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Sample records for drosophila segment determination

  1. Drosophila Hox and Sex-Determination Genes Control Segment Elimination through EGFR and extramacrochetae Activity

    PubMed Central

    Foronda, David; Martín, Paloma; Sánchez-Herrero, Ernesto

    2012-01-01

    The formation or suppression of particular structures is a major change occurring in development and evolution. One example of such change is the absence of the seventh abdominal segment (A7) in Drosophila males. We show here that there is a down-regulation of EGFR activity and fewer histoblasts in the male A7 in early pupae. If this activity is elevated, cell number increases and a small segment develops in the adult. At later pupal stages, the remaining precursors of the A7 are extruded under the epithelium. This extrusion requires the up-regulation of the HLH protein Extramacrochetae and correlates with high levels of spaghetti-squash, the gene encoding the regulatory light chain of the non-muscle myosin II. The Hox gene Abdominal-B controls both the down-regulation of spitz, a ligand of the EGFR pathway, and the up-regulation of extramacrochetae, and also regulates the transcription of the sex-determining gene doublesex. The male Doublesex protein, in turn, controls extramacrochetae and spaghetti-squash expression. In females, the EGFR pathway is also down-regulated in the A7 but extramacrochetae and spaghetti-squash are not up-regulated and extrusion of precursor cells is almost absent. Our results show the complex orchestration of cellular and genetic events that lead to this important sexually dimorphic character change. PMID:22912593

  2. Drosophila Hox and sex-determination genes control segment elimination through EGFR and extramacrochetae activity.

    PubMed

    Foronda, David; Martín, Paloma; Sánchez-Herrero, Ernesto

    2012-01-01

    The formation or suppression of particular structures is a major change occurring in development and evolution. One example of such change is the absence of the seventh abdominal segment (A7) in Drosophila males. We show here that there is a down-regulation of EGFR activity and fewer histoblasts in the male A7 in early pupae. If this activity is elevated, cell number increases and a small segment develops in the adult. At later pupal stages, the remaining precursors of the A7 are extruded under the epithelium. This extrusion requires the up-regulation of the HLH protein Extramacrochetae and correlates with high levels of spaghetti-squash, the gene encoding the regulatory light chain of the non-muscle myosin II. The Hox gene Abdominal-B controls both the down-regulation of spitz, a ligand of the EGFR pathway, and the up-regulation of extramacrochetae, and also regulates the transcription of the sex-determining gene doublesex. The male Doublesex protein, in turn, controls extramacrochetae and spaghetti-squash expression. In females, the EGFR pathway is also down-regulated in the A7 but extramacrochetae and spaghetti-squash are not up-regulated and extrusion of precursor cells is almost absent. Our results show the complex orchestration of cellular and genetic events that lead to this important sexually dimorphic character change. PMID:22912593

  3. Generating patterns from fields of cells. Examples from Drosophila segmentation.

    PubMed

    Sanson, B

    2001-12-01

    In Drosophila, a cascade of maternal, gap, pair-rule and segment polarity genes subdivides the antero/posterior axis of the embryo into repeating segmental stripes. This review summarizes what happens next, i.e. how an intrasegmental pattern is generated and controls the differentiation of specific cell types in the epidermis. Within each segment, cells secreting the signalling molecules Wingless (the homologue of vertebrate Wnt-1) and Hedgehog are found in narrow stripes on both sides of the parasegmental boundary. The Wingless and Hedgehog organizing activities help to establish two more stripes per segment that localize ligands for the Epidermal Growth Factor and the Notch signalling pathways, respectively. These four signals then act at short range and in concert to control epidermal differentiation at the single cell level across the segment. This example from Drosophila provides a paradigm for how organizers generate precise patterns, and ultimately different cell types, in a naïve field of cells. PMID:11743020

  4. Topology and Robustness in the Drosophila Segment Polarity Network

    PubMed Central

    2004-01-01

    A complex hierarchy of genetic interactions converts a single-celled Drosophila melanogaster egg into a multicellular embryo with 14 segments. Previously, von Dassow et al. reported that a mathematical model of the genetic interactions that defined the polarity of segments (the segment polarity network) was robust (von Dassow et al. 2000). As quantitative information about the system was unavailable, parameters were sampled randomly. A surprisingly large fraction of these parameter sets allowed the model to maintain and elaborate on the segment polarity pattern. This robustness is due to the positive feedback of gene products on their own expression, which induces individual cells in a model segment to adopt different stable expression states (bistability) corresponding to different cell types in the segment polarity pattern. A positive feedback loop will only yield multiple stable states when the parameters that describe it satisfy a particular inequality. By testing which random parameter sets satisfy these inequalities, I show that bistability is necessary to form the segment polarity pattern and serves as a strong predictor of which parameter sets will succeed in forming the pattern. Although the original model was robust to parameter variation, it could not reproduce the observed effects of cell division on the pattern of gene expression. I present a modified version that incorporates recent experimental evidence and does successfully mimic the consequences of cell division. The behavior of this modified model can also be understood in terms of bistability in positive feedback of gene expression. I discuss how this topological property of networks provides robust pattern formation and how large changes in parameters can change the specific pattern produced by a network. PMID:15208707

