF target gene and pVHL can down-regulate NEK8 via HIFs to maintain the primary cilia structure in human renal cancer cells. PMID:25451921

  6. The tumor suppressor pVHL down-regulates never-in-mitosis A-related kinase 8 via hypoxia-inducible factors to maintain cilia in human renal cancer cells.

    PubMed

    Ding, Xiao-Fei; Zhou, Jun; Hu, Qiong-Ying; Liu, Shuang-Chun; Chen, Guang

    2015-01-16

    NEK8 (never in mitosis gene A (NIMA)-related kinase 8) is involved in cytoskeleton, cilia, and DNA damage response/repair. Abnormal expression and/or dysfunction of NEK8 are related to cancer development and progression. However, the mechanisms that regulate NEK8 are not well declared. We demonstrated here that pVHL may be involved in regulating NEK8. We found that CAK-I cells with wild-type vhl expressed a lower level of NEK8 than the cells loss of vhl, such as 786-O, 769-P, and A-498 cells. Moreover, pVHL overexpression down-regulated the NEK8 protein in 786-O cells, whereas pVHL knockdown up-regulated NEK8 in CAK-I cells. In addition, we found that the positive hypoxia response elements (HREs) are located in the promoter of the nek8 sequence and hypoxia could induce nek8 expression in different cell types. Consistent with this, down-regulation of hypoxia-inducible factors α (HIF-1α or HIF-2α) by isoform-specific siRNA reduced the ability of hypoxia inducing nek8 expression. In vivo, NEK8 and HIF-1α expression were increased in kidneys of rats subjected to an experimental hypoxia model of ischemia and reperfusion. Furthermore, NEK8 siRNA transfection significantly blocked pVHL-knockdown-induced cilia disassembling, through impairing the pVHL-knockdown-up-regulated NEK8 expression. These results support that nek8 may be a novel hypoxia-inducible gene. In conclusion, our findings show that nek8 may be a new HIF target gene and pVHL can down-regulate NEK8 via HIFs to maintain the primary cilia structure in human renal cancer cells. PMID:25451921

  7. Polarisation of the auroral red line in the Earth's upper atmosphere: a review (Invited)

    NASA Astrophysics Data System (ADS)

    Lamy, H.; Barthelemy, M.; Lilensten, J.; Bommier, V.; Simon Wedlund, C.

    2013-12-01

    Polarisation of light is a key observable to provide information about asymmetry or anisotropy within a radiative source. Polarimetry of auroral emission lines in the Earth's upper atmosphere has been overlooked for decades. However, the bright red auroral line (6300Å) produced by collisional impact with electrons precipitating along magnetic field lines is a good candidate to search for polarisation. This problem was investigated recently with observations obtained by Lilensten et al (2008), Barthélemy et al (2011) and Lilensten et al (2013) with a photopolarimeter. Analysis of the data indicates that the red auroral emission line is polarised at a level of a few percent. The results are compared to theoretical predictions of Bommier et al (2011) that were obtained for a collimated beam. The comparison suggests the existence of depolarization processes whose origin will be discussed. A new dedicated spectropolarimeter currently under development will also be presented. This instrument will cover the optical spectrum from approximately 400 to 700 nm providing simultaneously the polarisation of the red line and of other interesting auroral emission lines such as N2+ 1NG (4278Å), other N2 bands, etc... The importance of these polarisation measurements in the context of upper atmosphere modelling and geomagnetic activity will be discussed. Lilensten, J. et al, Polarization in aurorae: A new dimension for space environments studies, Geophys. Res. Lett., 26, 269, 2008 Barthélemy M. et al, Polarisation in the auroral red line during coordinated EISCAT Svalbard Radar/optical experiments, Annales Geophysicae, Volume 29, Issue 6, 2011, 1101-1112, 2011. Bommier V. et al, The Theoretical Impact Polarization of the O I 6300 Å Red Line of Earth Auroræ, Annales Geophysicae, Volume 29, Issue 1, 2011, 71-79, 2011 Lilensten, J. et al, The thermospheric auroral red line polarization: confirmation of detection and first quantitative analysis, Journal of Space Weather and Space

  8. Polarisation splitting of laser beams by large angles with minimal reflection losses

    SciTech Connect

    Davydov, B L

    2006-05-31

    New crystal anisotropic prisms for splitting orthogonally polarised components of laser radiation by large angles with minimal reflection losses caused by the Brewster refraction and total internal reflection of polarised waves from the crystal-air interface are considered and the method for their calculation is described. It is shown that, by assembling glue-free combinations of two or three prisms, thermally stable beamsplitters can be fabricated, which are free from the beam astigmatism and the wave dispersion of the output angles of the beams. The parameters and properties of new beamsplitters are presented in a convenient form in figures and tables. (laser applications and other topics in quantum electronics)

  9. Resonance method to produce a polarisation asymmetry in electron-positron storage rings

    SciTech Connect

    Toner, W.T.

    1988-01-01

    Pulsed solenoids of a few tens of ampere turns, operated in synchronism with the ..gamma..(g-2/2) 'th harmonic of the orbit period, can be used to prevent a stored electron beam from becoming polarised through the emission of synchrotron radiation. With such low fields it is easy to arrange that only some of the stored bunches are affected. This makes it possible to produce collisions between counter-rotating electrons and positrons stored in a single ring in which the electron and positron polarisations are not equal and opposite. 8 refs.

  10. Precise measurement of CMB polarisation from Dome-C: the BRAIN and CLOVER experiments

    NASA Astrophysics Data System (ADS)

    Piat, M.; Rosset, C.; Bartlett, J.; Giraud-Héraud, Y.; Bréele, E.; Maestrini, A.; Tripon Canseliet, C.; Algani, C.; Girard, M.; Pajot, F.; Masi, S.; de Bernardis, P.; Piccirillo, L.; Mafei, B.; Jones, M.; Taylor, A.

    2004-12-01

    The characterisation of CMB polarisation is one of the next challenge in observational cosmology. This is especially true for the so-called B-modes that are at least 3 order of magnitude lower than CMB temperature fluctuations. A precise measurement of the angular power spectrum of these B-modes will give important constraints on inflation parameters. In this talk, I will describe two complementary experiments, BRAIN and CLOVER, dedicated to CMB polarisation measurement. These experiments are proposed to be installed in Dome-C, Antarctica, to take advantage of the extreme dryness of the atmosphere and to allow long integration time.

  11. A prefoldin-associated WD-repeat protein (WDR92) is required for the correct architectural assembly of motile cilia.

    PubMed

    Patel-King, Ramila S; King, Stephen M

    2016-04-15

    WDR92 is a highly conserved WD-repeat protein that has been proposed to be involved in apoptosis and also to be part of a prefoldin-like cochaperone complex. We found that WDR92 has a phylogenetic signature that is generally compatible with it playing a role in the assembly or function of specifically motile cilia. To test this hypothesis, we performed an RNAi-based knockdown of WDR92 gene expression in the planarianSchmidtea mediterraneaand were able to achieve a robust reduction in mRNA expression to levels undetectable under our standard RT-PCR conditions. We found that this treatment resulted in a dramatic reduction in the rate of organismal movement that was caused by a switch in the mode of locomotion from smooth, cilia-driven gliding to muscle-based, peristaltic contractions. Although the knockdown animals still assembled cilia of normal length and in similar numbers to controls, these structures had reduced beat frequency and did not maintain hydrodynamic coupling. By transmission electron microscopy we observed that many cilia had pleiomorphic defects in their architecture, including partial loss of dynein arms, incomplete closure of the B-tubule, and occlusion or replacement of the central pair complex by accumulated electron-dense material. These observations suggest that WDR92 is part of a previously unrecognized cytoplasmic chaperone system that is specifically required to fold key components necessary to build motile ciliary axonemes. PMID:26912790

  12. The PDZ Protein Na+/H+ Exchanger Regulatory Factor-1 (NHERF1) Regulates Planar Cell Polarity and Motile Cilia Organization

    PubMed Central

    Stolz, Donna B.; Tsang, Michael; Friedman, Peter A.; Romero, Guillermo

    2016-01-01

    Directional flow of the cerebrospinal fluid requires coordinated movement of the motile cilia of the ependymal epithelium that lines the cerebral ventricles. Here we report that mice lacking the Na+/H+ Exchanger Regulatory Factor 1 (NHERF1/Slc9a3r1, also known as EBP50) develop profound communicating hydrocephalus associated with fewer and disorganized ependymal cilia. Knockdown of NHERF1/slc9a3r1 in zebrafish embryos also causes severe hydrocephalus of the hindbrain and impaired ciliogenesis in the otic vesicle. Ultrastructural analysis did not reveal defects in the shape or organization of individual cilia. Similar phenotypes have been described in animals with deficiencies in Wnt signaling and the Planar Cell Polarity (PCP) pathway. We show that NHERF1 binds the PCP core genes Frizzled (Fzd) and Vangl. We further show that NHERF1 assembles a ternary complex with Fzd4 and Vangl2 and promotes translocation of Vangl2 to the plasma membrane, in particular to the apical surface of ependymal cells. Taken together, these results strongly support an important role for NHERF1 in the regulation of PCP signaling and the development of functional motile cilia. PMID:27055101

  13. A prefoldin-associated WD-repeat protein (WDR92) is required for the correct architectural assembly of motile cilia

    PubMed Central

    Patel-King, Ramila S.; King, Stephen M.

    2016-01-01

    WDR92 is a highly conserved WD-repeat protein that has been proposed to be involved in apoptosis and also to be part of a prefoldin-like cochaperone complex. We found that WDR92 has a phylogenetic signature that is generally compatible with it playing a role in the assembly or function of specifically motile cilia. To test this hypothesis, we performed an RNAi-based knockdown of WDR92 gene expression in the planarian Schmidtea mediterranea and were able to achieve a robust reduction in mRNA expression to levels undetectable under our standard RT-PCR conditions. We found that this treatment resulted in a dramatic reduction in the rate of organismal movement that was caused by a switch in the mode of locomotion from smooth, cilia-driven gliding to muscle-based, peristaltic contractions. Although the knockdown animals still assembled cilia of normal length and in similar numbers to controls, these structures had reduced beat frequency and did not maintain hydrodynamic coupling. By transmission electron microscopy we observed that many cilia had pleiomorphic defects in their architecture, including partial loss of dynein arms, incomplete closure of the B-tubule, and occlusion or replacement of the central pair complex by accumulated electron-dense material. These observations suggest that WDR92 is part of a previously unrecognized cytoplasmic chaperone system that is specifically required to fold key components necessary to build motile ciliary axonemes. PMID:26912790

  14. A Conditional Mutant Having Paralyzed Cilia and a Block in Cytokinesis Is Rescued by Cytoplasmic Exchange in Tetrahymena Thermophila

    PubMed Central

    Pennock, D. G.; Thatcher, T.; Bowen, J.; Bruns, P. J.; Gorovsky, M. A.

    1988-01-01

    Nineteen mutants that are conditional for both the ability to regain motility following deciliation and the ability to grow were isolated. The mutations causing slow growth were placed into five complementation groups. None of the mutations appears to affect energy production as all mutants remained motile at the restrictive temperature. In three complementation groups protein synthesis and the levels of mRNA encoding α-tubulin or actin were largely unaffected at the restrictive temperature, consistent with the hypothesis that mutations in these three groups directly affect the assembly of functional cilia and growth. Complementation group 1 was chosen for further characterization. Both phenotypes were shown to be linked, suggesting they are caused by a single mutation. Group 1 mutants regenerated cilia at the restrictive temperature, but the cilia were nonmotile. This mutation also caused a block in cytokinesis at the restrictive temperature but did not affect nuclear divisions or DNA synthesis. The block in cell division was transiently rescued by wild-type cytoplasm exchanged when mutants were paired with wild-type cells during conjugation (round 1 of genomic exclusion). Thus, at least one mutation has been isolated that affects assembly of some microtubule-based structures in Tetrahymena (cilia during regeneration) but not others (nuclei divide at 38°), and the product of this gene is likely to play a role in both ciliary function and in cytokinesis. PMID:3224807

  15. Gli2 and Gli3 Localize to Cilia and Require the Intraflagellar Transport Protein Polaris for Processing and Function

    SciTech Connect

    Michaud III, Edward J; Haycraft, Courtney J; Aydin Son, Yesim; Zhang, Qihong; Yoder, Bradley

    2005-01-01

    Intraflagellar transport (IFT) proteins are essential for cilia assembly and have recently been associated with a number of developmental processes, such as left-right axis specification and limb and neural tube patterning. Genetic studies indicate that IFT proteins are required for Sonic hedgehog (Shh)signaling downstream of the Smoothened and Patched membrane proteins but upstream of the Glioma (Gli) transcription factors. However, the role that IFT proteins play in transduction of Shh signaling and the importance of cilia in this process remain unknown. Here we provide insights into the mechanism by which defects in an IFT protein, Tg737/Polaris, affect Shh signaling in the murine limb bud. Our data show that loss of Tg737 results in altered Gli3 processing that abrogates Gli3-mediated repression of Gli1 transcriptional activity. In contrast to the conclusions drawn from genetic analysis, the activity of Gli1 and truncated forms of Gli3 (Gli3R) are unaffected in Tg737 mutants at the molecular level, indicating that Tg737/Polaris is differentially involved in specific activities of the Gli proteins. Most important, a negative regulator of Shh signaling, Suppressor of fused, and the three full-length Gli transcription factors localize to the distal tip of cilia in addition to the nucleus. Thus, our data support a model where cilia have a direct role in Gli processing and Shh signal transduction.

  16. RSPH3 Mutations Cause Primary Ciliary Dyskinesia with Central-Complex Defects and a Near Absence of Radial Spokes

    PubMed Central

    Jeanson, Ludovic; Copin, Bruno; Papon, Jean-François; Dastot-Le Moal, Florence; Duquesnoy, Philippe; Montantin, Guy; Cadranel, Jacques; Corvol, Harriet; Coste, André; Désir, Julie; Souayah, Anissa; Kott, Esther; Collot, Nathalie; Tissier, Sylvie; Louis, Bruno; Tamalet, Aline; de Blic, Jacques; Clement, Annick; Escudier, Estelle; Amselem, Serge; Legendre, Marie

    2015-01-01

    Primary ciliary dyskinesia (PCD) is a rare autosomal-recessive condition resulting from structural and/or functional defects of the axoneme in motile cilia and sperm flagella. The great majority of mutations identified so far involve genes whose defects result in dynein-arm anomalies. By contrast, PCD due to CC/RS defects (those in the central complex [CC] and radial spokes [RSs]), which might be difficult to diagnose, remains mostly unexplained. We identified non-ambiguous RSPH3 mutations in 5 of 48 independent families affected by CC/RS defects. RSPH3, whose ortholog in the flagellated alga Chlamydomonas reinhardtii encodes a RS-stalk protein, is mainly expressed in respiratory and testicular cells. Its protein product, which localizes within the cilia of respiratory epithelial cells, was undetectable in airway cells from an individual with RSPH3 mutations and in whom RSPH23 (a RS-neck protein) and RSPH1 and RSPH4A (RS-head proteins) were found to be still present within cilia. In the case of RSPH3 mutations, high-speed-videomicroscopy analyses revealed the coexistence of immotile cilia and motile cilia with movements of reduced amplitude. A striking feature of the ultrastructural phenotype associated with RSPH3 mutations is the near absence of detectable RSs in all cilia in combination with a variable proportion of cilia with CC defects. Overall, this study shows that RSPH3 mutations contribute to disease in more than 10% of PCD-affected individuals with CC/RS defects, thereby allowing an accurate diagnosis to be made in such cases. It also unveils the key role of RSPH3 in the proper building of RSs and the CC in humans. PMID:26073779

  17. Handedness and situs inversus in primary ciliary dyskinesia.

    PubMed

    McManus, I C; Martin, N; Stubbings, G F; Chung, E M K; Mitchison, H M

    2004-12-22

    ... The limbs on the right side are stronger. [The] cause may be ... [that] ... motion, and abilities of moving, are somewhat holpen from the liver, which lieth on the right side. (Sir Francis Bacon, Sylva sylvarum (1627).)Fifty per cent of people with primary ciliary dyskinesia (PCD) (also known as immotile cilia syndrome or Siewert-Kartagener syndrome) have situs inversus, which is thought to result from absent nodal ciliary rotation and failure of normal symmetry breaking. In a study of 88 people with PCD, only 15.2% of 46 individuals with situs inversus, and 14.3% of 42 individuals with situs solitus, were left handed. Because cerebral lateralization is therefore still present, the nodal cilia cannot be the primary mechanism responsible for symmetry breaking in the vertebrate body. Intriguingly, one behavioural lateralization, wearing a wrist-watch on the right wrist, did correlate with situs inversus. PMID:15615683

  18. Mutation of the MAP kinase DYF-5 affects docking and undocking of kinesin-2 motors and reduces their speed in the cilia of Caenorhabditis elegans.

    PubMed

    Burghoorn, Jan; Dekkers, Martijn P J; Rademakers, Suzanne; de Jong, Ton; Willemsen, Rob; Jansen, Gert

    2007-04-24

    In the cilia of the nematode Caenorhabditis elegans, anterograde intraflagellar transport (IFT) is mediated by two kinesin-2 complexes, kinesin II and OSM-3 kinesin. These complexes function together in the cilia middle segments, whereas OSM-3 alone mediates transport in the distal segments. Not much is known about the mechanisms that compartmentalize the kinesin-2 complexes or how transport by both kinesins is coordinated. Here, we identify DYF-5, a conserved MAP kinase that plays a role in these processes. Fluorescence microscopy and EM revealed that the cilia of dyf-5 loss-of-function (lf) animals are elongated and are not properly aligned into the amphid channel. Some cilia do enter the amphid channel, but the distal ends of these cilia show accumulation of proteins. Consistent with these observations, we found that six IFT proteins accumulate in the cilia of dyf-5(lf) mutants. In addition, using genetic analyses and live imaging to measure the motility of IFT proteins, we show that dyf-5 is required to restrict kinesin II to the cilia middle segments. Finally, we show that, in dyf-5(lf) mutants, OSM-3 moves at a reduced speed and is not attached to IFT particles. We propose that DYF-5 plays a role in the undocking of kinesin II from IFT particles and in the docking of OSM-3 onto IFT particles. PMID:17420466

  19. Polarisation Analysis Neutron Spectrometer, POLANO, at J-PARC - Concept and Magnetic Field Optimisation

    NASA Astrophysics Data System (ADS)

    Ohoyama, K.; Yokoo, T.; Itoh, S.; Nanbu, M.; Iwasa, K.; Ohkawara, M.; Kaneko, N.; Ino, T.; Hayashida, H.; Oku, T.; Kira, H.; Tasaki, S.; Takeda, M.; Kimura, H.; Sato, T. J.

    2016-04-01

    The status of the polarised neutron spectrometer constructed at the Japan Proton Accelerator Research Complex through a collaboration between Tohoku University and KEK will be reported. In particular, the optimisation of magnetic fields to minimise neutron- beam depolarisation using the finite element method will be discussed on the basis of several simulations using the finite element method.

  20. Discrimination of water, ice and aerosols by light polarisation in the CLOUD experiment

    NASA Astrophysics Data System (ADS)

    Nichman, L.; Fuchs, C.; Järvinen, E.; Ignatius, K.; Höppel, N. F.; Dias, A.; Heinritzi, M.; Simon, M.; Tröstl, J.; Wagner, A. C.; Wagner, R.; Williamson, C.; Yan, C.; Bianchi, F.; Connolly, P. J.; Dorsey, J. R.; Duplissy, J.; Ehrhart, S.; Frege, C.; Gordon, H.; Hoyle, C. R.; Kristensen, T. B.; Steiner, G.; Donahue, N. M.; Flagan, R.; Gallagher, M. W.; Kirkby, J.; Möhler, O.; Saathoff, H.; Schnaiter, M.; Stratmann, F.; Tomé, A.

    2015-11-01

    Cloud microphysical processes involving the ice phase in tropospheric clouds are among the major uncertainties in cloud formation, weather and General Circulation Models (GCMs). The simultaneous detection of aerosol particles, liquid droplets, and ice crystals, especially in the small cloud-particle size range below 50 μm, remains challenging in mixed phase, often unstable ice-water phase environments. The Cloud Aerosol Spectrometer with Polarisation (CASPOL) is an airborne instrument that has the ability to detect such small cloud particles and measure their effects on the backscatter polarisation state. Here we operate the versatile Cosmics-Leaving-OUtdoor-Droplets (CLOUD) chamber facility at the European Organisation for Nuclear Research (CERN) to produce controlled mixed phase and other clouds by adiabatic expansions in an ultraclean environment, and use the CASPOL to discriminate between different aerosols, water and ice particles. In this paper, optical property measurements of mixed phase clouds and viscous Secondary Organic Aerosol (SOA) are presented. We report observations of significant liquid - viscous SOA particle polarisation transitions under dry conditions using CASPOL. Cluster analysis techniques were subsequently used to classify different types of particles according to their polarisation ratios during phase transition. A classification map is presented for water droplets, organic aerosol (e.g., SOA and oxalic acid), crystalline substances such as ammonium sulphate, and volcanic ash. Finally, we discuss the benefits and limitations of this classification approach for atmospherically relevant concentration and mixtures with respect to the CLOUD 8-9 campaigns and its potential contribution to Tropical Troposphere Layer (TTL) analysis.

  1. Educational Polarisation in Brisbane: Rawls's Least Advantaged and the Myth of Choice

    ERIC Educational Resources Information Center

    Di Bartolo, Lawrence

    2005-01-01

    A number of researchers have reported increasing socio-economic inequality within Australian society. This result has usually been met with public apathy and political indifference. However, the results of this research conducted in Brisbane shows that the increasing social polarisation occurring in Australian society is being reflected in an…

  2. The history of polarisation measurements: their role in studies of magnetic fields

    NASA Astrophysics Data System (ADS)

    Wielebinski, R.

    2015-03-01

    Radio astronomy gave us new methods to study magnetic fields. Synchrotron radiation, the main cause of comic radio waves, is highly linearly polarised with the `E' vector normal to the magnetic field. The Faraday Effect rotates the `E' vector in thermal regions by the magnetic field in the line of sight. Also the radio Zeeman Effect has been observed.

  3. Reconstruction of the primordial power spectrum using temperature and polarisation data from multiple experiments

    SciTech Connect

    Nicholson, Gavin; Contaldi, Carlo R. E-mail: c.contaldi@imperial.ac.uk

    2009-07-01

    We develop a method to reconstruct the primordial power spectrum, P(k), using both temperature and polarisation data from the joint analysis of a number of Cosmic Microwave Background (CMB) observations. The method is an extension of the Richardson-Lucy algorithm, first applied in this context by Shafieloo and Souradeep [1]. We show how the inclusion of polarisation measurements can decrease the uncertainty in the reconstructed power spectrum. In particular, the polarisation data can constrain oscillations in the spectrum more effectively than total intensity only measurements. We apply the estimator to a compilation of current CMB results. The reconstructed spectrum is consistent with the best-fit power spectrum although we find evidence for a 'dip' in the power on scales k ≈ 0.002 Mpc{sup −1}. This feature appears to be associated with the WMAP power in the region 18 ≤ l ≤ 26 which is consistently below best-fit models. We also forecast the reconstruction for a simulated, Planck-like [2] survey including sample variance limited polarisation data.

  4. Reconstruction of the primordial power spectrum using temperature and polarisation data from multiple experiments

    NASA Astrophysics Data System (ADS)

    Nicholson, Gavin; Contaldi, Carlo R.

    2009-07-01

    We develop a method to reconstruct the primordial power spectrum, P(k), using both temperature and polarisation data from the joint analysis of a number of Cosmic Microwave Background (CMB) observations. The method is an extension of the Richardson-Lucy algorithm, first applied in this context by Shafieloo & Souradeep [1]. We show how the inclusion of polarisation measurements can decrease the uncertainty in the reconstructed power spectrum. In particular, the polarisation data can constrain oscillations in the spectrum more effectively than total intensity only measurements. We apply the estimator to a compilation of current CMB results. The reconstructed spectrum is consistent with the best-fit power spectrum although we find evidence for a `dip' in the power on scales k approx 0.002 Mpc-1. This feature appears to be associated with the WMAP power in the region 18 <= ell <= 26 which is consistently below best-fit models. We also forecast the reconstruction for a simulated, Planck-like [2] survey including sample variance limited polarisation data.

  5. Intra-well relaxation process in magnetic fluids subjected to strong polarising fields

    NASA Astrophysics Data System (ADS)

    Marin, C. N.; Fannin, P. C.; Mălăescu, I.; Barvinschi, P.; Ercuţa, A.

    2012-02-01

    We report on the frequency and field dependent complex magnetic susceptibility measurements of a kerosene-based magnetic fluid with iron oxide nanoparticles, stabilized with oleic acid, in the frequency range 0.1-6 GHz and over the polarising field range of 0-168.4 kA/m. By increasing polarising field, H, a subsidiary loss-peak clearly occurs in the vicinity of the ferromagnetic resonance peak, from which it remains distinct even in strong polarising fields of 168.4 kA/m. This is in contrast to other reported cases in which the intra-well relaxation process is manifested only as a shoulder of the resonance peak, which vanishes in polarising fields larger than that of 100 kA/m. The results of the XRD analysis connected to the anisotropy field results confirm that the investigated sample contains particles of magnetite and of the tetragonal phase of maghemite. Taking into account the characteristics of our sample, the theoretical analysis revealed that the intra-well relaxation process of the small particles of the tetragonal phase of maghemite may be responsible for the subsidiary loss peak of the investigated magnetic fluid.

  6. The intrinsic B-mode polarisation of the Cosmic Microwave Background

    SciTech Connect

    Fidler, Christian; Pettinari, Guido W.; Crittenden, Robert; Koyama, Kazuya; Wands, David; Beneke, Martin E-mail: Guido.Pettinari@sussex.ac.uk E-mail: Kazuya.Koyama@port.ac.uk

    2014-07-01

    We estimate the B-polarisation induced in the Cosmic Microwave Background by the non-linear evolution of density perturbations. Using the second-order Boltzmann code SONG, our analysis incorporates, for the first time, all physical effects at recombination. We also include novel contributions from the redshift part of the Boltzmann equation and from the bolometric definition of the temperature in the presence of polarisation. The remaining line-of-sight terms (lensing and time-delay) have previously been studied and must be calculated non-perturbatively. The intrinsic B-mode polarisation is present independent of the initial conditions and might contaminate the signal from primordial gravitational waves. We find this contamination to be comparable to a primordial tensor-to-scalar ratio of r ≅ 10{sup −7} at the angular scale ℓ ≅ 100, where the primordial signal peaks, and r ≅ 5 × 10{sup −5} at ℓ ≅ 700, where the intrinsic signal peaks. Therefore, we conclude that the intrinsic B-polarisation from second-order effects is not likely to contaminate future searches of primordial gravitational waves.

  7. Polarisation inhomogeneities of a ring resonator and nonreciprocity of counterpropagating waves

    SciTech Connect

    Kuryatov, V N; Sokolov, A L

    2002-04-30

    The nonreciprocity of the characteristics of counterpropagating waves in a ring resonator formed by totally reflecting prisms is analysed. The polarisation-inhomogeneous properties of a prism resonator and their effect on the nonreciprocity of the frequencies of counterpropagating waves are studied. (control of laser radiation parameters)

  8. The intrinsic B-mode polarisation of the Cosmic Microwave Background

    NASA Astrophysics Data System (ADS)

    Fidler, Christian; Pettinari, Guido W.; Beneke, Martin; Crittenden, Robert; Koyama, Kazuya; Wands, David

    2014-07-01

    We estimate the B-polarisation induced in the Cosmic Microwave Background by the non-linear evolution of density perturbations. Using the second-order Boltzmann code SONG, our analysis incorporates, for the first time, all physical effects at recombination. We also include novel contributions from the redshift part of the Boltzmann equation and from the bolometric definition of the temperature in the presence of polarisation. The remaining line-of-sight terms (lensing and time-delay) have previously been studied and must be calculated non-perturbatively. The intrinsic B-mode polarisation is present independent of the initial conditions and might contaminate the signal from primordial gravitational waves. We find this contamination to be comparable to a primordial tensor-to-scalar ratio of r simeq 10-7 at the angular scale l simeq 100, where the primordial signal peaks, and r simeq 5 × 10-5 at l simeq 700, where the intrinsic signal peaks. Therefore, we conclude that the intrinsic B-polarisation from second-order effects is not likely to contaminate future searches of primordial gravitational waves.

  9. Mutations in chemosensory cilia cause resistance to paraquat in nematode Caenorhabditis elegans.

    PubMed

    Fujii, Michihiko; Matsumoto, Yuki; Tanaka, Nanae; Miki, Kensuke; Suzuki, Toshikazu; Ishii, Naoaki; Ayusawa, Dai

    2004-05-01

    The relationship between oxidative stress and longevity is a matter of concern in various organisms. We isolated mutants resistant to paraquat from nematode Caenorhabditis elegans. One mutant named mev-4 was long-lived and showed cross-resistance to heat and Dyf phenotype (defective in dye filling). Genetic and sequence analysis revealed that mev-4 had a nonsense mutation on the che-11 gene, homologues of which are involved in formation of cilia and flagella in other organisms. The paraquat resistance was commonly observed in various Dyf mutants and did not depend on the daf-16 gene, whereas the extension of life span did depend on it. Expression of antioxidant enzyme genes seemed normal. These results suggest that chemosensory neurons are a target of oxidative stress and influence longevity dependent on the daf-16 signaling in C. elegans. PMID:14982934

  10. Basal foot MTOC organizes pillar MTs required for coordination of beating cilia

    PubMed Central

    Clare, Daniel K.; Magescas, Jérémy; Piolot, Tristan; Dumoux, Maud; Vesque, Christine; Pichard, Evelyne; Dang, Tien; Duvauchelle, Boris; Poirier, Françoise; Delacour, Delphine

    2016-01-01

    Coordination of ciliary beating is essential to ensure mucus clearance in the airway tract. The orientation and synchronization of ciliary motion responds in part to the organization of the underlying cytoskeletal networks. Using electron tomography on mouse trachea, we show that basal bodies are collectively hooked at the cortex by a regular microtubule array composed of 4-5 microtubules. Removal of Galectin-3, one of basal body components, provokes misrecruitment of γ-tubulin, disorganization of this microtubule framework emanating from the basal foot cap, together with loss of basal body alignment and cilium orientation, defects in cilium organization and reduced fluid flow in the tracheal lumen. We conclude that Galectin-3 plays a crucial role in the maintenance of the microtubule organizing center of the cilium and the “pillar” microtubules, and that this network is instrumental for the coordinated orientation and stabilization of motile cilia. PMID:25215410

  11. Equilibrium shape of liquid lenses and correlations of beating cilia on the surface of frog embryos

    NASA Astrophysics Data System (ADS)

    Huisman, Fawn Mitsu

    This thesis reports the results of two unrelated projects: liquid lenses and cilia motion. A liquid lens is the equilibrium configuration of a non-wetting 3-fluid system, and the profile of the 3 interfaces can, in principle, be determined by solving the Young-Laplace differential equation for each interface if the surface tensions of all three interfaces are accurately known. Fluid interfacial tensions are not tabulated and are spectacularly sensitive to impurities, so in practice these quantities must be measured. We have developed a method of measuring these parameters by comparing the observed shape of liquid lenses to calculated equilibrium shapes. We have experimentally studied lenses of alkanes floating on water. These results were compared with various liquid lens systems. The profile of these lens systems was measured using ray tracing and Morie imaging, and found to be in excellent agreement with the calculated lens shapes for pure fluids. Upon the introduction of a surfactant, dodecyltrimethylammonium bromide(DTAB), we find that there is a minimum in the dihedral angle of the lens as a function of surfactant concentration, corresponding to the transition from partial-wetting to pseudo-partial wetting. The development and orientation of ciliated cells on Xenopus laevis larval skin was examined using high speed video microscopy. The intercellular orientation of wild-type, dominant negative mutants, and Vangl2MO morpholinos was studied by scoring the beating direction of ciliated cells and measuring the correlation with nearest neighbors. No significant difference between the mutant and wild type was found. Time lapse videos of developing cilia indicate that intracellular ordering is non-existent in early development, with ordering occurring by maturity. Further work needs to be done to determine what role fluid flow plays in that ordering.

  12. Mechanical Properties of a Primary Cilium Measured by Resonant Oscillation

    NASA Astrophysics Data System (ADS)

    Resnick, Andrew

    Primary cilia are ubiquitous mammalian cellular substructures implicated in an ever-increasing number of regulatory pathways. The well-established `ciliary hypothesis' states that physical bending of the cilium (for example, due to fluid flow) initiates signaling cascades, yet the mechanical properties of the cilium remain incompletely measured, resulting in confusion regarding the biological significance of flow-induced ciliary mechanotransduction. In this work we measure the mechanical properties of a primary cilium by using an optical trap to induce resonant oscillation of the structure. Our data indicate 1), the primary cilium is not a simple cantilevered beam, 2), the base of the cilium may be modeled as a nonlinear rotatory spring, the linear spring constant `k' of the cilium base calculated to be (4.6 +/- 0.62)*10-12 N/rad and nonlinear spring constant ` α' to be (-1 +/- 0.34) *10-10 N/rad2 , and 3) the ciliary base may be an essential regulator of mechanotransduction signalling. Our method is also particularly suited to measure mechanical properties of nodal cilia, stereocilia, and motile cilia, anatomically similar structures with very different physiological functions.

  13. Contiguous polarisation spectra of the Earth from 300 to 850 nm measured by GOME-2 onboard MetOp-A

    NASA Astrophysics Data System (ADS)

    Tilstra, L. G.; Lang, R.; Munro, R.; Aben, I.; Stammes, P.

    2014-07-01

    In this paper we present the first contiguous high-resolution spectra of the Earth's polarisation observed by a satellite instrument. The measurements of the Stokes fraction Q/I are performed by the spectrometer GOME-2 onboard the MetOp-A satellite. Polarisation measurements by GOME-2 are performed by onboard polarisation measurement devices (PMDs) and the high-resolution measurements discussed in this paper are taken in the special "PMD RAW" mode of operation. The spectral resolution of these PMD RAW polarisation measurements varies from 3 nm in the ultraviolet (UV) to 35 nm in the near-infrared wavelength range. We first compare measurements of the polarisation from cloud-free scenes with radiative transfer calculations for a number of cases. We find good agreement but also a spectral discrepancy at 800 nm, which we attribute to remaining imperfections in the calibration key data. Secondly, we study the polarisation of scenes with special scattering geometries that normally lead to near-zero Q/I. The GOME-2 polarisation spectra indeed show this behaviour and confirm the existence of the small discrepancy found earlier. Thirdly, we study the Earth polarisation for a variety of scenes. This provides a blueprint of Q/I over land and sea surfaces for various degrees of cloud cover. Fourthly, we compare the spectral dependence of measurements of Q/I in the UV with the generalised distribution function proposed by Schutgens and Stammes (2002) to describe the shape of the UV polarisation spectrum. The GOME-2 data confirm that these functions match the spectral behaviour captured by the GOME-2 PMD RAW mode.

  14. Contiguous polarisation spectra of the Earth from 300-850 nm measured by GOME-2 onboard MetOp-A

    NASA Astrophysics Data System (ADS)

    Tilstra, L. G.; Lang, R.; Munro, R.; Aben, I.; Stammes, P.

    2013-12-01

    In this paper we present the first contiguous high-resolution spectra of the Earth's polarisation observed by a satellite instrument. The measurements of the Stokes fraction Q/I are performed by the spectrometer GOME-2 onboard the MetOp-A satellite. Polarisation measurements by GOME-2 are performed by onboard polarisation measurement devices (PMDs) and the high-resolution measurements discussed in this paper are taken in the special "PMD RAW" mode of operation. The spectral resolution of these PMD RAW polarisation measurements varies from 3 nm in the ultraviolet (UV) to 35 nm in the near-infrared wavelength range. We first compare measurements of the polarisation from cloud-free scenes with radiative transfer calculations for a number of cases. We find good agreement but also a spectral discrepancy at 800 nm, which we attribute to remaining imperfections in the calibration key data. Secondly, we study the polarisation of scenes with special scattering geometries that normally lead to near-zero Q/I. The GOME-2 polarisation spectra indeed show this behaviour and confirm the existence of the small discrepancy found earlier. Thirdly, we study the Earth polarisation for a variety of scenes. This provides a blueprint of Q/I over land and sea surfaces for various degrees of cloud cover. Fourthly, we compare the spectral dependence of measurements of Q/I in the UV with the generalised distribution function that was proposed in the past (Schutgens and Stammes, 2002) to describe the shape of the UV polarisation spectrum. The GOME-2 data confirm that these functions match the spectral behaviour captured by the GOME-2 PMD RAW mode.

  15. Dual-Polarised Doppler X-band Radar Observations of Mixed Phased Clouds from the UK's Ice in Clouds Experiment-Dust (ICE-D)

    NASA Astrophysics Data System (ADS)

    Neely, Ryan; Blyth, Alan; Bennett, Lindsay; Dufton, David; Cui, Zhiqiang; McQuaid, Jim; Price, Hannah; Murray, Benjamin; Huang, Yahui

    2016-04-01

    Here we present dual-polarised X-band radar and in situ observations of convective, altocumulus and altostratus clouds relatively close to the Sahara desert in order to examine the impact of dust on the formation of ice and precipitation. These initial results come the UK's Ice in Clouds Experiment - Dust (UK ICE-D). UK ICE-D was an aircraft and ground-based project based in Cape Verde off the coast of Senegal, Africa during August 2015. The overall goal of this experiment was to determine how desert dust affects primary nucleation of ice particles in convective and layer clouds as well as the subsequent development of precipitation and glaciation of the clouds. This was accomplished by making focused observations when dust was present in high concentrations and when almost no dust was present. Here we focus on examining the differences in hydrometeor types derived from the dual-polarised X-band radar observations observed in the high and low dust loadings with specific emphasis on the role of supercooled rain drops in these two situations.

  16. Dual-Polarised Doppler X-band Radar Observations of Mixed Phased Clouds from the UK's Ice in Clouds Experiment-Dust (ICE-D)

    NASA Astrophysics Data System (ADS)

    Neely, R. R., III; Blyth, A. M.; Bennett, L.; Dufton, D.; Cui, Z.; Huang, Y.

    2015-12-01

    Here we present dual-polarised Doppler X-band radar observations of convective, altocumulus and altostratus clouds relatively close to the Sahara desert in order to examine the impact of dust on the formation of ice and precipitation. These initial results come the UK's Ice in Clouds Experiment - Dust (UK ICE-D). UK ICE-D was an aircraft and ground-based project based in Cape Verde off the coast of Senegal, Africa during August 2015. The overall goal of this experiment was to determine how desert dust affects primary nucleation of ice particles in convective and layer clouds as well as the subsequent development of precipitation and glaciation of the clouds. This was accomplished by making focused observations when dust was present in high concentrations and when almost no dust was present. Here we focus on examining the differences in hydrometeor types derived from the dual-polarised X-band radar observations observed in the high and low dust loadings with specific emphasis on the role of supercooled rain drops in these two situations.

  17. Immersed boundary-lattice Boltzmann method for simulation of muco-ciliary transport: effect of mucus depth at various amounts of cilia beat frequency

    NASA Astrophysics Data System (ADS)

    Shahmardan, M. M.; Sedaghat, M. H.; Norouzi, M.; Nazari, M.

    2015-12-01

    Numerical simulation based on immersed boundary-lattice Boltzmann method has been employed to study 2D muco-ciliary transport problem. The periciliary liquid (PCL) and mucus layers in this study are considered as the Newtonian and viscoelastic fluid respectively. An Oldroyd-B model is used as the constitutive equations of mucus layer. To simulate accurate effects of the cilia and PCL-mucus interface on the fluid, immersed boundary method is used. Numerical simulations have been performed to investigate the effects of mucus depth on the muco-ciliary clearance at various values of cilia beat frequencies. Our results show that, by increasing mucus depth, which results from air pollution and smoking, mean mucus velocity decreases. But it can be completely modified by increasing cilia beat frequency and the cilia beat frequency has great effect on the muco-ciliary clearance.

  18. Polycystins 1 and 2 mediate mechanosensation in the primary cilium of kidney cells

    NASA Technical Reports Server (NTRS)

    Nauli, Surya M.; Alenghat, Francis J.; Luo, Ying; Williams, Eric; Vassilev, Peter; Li, Xiaogang; Elia, Andrew E H.; Lu, Weining; Brown, Edward M.; Quinn, Stephen J.; Ingber, Donald E.; Zhou, Jing

    2003-01-01

    Several proteins implicated in the pathogenesis of polycystic kidney disease (PKD) localize to cilia. Furthermore, cilia are malformed in mice with PKD with mutations in TgN737Rpw (encoding polaris). It is not known, however, whether ciliary dysfunction occurs or is relevant to cyst formation in PKD. Here, we show that polycystin-1 (PC1) and polycystin-2 (PC2), proteins respectively encoded by Pkd1 and Pkd2, mouse orthologs of genes mutated in human autosomal dominant PKD, co-distribute in the primary cilia of kidney epithelium. Cells isolated from transgenic mice that lack functional PC1 formed cilia but did not increase Ca(2+) influx in response to physiological fluid flow. Blocking antibodies directed against PC2 similarly abolished the flow response in wild-type cells as did inhibitors of the ryanodine receptor, whereas inhibitors of G-proteins, phospholipase C and InsP(3) receptors had no effect. These data suggest that PC1 and PC2 contribute to fluid-flow sensation by the primary cilium in renal epithelium and that they both function in the same mechanotransduction pathway. Loss or dysfunction of PC1 or PC2 may therefore lead to PKD owing to the inability of cells to sense mechanical cues that normally regulate tissue morphogenesis.

  19. Inhibition of Autophagy Suppresses Sertraline-Mediated Primary Ciliogenesis in Retinal Pigment Epithelium Cells

    PubMed Central

    Kim, Eun Sung; Shin, Ji Hyun; Park, So Jung; Jo, Yoon Kyung; Kim, Jae-Sung; Kang, Il-Hwan; Nam, Jung-Bum; Chung, Doo-Young; Cho, Yoonchul; Lee, EunJoo H.; Chang, Jong Wook; Cho, Dong-Hyung

    2015-01-01

    Primary cilia are conserved cellular organelles that regulate diverse signaling pathways. Autophagy is a complex process of cellular degradation and recycling of cytoplasmic proteins and organelles, and plays an important role in cellular homeostasis. Despite its potential importance, the role of autophagy in ciliogenesis is largely unknown. In this study, we identified sertraline as a regulator of autophagy and ciliogenesis. Sertraline, a known antidepressant, induced the growth of cilia and blocked the disassembly of cilia in htRPE cells. Following treatment of sertraline, there was an increase in the number of cells with autophagic puncta and LC3 protein conversion. In addition, both a decrease of ATG5 expression and the treatment of an autophagy inhibitor resulted in the suppression of the sertraline-induced activation of autophagy in htRPE cells. Interestingly, we found that genetic and chemical inhibition of autophagy attenuated the growth of primary cilia in htRPE cells. Taken together, our results suggest that the inhibition of autophagy suppresses sertraline-induced ciliogenesis. PMID:25671433

  20. Qualitative Analysis of Collagen Fibers in Oral Submucous Fibrosis using Picrosirius Red Stain and Polarising Microscope

    PubMed Central

    Sekaran, Preethi; Narasimhan, Malathi

    2016-01-01

    Introduction Oral Submucous Fibrosis (OSMF) is an oral potentially malignant condition caused predominantly by areca nut chewing. Early recognition with accurate staging of the disease and appropriate treatment planning is of utmost importance to prevent the malignant transformation and to improve the quality of life of the patient. Picrosirius red stain is specific for collagen and enhances its birefringence under polarising light producing different colours in different stages of the disease. Aim To compare the clinical and functional staging with histopathologic staging methods used to assess the severity of OSMF and to perform a qualitative analysis of the collagen fibres in various histopathologic stages of OSMF using picrosirius red stain and polarising microscope. Materials and Methods It is a retrospective study done on archival samples. The study sample included a total of 30 cases which was divided into two groups. Group I comprised of 20 OSMF samples and Group II comprised of 10 normal tissue samples. Clinical, functional and histopathological staging was performed for all OSMF samples. Comparative analysis between clinical and functional stages with the histopathological staging was done using chi square test. Picrosirius red- stained sections of OSMF were analysed using polarising microscopy to evaluate the qualitative changes in the collagen fibers. Statistical Analysis Used Descriptive data which includes frequency and percentages were calculated for each group. Categorical data were analysed by chi-square test. A p-value of 0.05 or less was considered statistically significant. Results Comparative analysis between clinical and functional stages with the histopathological staging revealed a significant correlation (p < 0.05) between the functional and histopathological stage. Enhanced birefringence of the collagen fibers due to picrosirius red stain yielded characteristic prominent polarising colours in different stages of OSMF. Conclusion Comparison

  1. Physical Interpretation of the Sensitivity of Polarisation Coherence to Soil Surface Roughness

    NASA Astrophysics Data System (ADS)

    Mattia, F.; Le Toan, T.

    2003-04-01

    Surface roughness is an important geo-physical parameter required for numerous applications such as agronomy, geology, risk assessment, etc. In addition, the estimate of soil roughness may provide valuable a priori information to simplify the problem of soil moisture retrieval from SAR data. In the past, roughness discriminators based on the ratio between soil backscatter at different polarisations (i.e. sigmaHH/ sigmaVV ) and on the correlation coefficient between HH and VV channels (i.e. rhoHHVV) have been suggested. More recently, the potential of the correlation coefficient between co-polarised channels (i.e. polarisation coherence) in an arbitrary state of polarisation has been investigated. In particular, the correlation coefficient between co-polarised channels at circular polarisation (i.e. rhoRRLL ) has been found extremely sensitive to surface roughness and weakly sensitive to soil moisture content. However, notwithstanding these observations have been confirmed by several experimental studies a complete physical understanding of the phenomenon is still missing, at least in the remote sensing community. One of the main reasons for this lack of understanding is that in general, only lowest order approximations of theoretical surface scattering models are exploited in remote sensing applications. These approximations do not include the effect of multiple reflections. They cannot therefore predict accurately the whole covariance matrix often required to synthesise roughness discriminators, such as rhoRRLL. In this respect, despite the fact that higher order approximations of theoretical surface scattering models are mathematically very complex, they are necessary to give indications to understand the phenomenon and they can provide physical guidelines to develop semi-empirical approaches. In this context, the objective of this paper is to present a simple physical framework to interpret the sensitivity of different roughness discriminators to soil roughness

  2. Caenorhabditis elegans DYF-11, an orthologue of mammalian Traf3ip1/MIP-T3, is required for sensory cilia formation.

    PubMed

    Kunitomo, Hirofumi; Iino, Yuichi

    2008-01-01

    Cilia and flagella play critical roles in cell motility, development and sensory perception in animals. Formation and maintenance of cilia require a conserved protein transport system called intraflagellar transport (IFT). Here, we show that Caenorhabditis elegans dyf-11 encodes an evolutionarily conserved protein required for cilium biogenesis. dyf-11 is expressed in most of the ciliated neurons and is regulated by DAF-19, a crucial transcription factor for ciliary genes in C. elegans. dyf-11 mutants exhibit stunted cilia, fluorescent dye-filling defects (Dyf) of sensory neurons, and abnormal chemotaxis (Che). Cell- and stage-specific rescue experiments indicated that DYF-11 is required for formation and maintenance of sensory cilia in cell-autonomous manner. Fluorescent protein-tagged DYF-11 localizes to cilia and moves antero- and retrogradely via IFT. Analysis of DYF-11 movement in bbs mutants further suggested that DYF-11 is likely associated with IFT complex B. Domain analysis using DYF-11 deletion constructs revealed that the coiled-coil region is required for proper localization and ciliogenesis. We further show that Traf3ip1/MIP-T3, the mammalian orthologue of DYF-11, localizes to cilia in the MDCK renal epithelial cells. PMID:18173744

  3. Breakup and then makeup: a predictive model of how cilia self-regulate hardness for posture control

    NASA Astrophysics Data System (ADS)

    Bandyopadhyay, Promode R.; Hansen, Joshua C.

    2013-06-01

    Functioning as sensors and propulsors, cilia are evolutionarily conserved organelles having a highly organized internal structure. How a paramecium's cilium produces off-propulsion-plane curvature during its return stroke for symmetry breaking and drag reduction is not known. We explain these cilium deformations by developing a torsional pendulum model of beat frequency dependence on viscosity and an olivo-cerebellar model of self-regulation of posture control. The phase dependence of cilia torsion is determined, and a bio-physical model of hardness control with predictive features is offered. Crossbridge links between the central microtubule pair harden the cilium during the power stroke; this stroke's end is a critical phase during which ATP molecules soften the crossbridge-microtubule attachment at the cilium inflection point where torsion is at its maximum. A precipitous reduction in hardness ensues, signaling the start of ATP hydrolysis that re-hardens the cilium. The cilium attractor basin could be used as reference for perturbation sensing.

  4. Leading order determination of the gluon polarisation from DIS events with high-pT hadron pairs

    NASA Astrophysics Data System (ADS)

    Adolph, C.; Alekseev, M. G.; Alexakhin, V. Yu.; Alexandrov, Yu.; Alexeev, G. D.; Amoroso, A.; Antonov, A. A.; Austregesilo, A.; Badełek, B.; Balestra, F.; Barth, J.; Baum, G.; Bedfer, Y.; Bernhard, J.; Bertini, R.; Bettinelli, M.; Bicker, K.; Bieling, J.; Birsa, R.; Bisplinghoff, J.; Bordalo, P.; Bradamante, F.; Braun, C.; Bravar, A.; Bressan, A.; Burtin, E.; Chaberny, D.; Chiosso, M.; Chung, S. U.; Cicuttin, A.; Crespo, M. L.; Dalla Torre, S.; Das, S.; Dasgupta, S. S.; Denisov, O. Yu.; Dhara, L.; Donskov, S. V.; Doshita, N.; Duic, V.; Dünnweber, W.; Dziewiecki, M.; Efremov, A.; Elia, C.; Eversheim, P. D.; Eyrich, W.; Faessler, M.; Ferrero, A.; Filin, A.; Finger, M.; Finger, M.; Fischer, H.; Franco, C.; du Fresne von Hohenesche, N.; Friedrich, J. M.; Garfagnini, R.; Gautheron, F.; Gavrichtchouk, O. P.; Gazda, R.; Gerassimov, S.; Geyer, R.; Giorgi, M.; Gnesi, I.; Gobbo, B.; Goertz, S.; Grabmüller, S.; Grasso, A.; Grube, B.; Gushterski, R.; Guskov, A.; Guthörl, T.; Haas, F.; von Harrach, D.; Hedicke, S.; Heinsius, F. H.; Herrmann, F.; Heß, C.; Hinterberger, F.; Horikawa, N.; Höppner, Ch.; d'Hose, N.; Huber, S.; Ishimoto, S.; Ivanov, O.; Ivanshin, Yu.; Iwata, T.; Jahn, R.; Jasinski, P.; Joosten, R.; Kabuß, E.; Kang, D.; Ketzer, B.; Khaustov, G. V.; Khokhlov, Yu. A.; Kisselev, Yu.; Klein, F.; Klimaszewski, K.; Koblitz, S.; Koivuniemi, J. H.; Kolosov, V. N.; Kondo, K.; Königsmann, K.; Konorov, I.; Konstantinov, V. F.; Korzenev, A.; Kotzinian, A. M.; Kouznetsov, O.; Krämer, M.; Kroumchtein, Z. V.; Kunne, F.; Kurek, K.; Lauser, L.; Le Goff, J.-M.; Lednev, A. A.; Lehmann, A.; Levorato, S.; Lichtenstadt, J.; Maggiora, A.; Magnon, A.; Makke, N.; Mallot, G. K.; Mann, A.; Marchand, C.; Martin, A.; Marzec, J.; Matsuda, T.; Meyer, W.; Michigami, T.; Mikhailov, Yu. V.; Moinester, M. A.; Morreale, A.; Mutter, A.; Nagaytsev, A.; Nagel, T.; Nassalski, J. P.; Nerling, F.; Neubert, S.; Neyret, D.; Nikolaenko, V. I.; Nowak, W. D.; Nunes, A. S.; Olshevsky, A. G.; Ostrick, M.; Padee, A.; Panknin, R.; Panzieri, D.; Parsamyan, B.; Paul, S.; Perevalova, E.; Pesaro, G.; Peshekhonov, D. V.; Piragino, G.; Platchkov, S.; Pochodzalla, J.; Polak, J.; Polyakov, V. A.; Pontecorvo, G.; Pretz, J.; Procureur, S. L.; Quaresma, M.; Quintans, C.; Rajotte, J.-F.; Ramos, S.; Rapatsky, V.; Reicherz, G.; Richter, A.; Rocco, E.; Rondio, E.; Rossiyskaya, N. S.; Ryabchikov, D. I.; Samoylenko, V. D.; Sandacz, A.; Sapozhnikov, M. G.; Sarkar, S.; Savin, I. A.; Sbrizzai, G.; Schiavon, P.; Schill, C.; Schlüter, T.; Schmidt, K.; Schmitt, L.; Schönning, K.; Schopferer, S.; Schott, M.; Shevchenko, O. Yu.; Silva, L.; Sinha, L.; Sissakian, A. N.; Slunecka, M.; Smirnov, G. I.; Sosio, S.; Sozzi, F.; Srnka, A.; Stolarski, M.; Sulc, M.; Sulej, R.; Sznajder, P.; Takekawa, S.; Ter Wolbeek, J.; Tessaro, S.; Tessarotto, F.; Tkatchev, L. G.; Uhl, S.; Uman, I.; Vandenbroucke, M.; Virius, M.; Vlassov, N. V.; Wang, L.; Windmolders, R.; Wiślicki, W.; Wollny, H.; Zaremba, K.; Zavertyaev, M.; Zemlyanichkina, E.; Ziembicki, M.; Zhuravlev, N.; Zvyagin, A.

    2013-01-01

    We present a determination of the gluon polarisation Δg / g in the nucleon, based on the longitudinal double-spin asymmetry of DIS events with Q2 > 1 (GeV / c) 2 including a pair of large transverse-momentum hadrons in the final state. The data were obtained by the COMPASS experiment at CERN using a 160 GeV/c polarised muon beam scattering off a polarised 6LiD target. The gluon polarisation is evaluated by a Neural Network approach for three intervals of the gluon momentum fraction xg covering the range 0.04

  5. Coupled Hartree-Fock calculations of atomic polar tensors and the dipole polarisability of the benzene molecule

    NASA Astrophysics Data System (ADS)

    Lazzeretti, P.; Malagoli, M.; Zanasi, R.

    1990-03-01

    Coupled Hartree-Fock perturbation theory has been applied within the framework of high-quality basis sets to estimate the Hartree-Fock limit for the nuclear electric shielding tensors and the electric polarisability of the benzene molecule.

  6. Up to date on primary ciliary dyskinesia in children.

    PubMed

    Pifferi, Massimo; Di Cicco, Maria; Piras, Martina; Cangiotti, Angela M; Saggese, Giuseppe

    2013-10-01

    Primary ciliary dyskinesia (PCD) is a congenital, clinically and ultrastructurally heterogeneous disease due to abnormal structure and/or function of cilia, with impaired mucociliary transport leading to several respiratory disorders. PCD can be diagnosed by the combination of thorough clinical examination with functional and ultrastructural analysis of the cilia. This paper shows progresses in PCD diagnosis obtained by ciliogenesis in culture evaluation of ciliated respiratory cells and by genetic analysis of mutations in candidate genes. Moreover, since to date no specific treatments are available to correct the ciliary dysfunction, the paper shows the proper therapeutical approach by the use of respiratory physiotherapy and regular exercise to favour airways clearance, by antibiotics administration to control acute airway infections. Macrolides administration as antiinflammatory option is suggested. PMID:23973004

  7. Measurement of top quark polarisation in t-channel single top quark production

    SciTech Connect

    Khachatryan, Vardan

    2015-11-09

    Our first measurement of the top quark spin asymmetry, sensitive to the top quark polarisation, in t-channel single top quark production is presented. It is based on a sample of pp collisions at a centre-of-mass energy of 8 TeV corresponding to an integrated luminosity of 19.7 fb-1. A high-purity sample of t-channel single top quark events with an isolated muon is selected. Signal and background components are estimated using a fit to data. Furthermore, a differential cross section measurement, corrected for detector effects, of an angular observable sensitive to the top quark polarisation is performed. The differential distribution is used to extract a top quark spin asymmetry of 0.26 ± 0.03 (stat) ± 0.10 (syst), which is compatible with a p-value of 4.6% with the standard model prediction of 0.44.

  8. Coherent propagation of a short polarised radiation pulse in a one-dimensional resonance Bragg grating

    SciTech Connect

    Maimistov, Andrei I; Polikarpov, V V

    2006-09-30

    The propagation of an optical ultrashort pulse in a resonance Bragg grating is considered taking into account the polarisation of electromagnetic radiation. It is assumed that the grating is formed by thin films containing two-level atoms with the triply degenerate upper energy level. The system of equations is derived for the envelopes of electromagnetic pulses counterpropagating in such a grating. In the long-wavelength (continual) approximation, the system of equations generalising the known system for scalar waves is obtained. The solutions corresponding to elliptically (in particular, linearly and circularly) polarised stationary pulses are found. An arbitrary degree of ellipticity is possible only in a medium with a preliminary prepared stage of resonance atoms. (nonlinear optical phenomena)

  9. A new simple concept for ocean colour remote sensing using parallel polarisation radiance.

    PubMed

    He, Xianqiang; Pan, Delu; Bai, Yan; Wang, Difeng; Hao, Zengzhou

    2014-01-01

    Ocean colour remote sensing has supported research on subjects ranging from marine ecosystems to climate change for almost 35 years. However, as the framework for ocean colour remote sensing is based on the radiation intensity at the top-of-atmosphere (TOA), the polarisation of the radiation, which contains additional information on atmospheric and water optical properties, has largely been neglected. In this study, we propose a new simple concept to ocean colour remote sensing that uses parallel polarisation radiance (PPR) instead of the traditional radiation intensity. We use vector radiative transfer simulation and polarimetric satellite sensing data to demonstrate that using PPR has two significant advantages in that it effectively diminishes the sun glint contamination and enhances the ocean colour signal at the TOA. This concept may open new doors for ocean colour remote sensing. We suggest that the next generation of ocean colour sensors should measure PPR to enhance observational capability. PMID:24434904

  10. Polarised Multiangular Reflectance Measurements Using the Finnish Geodetic Institute Field Goniospectrometer

    PubMed Central

    Suomalainen, Juha; Hakala, Teemu; Peltoniemi, Jouni; Puttonen, Eetu

    2009-01-01

    The design, operation, and properties of the Finnish Geodetic Institute Field Goniospectrometer (FIGIFIGO) are presented. FIGIFIGO is a portable instrument for the measurement of surface Bidirectional Reflectance Factor (BRF) for samples with diameters of 10 – 50 cm. A set of polarising optics enable the measurement of linearly polarised BRF over the full solar spectrum (350 – 2,500 nm). FIGIFIGO is designed mainly for field operation using sunlight, but operation in a laboratory environment is also possible. The acquired BRF have an accuracy of 1 – 5% depending on wavelength, sample properties, and measurement conditions. The angles are registered at accuracies better than 2°. During 2004 – 2008, FIGIFIGO has been used in the measurement of over 150 samples, all around northern Europe. The samples concentrate mostly on boreal forest understorey, snow, urban surfaces, and reflectance calibration surfaces. PMID:22412342

  11. Measurement of top quark polarisation in t-channel single top quark production

    NASA Astrophysics Data System (ADS)

    Khachatryan, V.; Sirunyan, A. M.; Tumasyan, A.; Adam, W.; Asilar, E.; Bergauer, T.; Brandstetter, J.; Brondolin, E.; Dragicevic, M.; Erö, J.; Flechl, M.; Friedl, M.; Frühwirth, R.; Ghete, V. M.; Hartl, C.; Hörmann, N.; Hrubec, J.; Jeitler, M.; Knünz, V.; König, A.; Krammer, M.; Krätschmer, I.; Liko, D.; Matsushita, T.; Mikulec, I.; Rabady, D.; Rahbaran, B.; Rohringer, H.; Schieck, J.; Schöfbeck, R.; Strauss, J.; Treberer-Treberspurg, W.; Waltenberger, W.; Wulz, C.-E.; Mossolov, V.; Shumeiko, N.; Suarez Gonzalez, J.; Alderweireldt, S.; Cornelis, T.; de Wolf, E. A.; Janssen, X.; Knutsson, A.; Lauwers, J.; Luyckx, S.; van de Klundert, M.; van Haevermaet, H.; van Mechelen, P.; van Remortel, N.; van Spilbeeck, A.; Abu Zeid, S.; Blekman, F.; D'Hondt, J.; Daci, N.; de Bruyn, I.; Deroover, K.; Heracleous, N.; Keaveney, J.; Lowette, S.; Moreels, L.; Olbrechts, A.; Python, Q.; Strom, D.; Tavernier, S.; van Doninck, W.; van Mulders, P.; van Onsem, G. P.; van Parijs, I.; Barria, P.; Brun, H.; Caillol, C.; Clerbaux, B.; de Lentdecker, G.; Fasanella, G.; Favart, L.; Grebenyuk, A.; Karapostoli, G.; Lenzi, T.; Léonard, A.; Maerschalk, T.; Marinov, A.; Perniè, L.; Randle-Conde, A.; Seva, T.; Vander Velde, C.; Vanlaer, P.; Yonamine, R.; Zenoni, F.; Zhang, F.; Beernaert, K.; Benucci, L.; Cimmino, A.; Crucy, S.; Dobur, D.; Fagot, A.; Garcia, G.; Gul, M.; McCartin, J.; Ocampo Rios, A. A.; Poyraz, D.; Ryckbosch, D.; Salva, S.; Sigamani, M.; Tytgat, M.; van Driessche, W.; Yazgan, E.; Zaganidis, N.; Basegmez, S.; Beluffi, C.; Bondu, O.; Brochet, S.; Bruno, G.; Caudron, A.; Ceard, L.; da Silveira, G. G.; Delaere, C.; Favart, D.; Forthomme, L.; Giammanco, A.; Hollar, J.; Jafari, A.; Jez, P.; Komm, M.; Lemaitre, V.; Mertens, A.; Musich, M.; Nuttens, C.; Perrini, L.; Pin, A.; Piotrzkowski, K.; Popov, A.; Quertenmont, L.; Selvaggi, M.; Vidal Marono, M.; Beliy, N.; Hammad, G. H.; Aldá Júnior, W. L.; Alves, F. L.; Alves, G. A.; Brito, L.; Correa Martins Junior, M.; Hamer, M.; Hensel, C.; Mora Herrera, C.; Moraes, A.; Pol, M. E.; Rebello Teles, P.; Belchior Batista Das Chagas, E.; Carvalho, W.; Chinellato, J.; Custódio, A.; da Costa, E. M.; de Jesus Damiao, D.; de Oliveira Martins, C.; Fonseca de Souza, S.; Huertas Guativa, L. M.; Malbouisson, H.; Matos Figueiredo, D.; Mundim, L.; Nogima, H.; Prado da Silva, W. L.; Santoro, A.; Sznajder, A.; Tonelli Manganote, E. J.; Vilela Pereira, A.; Ahuja, S.; Bernardes, C. A.; de Souza Santos, A.; Dogra, S.; Fernandez Perez Tomei, T. R.; Gregores, E. M.; Mercadante, P. G.; Moon, C. S.; Novaes, S. F.; Padula, Sandra S.; Romero Abad, D.; Ruiz Vargas, J. C.; Aleksandrov, A.; Hadjiiska, R.; Iaydjiev, P.; Rodozov, M.; Stoykova, S.; Sultanov, G.; Vutova, M.; Dimitrov, A.; Glushkov, I.; Litov, L.; Pavlov, B.; Petkov, P.; Ahmad, M.; Bian, J. G.; Chen, G. M.; Chen, H. S.; Chen, M.; Cheng, T.; Du, R.; Jiang, C. H.; Plestina, R.; Romeo, F.; Shaheen, S. M.; Spiezia, A.; Tao, J.; Wang, C.; Wang, Z.; Zhang, H.; Asawatangtrakuldee, C.; Ban, Y.; Li, Q.; Liu, S.; Mao, Y.; Qian, S. J.; Wang, D.; Xu, Z.; Avila, C.; Cabrera, A.; Chaparro Sierra, L. F.; Florez, C.; Gomez, J. P.; Gomez Moreno, B.; Sanabria, J. C.; Godinovic, N.; Lelas, D.; Puljak, I.; Ribeiro Cipriano, P. M.; Antunovic, Z.; Kovac, M.; Brigljevic, V.; Kadija, K.; Luetic, J.; Micanovic, S.; Sudic, L.; Attikis, A.; Mavromanolakis, G.; Mousa, J.; Nicolaou, C.; Ptochos, F.; Razis, P. A.; Rykaczewski, H.; Bodlak, M.; Finger, M.; Finger, M.; Abdelalim, A. A.; Awad, A.; El Sawy, M.; Mahrous, A.; Radi, A.; Calpas, B.; Kadastik, M.; Murumaa, M.; Raidal, M.; Tiko, A.; Veelken, C.; Eerola, P.; Pekkanen, J.; Voutilainen, M.; Härkönen, J.; Karimäki, V.; Kinnunen, R.; Lampén, T.; Lassila-Perini, K.; Lehti, S.; Lindén, T.; Luukka, P.; Mäenpää, T.; Peltola, T.; Tuominen, E.; Tuominiemi, J.; Tuovinen, E.; Wendland, L.; Talvitie, J.; Tuuva, T.; Besancon, M.; Couderc, F.; Dejardin, M.; Denegri, D.; Fabbro, B.; Faure, J. L.; Favaro, C.; Ferri, F.; Ganjour, S.; Givernaud, A.; Gras, P.; Hamel de Monchenault, G.; Jarry, P.; Locci, E.; Machet, M.; Malcles, J.; Rander, J.; Rosowsky, A.; Titov, M.; Zghiche, A.; Antropov, I.; Baffioni, S.; Beaudette, F.; Busson, P.; Cadamuro, L.; Chapon, E.; Charlot, C.; Dahms, T.; Davignon, O.; Filipovic, N.; Florent, A.; Granier de Cassagnac, R.; Jo, M.; Lisniak, S.; Mastrolorenzo, L.; Miné, P.; Naranjo, I. N.; Nguyen, M.; Ochando, C.; Ortona, G.; Paganini, P.; Pigard, P.; Regnard, S.; Salerno, R.; Sauvan, J. B.; Sirois, Y.; Strebler, T.; Yilmaz, Y.; Zabi, A.; Agram, J.-L.; Andrea, J.; Aubin, A.; Bloch, D.; Brom, J.-M.; Buttignol, M.; Chabert, E. C.; Chanon, N.; Collard, C.; Conte, E.; Coubez, X.; Fontaine, J.-C.; Gelé, D.; Goerlach, U.; Goetzmann, C.; Le Bihan, A.-C.; Merlin, J. A.; Skovpen, K.

    2016-04-01

    A first measurement of the top quark spin asymmetry, sensitive to the top quark polarisation, in t-channel single top quark production is presented. It is based on a sample of pp collisions at a centre-of-mass energy of 8 TeV corresponding to an integrated luminosity of 19.7 fb-1. A high-purity sample of t-channel single top quark events with an isolated muon is selected. Signal and background components are estimated using a fit to data. A differential cross section measurement, corrected for detector effects, of an angular observable sensitive to the top quark polarisation is performed. The differential distribution is used to extract a top quark spin asymmetry of 0.26 ± 0.03(stat) ± 0.10(syst), which is compatible with a p-value of 4.6% with the standard model prediction of 0.44. [Figure not available: see fulltext.

  12. Polarised interleukin 8 secretion by HT 29/19A cells.

    PubMed Central

    Lammers, K M; Jansen, J; Bijlsma, P B; Ceska, M; Tytgat, G N; Laboisse, C L; van Deventer, S J

    1994-01-01

    Interleukin 8 is a neutrophil chemotactic and stimulating cytokine induced by various inflammatory stimuli, including tumour necrosis factor, interleukin 1, and endotoxin. The ability of HT 29/19A enterocytes to synthesise interleukin 8 was studied. The results show that interleukin 1 is an important stimulus for interleukin 8 synthesis and secretion by HT 29/19A cells, being more potent than tumour necrosis factor. The tumour necrosis factor and interleukin 1 induced interleukin 8 secretion by HT 29/19A cells was seen to be polarised according to the direction of stimulation. These results support the concept that mucosal cells (enterocytes) may play an important part in initiating mucosal inflammation. Furthermore, it is proposed that HT 29/19A cells constitute a tool to study stimulus directed polarised cytokine secretion. Images Figure 3 PMID:8150343

  13. Measurement of top quark polarisation in t-channel single top quark production

    DOE PAGESBeta

    Khachatryan, Vardan

    2016-04-13

    Our first measurement of the top quark spin asymmetry, sensitive to the top quark polarisation, in t-channel single top quark production is presented. It is based on a sample of pp collisions at a centre-of-mass energy of 8 TeV corresponding to an integrated luminosity of 19.7 fb-1. A high-purity sample of t-channel single top quark events with an isolated muon is selected. Signal and background components are estimated using a fit to data. Furthermore, a differential cross section measurement, corrected for detector effects, of an angular observable sensitive to the top quark polarisation is performed. The differential distribution is usedmore » to extract a top quark spin asymmetry of 0.26 ± 0.03 (stat) ± 0.10 (syst), which is compatible with a p-value of 4.6% with the standard model prediction of 0.44.« less

  14. Gluon Polarisation in the Nucleon from High Transverse Momentum Hadron Pairs at COMPASS

    SciTech Connect

    Kurek, Krzysztof

    2009-08-04

    The main goal of the COMPASS spin physics programme is the measurement of the helicity contribution of the gluons, {delta}G/G, to the nucleon spin. It is determined from the longitudinal spin asymmetry in the scattering of 160 GeV polarised muons off a polarised LiD target. The new analysis and new COMPASS {delta}G/G result obtained from high transverse momentum hadron pairs for Q{sup 2}>1(GeV/c){sup 2} is presented. The weighted method based on a Neural Network approach is discussed. The result of {delta}G/G 0.08{+-}0.10(stat){+-}0.05(syst)disfavors a large positive contribution to the nucleon spin for a gluon momentum fraction around 0.08 and agrees with the previously obtained result from the analysis for the small Q{sup 2} kinematical region (Q{sup 2}<1(GeV/c){sup 2})

  15. Determination of Polarised Parton Distributions in the Nucleon --- Next to Leading Order QCD Analysis

    NASA Astrophysics Data System (ADS)

    Tatur, Stanislaw; Bartelski, Jan; Kurzela, Miroslaw

    2000-03-01

    We have made next to leading order QCD fit to the deep inelastic spin asymmetries on nucleons and we have determined polarised quark and gluon densities. The functional form for such distributions was inspired by the Martin, Roberts and Stirling fit for unpolarised case. In addition to usually used data points (averaged over x and Q2) we have also considered the sample containing points with similar x and different Q2. It seems that splitting of quark densities into valence and sea contribution is strongly model dependent and only their sum (i.e. , Δ u and Δ d) can be precisely determined from the data. Integrated polarised gluon contribution, contrary to some expectations, is relatively small and the sign of it depends on the fact which sample of data points is used.

  16. A new simple concept for ocean colour remote sensing using parallel polarisation radiance

    PubMed Central

    He, Xianqiang; Pan, Delu; Bai, Yan; Wang, Difeng; Hao, Zengzhou

    2014-01-01

    Ocean colour remote sensing has supported research on subjects ranging from marine ecosystems to climate change for almost 35 years. However, as the framework for ocean colour remote sensing is based on the radiation intensity at the top-of-atmosphere (TOA), the polarisation of the radiation, which contains additional information on atmospheric and water optical properties, has largely been neglected. In this study, we propose a new simple concept to ocean colour remote sensing that uses parallel polarisation radiance (PPR) instead of the traditional radiation intensity. We use vector radiative transfer simulation and polarimetric satellite sensing data to demonstrate that using PPR has two significant advantages in that it effectively diminishes the sun glint contamination and enhances the ocean colour signal at the TOA. This concept may open new doors for ocean colour remote sensing. We suggest that the next generation of ocean colour sensors should measure PPR to enhance observational capability. PMID:24434904

  17. Continuous-wave parametric oscillation in polarisation-maintaining optical fibre

    SciTech Connect

    Zlobina, E A; Kablukov, S I; Babin, Sergei A

    2011-09-30

    Continuous-wave parametric oscillation in a polarisation-maintaining optical fibre has been achieved for the first time using polarisation phase matching. Up-conversion with a frequency shift of {approx}8.6 THz has been demonstrated experimentally. The single-pass optical power generated at 1017 nm in a 85-m-long Nufern PM980-XP fibre exceeded 40 mW. The conversion efficiency was 3.3%, which is an order of magnitude higher than that reported previously for cw parametric up-conversion in the 1-{mu}m spectral region. We compare theoretical predictions with experimental data and analyse factors that may influence output power saturation with increasing signal and pump powers.

  18. A new simple concept for ocean colour remote sensing using parallel polarisation radiance

    NASA Astrophysics Data System (ADS)

    He, Xianqiang; Pan, Delu; Bai, Yan; Wang, Difeng; Hao, Zengzhou

    2014-01-01

    Ocean colour remote sensing has supported research on subjects ranging from marine ecosystems to climate change for almost 35 years. However, as the framework for ocean colour remote sensing is based on the radiation intensity at the top-of-atmosphere (TOA), the polarisation of the radiation, which contains additional information on atmospheric and water optical properties, has largely been neglected. In this study, we propose a new simple concept to ocean colour remote sensing that uses parallel polarisation radiance (PPR) instead of the traditional radiation intensity. We use vector radiative transfer simulation and polarimetric satellite sensing data to demonstrate that using PPR has two significant advantages in that it effectively diminishes the sun glint contamination and enhances the ocean colour signal at the TOA. This concept may open new doors for ocean colour remote sensing. We suggest that the next generation of ocean colour sensors should measure PPR to enhance observational capability.

  19. Biochemical studies of olfaction: binding specificity of odorants to a cilia preparation from rainbow trout olfactory rosettes.

    PubMed

    Rhein, L D; Cagan, R H

    1983-08-01

    Cilia isolated from the olfactory epithelium (olfactory rosettes) of rainbow trout (Salmo gairdneri) bind amino acids, which are odor stimuli to this species. We demonstrate that L-threonine, L-serine, and L-alanine bind to a common site, TSA, in the cilia preparation. All possible mixtures of two of the amino acids as competitors, with the third as the 3H-labeled ligand, were studied. The effect of two combined (unlabeled) competitors was always substantially less than additive compared with their actions singly. Along with additional inhibition studies using mixtures of inhibitors, the data show that the three odorants must interact with at least one common binding site, TSA. Binding of L-[3H]lysine to site L was unaffected by addition of L-threonine, L-serine, or L-alanine, establishing its independence from site TSA. L-Arginine inhibited binding of L-[3H]lysine, showing that both of these basic amino acids interact with site L. The data establish the presence, in trout olfactory cilia, of at least two separate and noninteracting populations of odorant binding sites, TSA and L. PMID:6409997

  20. VizieR Online Data Catalog: Polarisation of a sample of late M dwarfs (Morin+, 2010)

    NASA Astrophysics Data System (ADS)

    Morin, J.; Donati, J.-F.; Petit, P.; Delfosse, X.; Forveille, T.; Jardine, M. M.

    2010-06-01

    We have collected 174 pairs of Stokes I (unpolarised) and V (circularly polarised) spectra with ESPaDOnS at CFHT (2003ASPC..307...41D) between June 2006 and July 2009. All spectra were reduced using the Libre-Esprit pipeline, and the mean I and V line profiles were extracted using the Least-Squares Deconvolution (LSD) technique (Donati et al., 1997MNRAS.291..658D). (3 data files).

  1. Continuum optical circular polarisation in the young O star Theta 1 Orionis C?

    NASA Astrophysics Data System (ADS)

    Moffat, A. F. J.; Eversberg, T.

    2000-06-01

    Recently, Donati & Wade (1999) have claimed rather spectacular, large, variable circular polarisation in the optical continuum of Theta 1 Orionis C, obtained with the échelle spectropolarimeter MuSiCoS. However, based on experience with the William-Wehlau spectropolarimeter, a similar unit using two fiber feeds, we suggest that this is the spurious result of instrumental effects. We propose a remedy to eliminate the effect.

  2. Dust in the diffuse interstellar medium. Extinction, emission, linear and circular polarisation

    NASA Astrophysics Data System (ADS)

    Siebenmorgen, R.; Voshchinnikov, N. V.; Bagnulo, S.

    2014-01-01

    We present a model for the diffuse interstellar dust that explains the observed wavelength-dependence of extinction, emission, and the linear and circular polarisation of light. The model is set up with a small number of parameters. It consists of a mixture of amorphous carbon and silicate grains with sizes from the molecular domain of 0.5 up to about 500 nm. Dust grains with radii larger than 6 nm are spheroids. Spheroidal dust particles have a factor 1.5-3 greater absorption cross section in the far-infrared than spherical grains of the same volume do. Mass estimates derived from submillimetre observations that ignore this effect are overestimated by the same amount. In the presence of a magnetic field, spheroids may be partly aligned and polarise light. We find that polarisation spectra help to determine the upper particle radius of the otherwise rather unconstrained dust size distribution. Stochastically heated small grains of graphite, silicates, and polycyclic aromatic hydrocarbons (PAHs) are included. We tabulate parameters for PAH emission bands in various environments. They show a trend with the hardness of the radiation field that can be explained by the ionisation state or hydrogenation coverage of the molecules. For each dust component its relative weight is specified so that absolute element abundances are not direct input parameters. The model is compared to the average properties of the Milky Way, which seem to represent dust in the solar neighbourhood. It is then applied to specific sight lines towards four particular stars, with one of them located in the reflection nebula NGC 2023. For these sight lines, we present ultra-high signal-to-noise linear and circular spectro-polarimetric observations obtained with FORS at the VLT. Using prolate rather than oblate grains gives a better fit to observed spectra; the axial ratio of the spheroids is typically two and aligned silicates are the dominant contributors to the polarisation. Based on ESO: 386.C

  3. Static multipole polarisabilities and second-order Stark shift in francium.

    PubMed

    Khan, F; Khandelwal, G S; Wilson, J W

    1988-01-01

    The multipole polarisability of the ground state of francium is calculated by utilising both the variational technique of Davison and the quantum defect theory underlying the Bates-Damgaard method. This approach is also shown to yield reasonable results for other alkali atoms. Second-order Stark shift for the ground state of francium is presented as a function of field strength for possible future experimental comparison. PMID:11539071

  4. High-Accuracy Analysis of Compton Scattering in Chiral EFT: Proton and Neutron Polarisabilities

    NASA Astrophysics Data System (ADS)

    Griesshammer, Harald W.; Phillips, Daniel R.; McGovern, Judith A.

    2013-10-01

    Compton scattering from protons and neutrons provides important insight into the structure of the nucleon. A new extraction of the static electric and magnetic dipole polarisabilities αE 1 and βM 1 of the proton and neutron from all published elastic data below 300 MeV in Chiral Effective Field Theory shows that within the statistics-dominated errors, the proton and neutron polarisabilities are identical, i.e. no iso-spin breaking effects of the pion cloud are seen. Particular attention is paid to the precision and accuracy of each data set, and to an estimate of residual theoretical uncertainties. ChiEFT is ideal for that purpose since it provides a model-independent estimate of higher-order corrections and encodes the correct low-energy dynamics of QCD, including, for few-nucleon systems used to extract neutron polarisabilities, consistent nuclear currents, rescattering effects and wave functions. It therefore automatically respects the low-energy theorems for photon-nucleus scattering. The Δ (1232) as active degree of freedom is essential to realise the full power of the world's Compton data.Its parameters are constrained in the resonance region. A brief outlook is provided on what kind of future experiments can improve the database. Supported in part by UK STFC, DOE, NSF, and the Sino-German CRC 110.

  5. Reprint of : Majorana fermion fingerprints in spin-polarised scanning tunnelling microscopy

    NASA Astrophysics Data System (ADS)

    Kotetes, Panagiotis; Mendler, Daniel; Heimes, Andreas; Schön, Gerd

    2016-08-01

    We calculate the spatially resolved tunnelling conductance of topological superconductors (TSCs) based on ferromagnetic chains, measured by means of spin-polarised scanning tunnelling microscopy (SPSTM). Our analysis reveals novel signatures of MFs arising from the interplay of their strongly anisotropic spin-polarisation and the magnetisation content of the tip. We focus on the deep Yu-Shiba-Rusinov (YSR) limit where only YSR bound states localised in the vicinity of the adatoms govern the low-energy as also the topological properties of the system. Under these conditions, we investigate the occurrence of zero/finite bias peaks (ZBPs/FBPs) for a single or two coupled TSC chains forming a Josephson junction. Each TSC can host up to two Majorana fermions (MFs) per edge if chiral symmetry is preserved. Here we retrieve the conductance for all the accessible configurations of the MF number of each chain. Our results illustrate innovative spin-polarisation-sensitive experimental routes for arresting the MFs by either restoring or splitting the ZBP in a predictable fashion via: (i) weakly breaking chiral symmetry, e.g. by the SPSTM tip itself or by an external Zeeman field and (ii) tuning the superconducting phase difference of the TSCs, which is encoded in the 4π-Josephson coupling of neighbouring MFs.

  6. Counterintuitive electron localisation from density-functional theory with polarisable solvent models

    SciTech Connect

    Dale, Stephen G.; Johnson, Erin R.

    2015-11-14

    Exploration of the solvated electron phenomena using density-functional theory (DFT) generally results in prediction of a localised electron within an induced solvent cavity. However, it is well known that DFT favours highly delocalised charges, rendering the localisation of a solvated electron unexpected. We explore the origins of this counterintuitive behaviour using a model Kevan-structure system. When a polarisable-continuum solvent model is included, it forces electron localisation by introducing a strong energetic bias that favours integer charges. This results in the formation of a large energetic barrier for charge-hopping and can cause the self-consistent field to become trapped in local minima thus converging to stable solutions that are higher in energy than the ground electronic state. Finally, since the bias towards integer charges is caused by the polarisable continuum, these findings will also apply to other classical polarisation corrections, as in combined quantum mechanics and molecular mechanics (QM/MM) methods. The implications for systems beyond the solvated electron, including cationic DNA bases, are discussed.

  7. A New High-Accuracy Analysis of Compton Scattering in Chiral EFT: Neutron Polarisabilities

    NASA Astrophysics Data System (ADS)

    Griesshammer, Harald W.; McGovern, Judith A.; Phillips, Daniel R.

    2015-04-01

    Low-energy Compton scattering tests the symmetries and interaction strengths of a target's internal degrees of freedom in the electric and magnetic fields of a real, external photon. In the single-nucleon sector, information is often compressed into the static scalar dipole polarisabilities which are experimentally not directly accessible but encode information on the pion cloud and the Δ(1232) excitation. The interaction of the photon with the charged pion-exchange also provides a conceptually clean probe of few-nucleon binding. After demonstrating the statistical consistency of the world's γd dataset including the new data from the MAX-IV collaboration described in the preceding talk, we present a new extraction of the neutron polarisabilities in Chiral Effective Field Theory: αn = [ 11 . 55 +/- 1 . 25(stat) +/- 0 . 2(BSR) +/- 0 . 8(th) ] and βn = [ 3 . 65 -/+ 1 . 25(stat) +/- 0 . 2(BSR) -/+ 0 . 8(th) ] , in 10-4 fm3, with χ2 = 45 . 2 for 44 degrees of freedom. The new data reduced the statistical uncertainties by 30%. We discuss data accuracy and consistency, the role of the Δ(1232) , and an estimate of residual theoretical uncertainties. Within statistical and systematic errors, proton and neutron polarisabilities remain identical. Supported in part by UK STFC and US DOE.

  8. An improved simple polarisable water model for use in biomolecular simulation

    SciTech Connect

    Bachmann, Stephan J.; Gunsteren, Wilfred F. van

    2014-12-14

    The accuracy of biomolecular simulations depends to some degree on the accuracy of the water model used to solvate the biomolecules. Because many biomolecules such as proteins are electrostatically rather inhomogeneous, containing apolar, polar, and charged moieties or side chains, a water model should be able to represent the polarisation response to a local electrostatic field, while being compatible with the force field used to model the biomolecules or protein. The two polarisable water models, COS/G2 and COS/D, that are compatible with the GROMOS biomolecular force fields leave room for improvement. The COS/G2 model has a slightly too large dielectric permittivity and the COS/D model displays a much too slow dynamics. The proposed COS/D2 model has four interaction sites: only one Lennard-Jones interaction site, the oxygen atom, and three permanent charge sites, the two hydrogens, and one massless off-atom site that also serves as charge-on-spring (COS) polarisable site with a damped or sub-linear dependence of the induced dipole on the electric field strength for large values of the latter. These properties make it a cheap and yet realistic water model for biomolecular solvation.

  9. The Ringo2 Optical Polarisation Catalogue of 13 High-Energy Blazars

    NASA Astrophysics Data System (ADS)

    Barres de Almeida, Ulisses; Jermak, Helen; Mundell, Carole; Lindfors, Elina; Nilsson, Kari; Steele, Iain

    2015-08-01

    We present the findings of the Ringo2 3-year survey of 13 blazars (3 FSRQs and 10 BL Lacs) with regular coverage and reasonably fast cadence of one to three observations a week. Ringo2 was installed on the Liverpool Robotic Telescope (LT) on the Canary Island of La Palma between 2009 and 2012 and monitored thirteen high-energy-emitting blazars in the northern sky. The objects selected as well as the observational strategy were tuned to maximise the synergies with high-energy X- to gamma-ray observations. Therefore this sample stands out as a well-sampled, long-term view of high-energy AGN jets in polarised optical light. Over half of the sources exhibited an increase in optical flux during this period and almost a quarter were observed in outburst. We compare the optical data to gamma (Fermi/LAT) and X-ray data during these periods of outburst. In this talk we present the data obtained for all sources over the lifetime of Ringo2 with additional optical data from the KVA telescope and the SkyCamZ wide-field camera (on the LT), we explore the relationship between the change in polarisation angle as a function of time (dEVPA/dMJD), flux and polarisation degree along with cross correlation comparisons of optical and high-energy flux.

  10. Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory Neurons

    PubMed Central

    Boccaccio, Anna; Sagheddu, Claudia; Menini, Anna

    2011-01-01

    Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds1. Caged compounds are molecules made physiologically inactive by a chemical cage that can be broken by a flash of ultraviolet light. Here, we show how to obtain patch-clamp recordings combined with photolysis of caged compounds for the study of olfactory transduction in dissociated mouse olfactory sensory neurons. The process of olfactory transduction (Figure 1) takes place in the cilia of olfactory sensory neurons, where odorant binding to receptors leads to the increase of cAMP that opens cyclic nucleotide-gated (CNG) channels2. Ca entry through CNG channels activates Ca-activated Cl channels. We show how to dissociate neurons from the mouse olfactory epithelium3 and how to activate CNG channels or Ca-activated Cl channels by photolysis of caged cAMP4 or caged Ca5. We use a flash lamp6,7 to apply ultraviolet flashes to the ciliary region to uncage cAMP or Ca while patch-clamp recordings are taken to measure the current in the whole-cell voltage-clamp configuration8-11. PMID:22064384

  11. Emergence of polar order and cooperativity in hydrodynamically coupled model cilia

    PubMed Central

    Bruot, Nicolas; Cicuta, Pietro

    2013-01-01

    As a model of ciliary beat, we use two-state oscillators that have a defined direction of oscillation and have strong synchronization properties. By allowing the direction of oscillation to vary according to the interaction with the fluid, with a timescale longer than the timescale of synchronization, we show in simulations that several oscillators can align in a direction set by the geometrical configuration of the system. In this system, the alignment depends on the state of synchronization of the system, and is therefore linked to the beat pattern of the model cilia. By testing various configurations from two to 64 oscillators, we deduce empirically that, when the synchronization state of neighbouring oscillators is in phase, the angles of the oscillators align in a configuration of high hydrodynamic coupling. In arrays of oscillators that break the planar symmetry, a global direction of alignment emerges reflecting this polarity. In symmetric configurations, where several directions are geometrically equivalent, the array still displays strong internal cooperative behaviour. It also appears that the shape of the array is more important than the lattice type and orientation in determining the preferred direction. PMID:23883957

  12. Remodeling Cildb, a popular database for cilia and links for ciliopathies

    PubMed Central

    2014-01-01

    Background New generation technologies in cell and molecular biology generate large amounts of data hard to exploit for individual proteins. This is particularly true for ciliary and centrosomal research. Cildb is a multi–species knowledgebase gathering high throughput studies, which allows advanced searches to identify proteins involved in centrosome, basal body or cilia biogenesis, composition and function. Combined to localization of genetic diseases on human chromosomes given by OMIM links, candidate ciliopathy proteins can be compiled through Cildb searches. Methods Othology between recent versions of the whole proteomes was computed using Inparanoid and ciliary high throughput studies were remapped on these recent versions. Results Due to constant evolution of the ciliary and centrosomal field, Cildb has been recently upgraded twice, with new species whole proteomes and new ciliary studies, and the latter version displays a novel BioMart interface, much more intuitive than the previous ones. Conclusions This already popular database is designed now for easier use and is up to date in regard to high throughput ciliary studies. PMID:25422781

  13. Ionic control of the reversal response of cilia in Paramecium caudatum. A calcium hypothesis.

    PubMed

    Naitoh, Y

    1968-01-01

    The duration of ciliary reversal of Paramecium caudatum in response to changes in external ionic factors was determined with various ionic compositions of both equilibration and stimulation media. The reversal response was found to occur when calcium ions bound by an inferred cellular cation exchange system were liberated in exchange for externally applied cations other than calcium. Factors which affect the duration of the response were (a) initial amount of calcium bound by the cation exchange system, (b) final amount of calcium bound by the system after equilibration with the stimulation medium, and (c) concentration of calcium ions in the stimulation medium. An empirical equation is presented which relates the duration of the response to these three factors. On the basis of these and previously published data, the following hypothesis is proposed for the mechanism underlying ciliary reversal in response to cationic stimulation: Ca(++) liberated from the cellular cation exchange system activates a contractile system which is energized by ATP. Contraction of this component results in the reversal of effective beat direction of cilia by a mechanism not yet understood. The duration of reversal in live paramecia is related to the time course of bound calcium release. PMID:4966766

  14. Photolysis of caged calcium in cilia induces ciliary reversal in Paramecium caudatum.

    PubMed

    Iwadate, Yoshiaki

    2003-04-01

    Intracellular Ca(2+) concentration controls both the pattern and frequency of ciliary and flagellar beating in eukaryotes. In Paramecium, it is widely accepted that the reversal of the direction of ciliary beating (ciliary reversal) is induced by an increase in intra-ciliary Ca(2+) levels. Despite this, the Ca(2+)-sensitive region of the cilium that initiates ciliary reversal has not been clearly identified. We injected caged calcium into living P. caudatum cells and applied ultraviolet (UV) light to portions of the injected cells to raise artificially the intracellular Ca(2+) level ([Ca(2+)](i)). UV application to the upper ciliary region above the basal body induced ciliary reversal in injected cells. Furthermore, UV application to the tips of cilia induced weak ciliary reversal. Larger areas of photolysis in the cilium gave rise to greater angles of ciliary reversal. These results strongly suggest that the Ca(2+)-sensitive region for ciliary reversal is distributed all over the cilium, above the basal body. PMID:12604576

  15. [Correction of the position of the cilia in facial paralysis: Technical note].

    PubMed

    Caillot, A; Labbé, D

    2015-06-01

    Facial paralysis is a incapacitating pathology that we treat with lengthening temporalis myoplasty for reanimation of the smile. To treat lagophthalmia, we use the extension of the levator of the upper eyelid according Tessier and the asymmetric external blepharorraphy. These techniques can optionally be combined with other techniques, as needed. However, many patients are embarrassed by the appearance of the lashes of the upper eyelid homolateral side facial paralysis. The cilia are lowered and horizontalised, creating a functional disorder by partial "amputation" of the visual field and aesthetic inconvenience. We describe a surgical technique to correct the malposition of the lashes. This technique can be carried out independently or in the lengthening of the temporal myoplasty or another surgical procedure on the eye. In case of extension of the levator of the upper eyelid, the technique we propose requires no additional incision. This is a simple technique and increases very little surgical time. It is fast, little or no morbid, reproducible and provides a significant improvement in the aesthetic and functional patient. This simple technique allows to provide both aesthetic and functional refinement for patients with facial paralysis sequelae. PMID:25708730

  16. Self-organization in cytoskeletal mixtures: from synthetic cilia to flowing networks

    NASA Astrophysics Data System (ADS)

    Sanchez, Tim

    2013-03-01

    Inspired by biological functions such as ciliary beating and cytoplasmic streaming, we have developed a highly tunable and robust model system from biological components that self-organizes to produce a broad range of far-from-equilibrium materials with remarkable emergent properties. Using only simple components - microtubules, kinesin motor clusters, and a depletion agent that bundles MTs - we reproduced several essential biological functions, including cilia-like beating, the emergence of metachronal waves in bundle arrays, and internally generated flows in active cytoskeletal gels. The occurrence of these biomimetic functions as self-organized processes provides unique insight into the mechanisms that drive these processes in biology. Beyond these biomimetic behaviors, we have also used the same components to engineer novel active materials which have no biological analogues: active streaming 2D nematics, and finally self-propelled emulsion droplets. These observations exemplify how assemblages of animate microscopic objects exhibit highly sought-after collective and biomimetic properties, challenging us to develop a theoretical framework that would allow for a systematic engineering of their far-from-equilibrium material properties.

  17. PEX6 is Expressed in Photoreceptor Cilia and Mutated in Deafblindness with Enamel Dysplasia and Microcephaly.

    PubMed

    Zaki, Maha S; Heller, Raoul; Thoenes, Michaela; Nürnberg, Gudrun; Stern-Schneider, Gabi; Nürnberg, Peter; Karnati, Srikanth; Swan, Daniel; Fateen, Ekram; Nagel-Wolfrum, Kerstin; Mostafa, Mostafa I; Thiele, Holger; Wolfrum, Uwe; Baumgart-Vogt, Eveline; Bolz, Hanno J

    2016-02-01

    Deafblindness is part of several genetic disorders. We investigated a consanguineous Egyptian family with two siblings affected by congenital hearing loss and retinal degeneration, initially diagnosed as Usher syndrome type 1. At teenage, severe enamel dysplasia, developmental delay, and microcephaly became apparent. Genome-wide homozygosity mapping and whole-exome sequencing detected a homozygous missense mutation, c.1238G>T (p.Gly413Val), affecting a highly conserved residue of peroxisomal biogenesis factor 6, PEX6. Biochemical profiling of the siblings revealed abnormal and borderline plasma phytanic acid concentration, and cerebral imaging revealed white matter disease in both. We show that Pex6 localizes to the apical extensions of secretory ameloblasts and differentiated odontoblasts at early stages of dentin synthesis in mice, and to cilia of retinal photoreceptor cells. We propose PEX6, and possibly other peroxisomal genes, as candidate for the rare cooccurrence of deafblindness and enamel dysplasia. Our study for the first time links peroxisome biogenesis disorders to retinal ciliopathies. PMID:26593283

  18. The effect of Gd on trityl-based dynamic nuclear polarisation in solids.

    PubMed

    Ravera, Enrico; Shimon, Daphna; Feintuch, Akiva; Goldfarb, Daniella; Vega, Shimon; Flori, Alessandra; Luchinat, Claudio; Menichetti, Luca; Parigi, Giacomo

    2015-10-28

    In dynamic nuclear polarisation (DNP) experiments performed under static conditions at 1.4 K we show that the presence of 1 mM Gd(iii)-DOTAREM increases the (13)C polarisation and decreases the (13)C polarisation buildup time of (13)C-urea dissolved in samples containing water/DMSO mixtures with trityl radical (OX063) concentrations of 10 mM or higher. To account for these observations further measurements were carried out at 6.5 K, using a combined EPR and NMR spectrometer. At this temperature, frequency swept DNP spectra of samples with 5 or 10 mM OX063 were measured, with and without 1 mM Gd-DOTA, and again a (13)C enhancement gain was observed due to the presence of Gd-DOTA. These measurements were complemented by electron-electron double resonance (ELDOR) measurements to quantitate the effect of electron spectral diffusion (eSD) on the DNP enhancements and lineshapes. Simulations of the ELDOR spectra were done using the following parameters: (i) a parameter defining the rate of the eSD process, (ii) an "effective electron-proton anisotropic hyperfine interaction parameter", and (iii) the transverse electron spin relaxation time of OX063. These parameters, together with the longitudinal electron spin relaxation time, measured by EPR, were used to calculate the frequency profile of electron polarisation. This, in turn, was used to calculate two basic solid effect (SE) and indirect cross effect (iCE) DNP spectra. A properly weighted combination of these two normalized DNP spectra provided a very good fit of the experimental DNP spectra. The best fit simulation parameters reveal that the addition of Gd(iii)-DOTA causes an increase in both the SE and the iCE contributions by similar amounts, and that the increase in the overall DNP enhancements is a result of narrowing of the ELDOR spectra (increased electron polarisation gradient across the EPR line). These changes in the electron depolarisation profile are a combined result of shortening of the longitudinal and

  19. QUIJOTE scientific results - I. Measurements of the intensity and polarisation of the anomalous microwave emission in the Perseus molecular complex

    NASA Astrophysics Data System (ADS)

    Génova-Santos, R.; Rubiño-Martín, J. A.; Rebolo, R.; Peláez-Santos, A.; López-Caraballo, C. H.; Harper, S.; Watson, R. A.; Ashdown, M.; Barreiro, R. B.; Casaponsa, B.; Dickinson, C.; Diego, J. M.; Fernández-Cobos, R.; Grainge, K. J. B.; Gutiérrez, C. M.; Herranz, D.; Hoyland, R.; Lasenby, A.; López-Caniego, M.; Martínez-González, E.; McCulloch, M.; Melhuish, S.; Piccirillo, L.; Perrott, Y. C.; Poidevin, F.; Razavi-Ghods, N.; Scott, P. F.; Titterington, D.; Tramonte, D.; Vielva, P.; Vignaga, R.

    2015-10-01

    In this paper, we present Q-U-I JOint Tenerife Experiment (QUIJOTE) 10-20 GHz observations (194 h in total over ≈250 deg2) in intensity and polarisation of G159.6-18.5, one of the most widely studied regions harbouring anomalous microwave emission (AME). By combining with other publicly available intensity data, we achieve the most precise spectrum of the AME measured to date in an individual region, with 13 independent data points between 10 and 50 GHz being dominated by this emission. The four QUIJOTE data points provide the first independent confirmation of the downturn of the AME spectrum at low frequencies, initially unveiled by the COSMOlogical Structures On Medium Angular Scales experiment in this region. Our polarisation maps, which have an angular resolution of ≈1° and a sensitivity of ≈ 25 μK beam-1, are consistent with zero polarisation. We obtain upper limits on the polarisation fraction of Π < 6.3 and <2.8 per cent (95 per cent C.L.), respectively, at 12 and 18 GHz (ΠAME < 10.1 and <3.4 per cent with respect to the residual AME intensity), a frequency range where no AME polarisation observations have been reported to date. The combination of these constraints with those from other experiments confirm that all the magnetic dust models based on single-domain grains, and most of those considering randomly oriented magnetic inclusions, predict higher polarisation levels than is observed towards regions with AME. Also, neither of the two considered models of electric dipole emission seems to be compatible with all the observations together. More stringent constraints of the AME polarisation at 10-40 GHz are necessary to disentangle between different models, to which future QUIJOTE data will contribute.

  20. Recent advances in primary ciliary dyskinesia.

    PubMed

    Takeuchi, Kazuhiko; Kitano, Masako; Ishinaga, Hajime; Kobayashi, Masayoshi; Ogawa, Satoru; Nakatani, Kaname; Masuda, Sawako; Nagao, Mizuho; Fujisawa, Takao

    2016-06-01

    Primary ciliary dyskinesia (PCD) is a genetic disease inherited in an autosomal recessive manner. The prevalence of PCD is estimated to be 1 in 20,000 live births. Congenital abnormality of the primary cilia results in situs inversus in 50% of patients. Decreased function of motile cilia causes chronic rhinosinusitis, otitis media with effusion, bronchiectasis and infertility. Cases with situs inversus are considered to show "Kartagener's syndrome", and diagnosis is not difficult. However, in cases without situs inversus, the diagnosis is much more troublesome. PCD without situs inversus is thus probably underdiagnosed. Prolonged chronic cough represents an important symptom that is seen in most patients. The diagnosis of PCD requires the presence of the characteristic clinical phenotypes and either: (1) specific ciliary ultrastructural defects identified by transmission electron microscopy in biopsy samples of respiratory epithelium; or (2) identification of mutation in one of the genes known to be associated with PCD. Nasal nitric oxide concentration is extremely low in PCD, and this could be useful for screening of the disease. At present, no fundamental therapies are available for PCD. Diagnosis in the early stages is important to prevent progression of bronchiectasis and deterioration of lung function by guidance for daily life, immunization, cessation of smoking and prompt therapy at the time of respiratory tract infection. Since PCD is inherited in an autosomal-recessive manner, genetic counseling is necessary after definite diagnosis. PMID:26527516

  1. Polarisation dependence of Schottky barrier heights at ferroelectric BaTiO3 / RuO2 interfaces: influence of substrate orientation and quality

    NASA Astrophysics Data System (ADS)

    Hubmann, Andreas H.; Li, Shunyi; Zhukov, Sergey; von Seggern, Heinz; Klein, Andreas

    2016-07-01

    In situ x-ray photoelectron spectroscopy was employed to examine the change in Schottky barrier height Δ {Φ\\text{B}} at BaTiO3/RuO2 interfaces upon polarisation reversal for [1 0 0], [1 1 0] and [1 1 1] oriented BaTiO3 single crystals. Compared to previous measurements on BaTiO3/RuO2 interfaces (Chen and Klein 2012 Phys. Rev. B 86 094105), the crystals exhibit a significantly reduced dependence of barrier height on polarisation direction. This is connected to a much higher polarisation of the present cystals, which is comparable to the accepted bulk polarisation of BaTiO3 of 26~μ \\text{C}~\\text{c}{{\\text{m}}-2} and which exhibit the expected dependence on crystal orientation. This indicates a much higher crystal quality in the present experiments, which is also confirmed by a Kolmogorov–Avrami–Ishibashi like polarisation switching dynamics. It is observed that Δ {Φ\\text{B}} is reduced for the [1 1 0] and [1 1 1] orientation and scales with polarisation as long as crystals from the same batch are used. The fact, that a poor polarisation hysteresis behaviour relates to a high polarisation dependence of Schottky barrier height, indicates that the electrode’s ability to screen ferroelectric polarisation charges depends sensitively on crystal and/or interface quality.

  2. SPECIAL ISSUE DEVOTED TO THE 80TH ANNIVERSARY OF ACADEMICIAN N G BASOV'S BIRTH: An optically polarised dense 3He target as a spin filter for slow-neutron beams

    NASA Astrophysics Data System (ADS)

    Kolachevsky, Nikolai N.; Prokof'ichev, Yu V.; Skoi, V. R.; Sobel'man, Igor I.; Sorokin, Vadim N.

    2003-01-01

    The possibility of polarising 3He and preserving the polarisation in an external magnetic field of 0.05 Oe is demonstrated experimentally. A neutron filter with an extremely weak guiding field is fabricated for obtaining polarised neutron beams. The degree of polarisation equal to 25% was obtained for 0.025-eV neutrons. Some fields of application of other polarised noble gases are considered. The cross section for cross-relaxation of nuclear polarisation in the 129Xe — 131Xe mixture is estimated.

  3. Why motor proteins team up - Intraflagellar transport in C. elegans cilia

    PubMed Central

    Mijalkovic, Jona; Prevo, Bram; Peterman, Erwin J. G.

    2016-01-01

    ABSTRACT Inside the cell, vital processes such as cell division and intracellular transport are driven by the concerted action of different molecular motor proteins. In C. elegans chemosensory cilia, 2 kinesin-2 family motor proteins, kinesin-II and OSM-3, team up to drive intraflagellar transport (IFT) in the anterograde direction, from base to tip, whereas IFT dynein hitchhikes toward the tip and subsequently drives IFT in the opposite, retrograde direction, thereby recycling both kinesins. While it is evident that at least a retrograde and an anterograde motor are necessary to drive IFT, it has remained puzzling why 2 same-polarity kinesins are employed. Recently, we addressed this question by combining advanced genome-engineering tools with ultrasensitive, quantitative fluorescence microscopy to study IFT with single-molecule sensitivity.1,2 Using this combination of approaches, we uncovered a differentiation in kinesin-2 function, in which the slower kinesin-II operates as an ‘importer’, loading IFT trains into the cilium before gradually handing them over to the faster OSM-3. OSM-3 subsequently acts as a long-range ‘transporter’, driving the IFT trains toward the tip. The two kinesin-2 motors combine their unique motility properties to achieve something neither motor can achieve on its own; that is to optimize the amount of cargo inside the cilium. In this commentary, we provide detailed insight into the rationale behind our research approach and comment on our recent findings. Moreover, we discuss the role of IFT dynein and provide an outlook on future studies. PMID:27384150

  4. Dexmedetomidine and Fentanyl Exhibit Temperature Dependent Effects on Human Respiratory Cilia

    PubMed Central

    Welchering, Nils; Ochoa, Sebastian; Tian, Xin; Francis, Richard; Zahid, Maliha; Muñoz, Ricardo; Lo, Cecilia W.

    2015-01-01

    Background: Dexmedetomidine (dex) is commonly used in intensive care due to its effective sedation and analgesia with few adverse effects and minimal respiratory depression. However, we recently observed that exposing mouse epithelial respiratory cells to dex decreased ciliary beat frequency (CBF), suggesting dex may pose pulmonary risk. Objective: The purpose of this study is to determine the effects of dex at clinically relevant doses on CBF in human respiratory epithelia. Methods: Human nasal epithelial cilia were obtained from the inferior nasal turbinate with a rhinoprobe and placed in culture medium at 15°C and 37°C. At 5 and 30 min, video-microscopy was used to assess CBF, either without (control) or with different concentrations (1, 5, and 10 nM) of dex, fentanyl (fen), and dex + fen combination. Results: At 15°C, CBF was lower in the dex group compared to controls at 5 and 30 min. At 37°C, there was a significant increase in CBF with dex at 5 and 30 min, except for dex at 5 nM after 5 min, which showed a significant decrease. At 15°C the combination of dex + fen showed a positive interaction, causing less ciliary inhibition as expected. In contrast, no interaction between drugs was seen between dex and fen at 37°C. Conclusion: At low temperatures, dex reduces CBF in human respiratory epithelia, whereas dex increases CBF at physiologic temperature in vitro. Whether these effects translate into clinical consequences during hypothermia, as with cardiopulmonary bypass surgery will require further studies. PMID:25717467

  5. Late summer sea ice segmentation with multi-polarisation SAR features in C- and X-band

    NASA Astrophysics Data System (ADS)

    Fors, A. S.; Brekke, C.; Doulgeris, A. P.; Eltoft, T.; Renner, A. H. H.; Gerland, S.

    2015-09-01

    In this study we investigate the potential of sea ice segmentation by C- and X-band multi-polarisation synthetic aperture radar (SAR) features during late summer. Five high-resolution satellite SAR scenes were recorded in the Fram Strait covering iceberg-fast first-year and old sea ice during a week with air temperatures varying around zero degrees Celsius. In situ data consisting of sea ice thickness, surface roughness and aerial photographs were collected during a helicopter flight at the site. Six polarimetric SAR features were extracted for each of the scenes. The ability of the individual SAR features to discriminate between sea ice types and their temporally consistency were examined. All SAR features were found to add value to sea ice type discrimination. Relative kurtosis, geometric brightness, cross-polarisation ratio and co-polarisation correlation angle were found to be temporally consistent in the investigated period, while co-polarisation ratio and co-polarisation correlation magnitude were found to be temporally inconsistent. An automatic feature-based segmentation algorithm was tested both for a full SAR feature set, and for a reduced SAR feature set limited to temporally consistent features. In general, the algorithm produces a good late summer sea ice segmentation. Excluding temporally inconsistent SAR features improved the segmentation at air temperatures above zero degrees Celcius.

  6. The Mitotic Exit Network and Cdc14 phosphatase initiate cytokinesis by counteracting CDK phosphorylations and blocking polarised growth

    PubMed Central

    Sanchez-Diaz, Alberto; Nkosi, Pedro Junior; Murray, Stephen; Labib, Karim

    2012-01-01

    Polarisation of the actin cytoskeleton must cease during cytokinesis, to support efficient assembly and contraction of the actomyosin ring at the site of cell division, but the underlying mechanisms are still understood poorly in most species. In budding yeast, the Mitotic Exit Network (MEN) releases Cdc14 phosphatase from the nucleolus during anaphase, leading to the inactivation of mitotic forms of cyclin-dependent kinase (CDK) and the onset of septation, before G1-CDK can be reactivated and drive re-polarisation of the actin cytoskeleton to a new bud. Here, we show that premature inactivation of mitotic CDK, before release of Cdc14, allows G1-CDK to divert the actin cytoskeleton away from the actomyosin ring to a new site of polarised growth, thereby delaying progression through cytokinesis. Our data indicate that cells normally avoid this problem via the MEN-dependent release of Cdc14, which counteracts all classes of CDK-mediated phosphorylations during cytokinesis and blocks polarised growth. The dephosphorylation of CDK targets is therefore central to the mechanism by which the MEN and Cdc14 initiate cytokinesis and block polarised growth during late mitosis. PMID:22872148

  7. Late-summer sea ice segmentation with multi-polarisation SAR features in C and X band

    NASA Astrophysics Data System (ADS)

    Fors, Ane S.; Brekke, Camilla; Doulgeris, Anthony P.; Eltoft, Torbjørn; Renner, Angelika H. H.; Gerland, Sebastian

    2016-02-01

    In this study, we investigate the potential of sea ice segmentation by C- and X-band multi-polarisation synthetic aperture radar (SAR) features during late summer. Five high-resolution satellite SAR scenes were recorded in the Fram Strait covering iceberg-fast first-year and old sea ice during a week with air temperatures varying around 0 °C. Sea ice thickness, surface roughness and aerial photographs were collected during a helicopter flight at the site. Six polarimetric SAR features were extracted for each of the scenes. The ability of the individual SAR features to discriminate between sea ice types and their temporal consistency were examined. All SAR features were found to add value to sea ice type discrimination. Relative kurtosis, geometric brightness, cross-polarisation ratio and co-polarisation correlation angle were found to be temporally consistent in the investigated period, while co-polarisation ratio and co-polarisation correlation magnitude were found to be temporally inconsistent. An automatic feature-based segmentation algorithm was tested both for a full SAR feature set and for a reduced SAR feature set limited to temporally consistent features. In C band, the algorithm produced a good late-summer sea ice segmentation, separating the scenes into segments that could be associated with different sea ice types in the next step. The X-band performance was slightly poorer. Excluding temporally inconsistent SAR features improved the segmentation in one of the X-band scenes.

  8. Primary Ciliary Dyskinesia

    PubMed Central

    Lobo, Jason; Zariwala, Maimoona A; Noone, Peadar G

    2016-01-01

    Primary ciliary dyskinesia (PCD) is an autosomal recessive disorder of cilia structure, function, and biogenesis leading to chronic infections of the respiratory tract, fertility problems and disorders of organ laterality. The diagnosis can be challenging, using traditional tools such as characteristic clinical features, ciliary functional and ultra-structural defects; newer screening tools such as nasal nitric oxide levels and genetic testing add to the diagnostic algorithm. There are thirty-two known PCD causing genes, and in the future, comprehensive genetic testing may screen young infants prior to developing symptoms thus improving survival. Therapies include surveillance of pulmonary function and microbiology, in addition to airway clearance, antibiotics and ideally, early referral to bronchiectasis centers. As with CF, standardized care at specialized centers using a multidisciplinary approach likely improves outcomes. In conjunction with the CF foundation, the PCD foundation, and with lead investigators and clinicians, is developing a network of PCD clinical centers to coordinate the effort in North America and Europe. As the network grows, care and knowledge will improve. PMID:25826585

  9. Primary ciliary dyskinesia.

    PubMed

    Lobo, Jason; Zariwala, Maimoona A; Noone, Peadar G

    2015-04-01

    Primary ciliary dyskinesia (PCD) is an autosomal recessive disorder of cilia structure, function, and biogenesis leading to chronic infections of the respiratory tract, fertility problems, and disorders of organ laterality. The diagnosis can be challenging, using traditional tools such as characteristic clinical features, ciliary function, and ultrastructural defects and newer screening tools such as nasal nitric oxide levels and genetic testing add to the diagnostic algorithm. There are 32 known PCD-causing genes, and in the future, comprehensive genetic testing may screen young infants before developing symptoms, thus improving survival. Therapies include surveillance of pulmonary function and microbiology, in addition to airway clearance, antibiotics, and early referral to bronchiectasis centers. As with cystic fibrosis (CF), standardized care at specialized centers using a multidisciplinary approach likely improves outcomes. In conjunction with the CF foundation, the PCD foundation, with experienced investigators and clinicians, is developing a network of PCD clinical centers to coordinate the effort in North America and Europe. As the network grows, clinical care and knowledge will improve. PMID:25826585

  10. Chitosan drives anti-inflammatory macrophage polarisation and pro-inflammatory dendritic cell stimulation.

    PubMed

    Oliveira, Marta I; Santos, Susana G; Oliveira, Maria J; Torres, Ana L; Barbosa, Mário A

    2012-01-01

    Macrophages and dendritic cells (DC) share the same precursor and play key roles in immunity. Modulation of their behaviour to achieve an optimal host response towards an implanted device is still a challenge. Here we compare the differentiation process and polarisation of these related cell populations and show that they exhibit different responses to chitosan (Ch), with human monocyte-derived macrophages polarising towards an anti-inflammatory phenotype while their DC counterparts display pro-inflammatory features. Macrophages and DC, whose interactions with biomaterials are frequently analysed using fully differentiated cells, were cultured directly on Ch films, rather than exposed to the polymer after complete differentiation. Ch was the sole stimulating factor and activated both macrophages and DC, without leading to significant T cell proliferation. After 10 d on Ch, macrophages significantly down-regulated expression of pro-inflammatory markers, CD86 and MHCII. Production of pro-inflammatory cytokines, particularly TNF-α, decreased with time for cells cultured on Ch, while anti-inflammatory IL-10 and TGF-β1, significantly increased. Altogether, these results suggest an M2c polarisation. Also, macrophage matrix metalloproteinase activity was augmented and cell motility was stimulated by Ch. Conversely, DC significantly enhanced CD86 expression, reduced IL-10 secretion and increased TNF-α and IL-1β levels. Our findings indicate that cells with a common precursor may display different responses, when challenged by the same biomaterial. Moreover, they help to further comprehend macrophage/DC interactions with Ch and the balance between pro- and anti-inflammatory signals associated with implant biomaterials. We propose that an overall pro-inflammatory reaction may hide the expression of anti-inflammatory cytokines, likely relevant for tissue repair/regeneration. PMID:22828991

  11. Spatiotemporal Cadence of Macrophage Polarisation in a Model of Light-Induced Retinal Degeneration

    PubMed Central

    Valter, Krisztina; Provis, Jan M.; Rutar, Matt

    2015-01-01

    Background The recruitment of macrophages accompanies almost every pathogenic state of the retina, and their excessive activation in the subretinal space is thought to contribute to the progression of diseases including age-related macular degeneration. Previously, we have shown that macrophages aggregate in the outer retina following damage elicited by photo-oxidative stress, and that inhibition of their recruitment reduces photoreceptor death. Here, we look for functional insight into macrophage activity in this model through the spatiotemporal interplay of macrophage polarisation over the course of degeneration. Methods Rats were exposed to 1000 lux light damage (LD) for 24hrs, with some left to recover for 3 and 7 days post-exposure. Expression and localisation of M1- and M2- macrophage markers was investigated in light-damaged retinas using qPCR, ELISA, flow cytometry, and immunohistochemistry. Results Expression of M1- (Ccl3, Il-6, Il-12, Il-1β, TNFα) and M2- (CD206, Arg1, Igf1, Lyve1, Clec7a) related markers followed discrete profiles following light damage; up-regulation of M1 genes peaked at the early phase of cell death, while M2 genes generally exhibited more prolonged increases during the chronic phase. Moreover, Il-1β and CD206 labelled accumulations of microglia/macrophages which differed in their morphological, temporal, and spatial characteristics following light damage. Conclusions The data illustrate a dynamic shift in macrophage polarisation following light damage through a broad swathe of M1 and M2 markers. Pro-inflammatory M1 activation appears to dominate the early phase of degeneration while M2 responses appear to more heavily mark the chronic post-exposure period. While M1/M2 polarisation represents two extremes amongst a spectrum of macrophage activity, knowledge of their predominance offers insight into functional consequences of macrophage activity over the course of damage, which may inform the spatiotemporal employment of

  12. Whole exome re-sequencing implicates CCDC38 and cilia structure and function in resistance to smoking related airflow obstruction.

    PubMed

    Wain, Louise V; Sayers, Ian; Soler Artigas, María; Portelli, Michael A; Zeggini, Eleftheria; Obeidat, Ma'en; Sin, Don D; Bossé, Yohan; Nickle, David; Brandsma, Corry-Anke; Malarstig, Anders; Vangjeli, Ciara; Jelinsky, Scott A; John, Sally; Kilty, Iain; McKeever, Tricia; Shrine, Nick R G; Cook, James P; Patel, Shrina; Spector, Tim D; Hollox, Edward J; Hall, Ian P; Tobin, Martin D

    2014-05-01

    Chronic obstructive pulmonary disease (COPD) is a leading cause of global morbidity and mortality and, whilst smoking remains the single most important risk factor, COPD risk is heritable. Of 26 independent genomic regions showing association with lung function in genome-wide association studies, eleven have been reported to show association with airflow obstruction. Although the main risk factor for COPD is smoking, some individuals are observed to have a high forced expired volume in 1 second (FEV1) despite many years of heavy smoking. We hypothesised that these "resistant smokers" may harbour variants which protect against lung function decline caused by smoking and provide insight into the genetic determinants of lung health. We undertook whole exome re-sequencing of 100 heavy smokers who had healthy lung function given their age, sex, height and smoking history and applied three complementary approaches to explore the genetic architecture of smoking resistance. Firstly, we identified novel functional variants in the "resistant smokers" and looked for enrichment of these novel variants within biological pathways. Secondly, we undertook association testing of all exonic variants individually with two independent control sets. Thirdly, we undertook gene-based association testing of all exonic variants. Our strongest signal of association with smoking resistance for a non-synonymous SNP was for rs10859974 (P = 2.34 × 10(-4)) in CCDC38, a gene which has previously been reported to show association with FEV1/FVC, and we demonstrate moderate expression of CCDC38 in bronchial epithelial cells. We identified an enrichment of novel putatively functional variants in genes related to cilia structure and function in resistant smokers. Ciliary function abnormalities are known to be associated with both smoking and reduced mucociliary clearance in patients with COPD. We suggest that genetic influences on the development or function of cilia in the bronchial epithelium may

  13. Studies on cilia. 3. Further studies on the cilium tip and a "sliding filament" model of ciliary motility.