  5. Transcriptional control in the segmentation gene network of Drosophila.

    PubMed

    Schroeder, Mark D; Pearce, Michael; Fak, John; Fan, HongQing; Unnerstall, Ulrich; Emberly, Eldon; Rajewsky, Nikolaus; Siggia, Eric D; Gaul, Ulrike

    2004-09-01

    The segmentation gene network of Drosophila consists of maternal and zygotic factors that generate, by transcriptional (cross-) regulation, expression patterns of increasing complexity along the anterior-posterior axis of the embryo. Using known binding site information for maternal and zygotic gap transcription factors, the computer algorithm Ahab recovers known segmentation control elements (modules) with excellent success and predicts many novel modules within the network and genome-wide. We show that novel module predictions are highly enriched in the network and typically clustered proximal to the promoter, not only upstream, but also in intronic space and downstream. When placed upstream of a reporter gene, they consistently drive patterned blastoderm expression, in most cases faithfully producing one or more pattern elements of the endogenous gene. Moreover, we demonstrate for the entire set of known and newly validated modules that Ahab's prediction of binding sites correlates well with the expression patterns produced by the modules, revealing basic rules governing their composition. Specifically, we show that maternal factors consistently act as activators and that gap factors act as repressors, except for the bimodal factor Hunchback. Our data suggest a simple context-dependent rule for its switch from repressive to activating function. Overall, the composition of modules appears well fitted to the spatiotemporal distribution of their positive and negative input factors. Finally, by comparing Ahab predictions with different categories of transcription factor input, we confirm the global regulatory structure of the segmentation gene network, but find odd skipped behaving like a primary pair-rule gene. The study expands our knowledge of the segmentation gene network by increasing the number of experimentally tested modules by 50%. For the first time, the entire set of validated modules is analyzed for binding site composition under a uniform set of

  6. Transcriptional Control in the Segmentation Gene Network of Drosophila

    PubMed Central

    Fan, HongQing; Unnerstall, Ulrich; Emberly, Eldon; Rajewsky, Nikolaus; Siggia, Eric D

    2004-01-01

    The segmentation gene network of Drosophila consists of maternal and zygotic factors that generate, by transcriptional (cross-) regulation, expression patterns of increasing complexity along the anterior-posterior axis of the embryo. Using known binding site information for maternal and zygotic gap transcription factors, the computer algorithm Ahab recovers known segmentation control elements (modules) with excellent success and predicts many novel modules within the network and genome-wide. We show that novel module predictions are highly enriched in the network and typically clustered proximal to the promoter, not only upstream, but also in intronic space and downstream. When placed upstream of a reporter gene, they consistently drive patterned blastoderm expression, in most cases faithfully producing one or more pattern elements of the endogenous gene. Moreover, we demonstrate for the entire set of known and newly validated modules that Ahab's prediction of binding sites correlates well with the expression patterns produced by the modules, revealing basic rules governing their composition. Specifically, we show that maternal factors consistently act as activators and that gap factors act as repressors, except for the bimodal factor Hunchback. Our data suggest a simple context-dependent rule for its switch from repressive to activating function. Overall, the composition of modules appears well fitted to the spatiotemporal distribution of their positive and negative input factors. Finally, by comparing Ahab predictions with different categories of transcription factor input, we confirm the global regulatory structure of the segmentation gene network, but find odd skipped behaving like a primary pair-rule gene. The study expands our knowledge of the segmentation gene network by increasing the number of experimentally tested modules by 50%. For the first time, the entire set of validated modules is analyzed for binding site composition under a uniform set of

  7. Captured segment exchange: a strategy for custom engineering large genomic regions in Drosophila melanogaster.

    PubMed

    Bateman, Jack R; Palopoli, Michael F; Dale, Sarah T; Stauffer, Jennifer E; Shah, Anita L; Johnson, Justine E; Walsh, Conor W; Flaten, Hanna; Parsons, Christine M

    2013-02-01

    Site-specific recombinases (SSRs) are valuable tools for manipulating genomes. In Drosophila, thousands of transgenic insertions carrying SSR recognition sites have been distributed throughout the genome by several large-scale projects. Here we describe a method with the potential to use these insertions to make custom alterations to the Drosophila genome in vivo. Specifically, by employing recombineering techniques and a dual recombinase-mediated cassette exchange strategy based on the phiC31 integrase and FLP recombinase, we show that a large genomic segment that lies between two SSR recognition-site insertions can be "captured" as a target cassette and exchanged for a sequence that was engineered in bacterial cells. We demonstrate this approach by targeting a 50-kb segment spanning the tsh gene, replacing the existing segment with corresponding recombineered sequences through simple and efficient manipulations. Given the high density of SSR recognition-site insertions in Drosophila, our method affords a straightforward and highly efficient approach to explore gene function in situ for a substantial portion of the Drosophila genome. PMID:23150604

  8. How Notch establishes longitudinal axon connections between successive segments of the Drosophila CNS.

    PubMed

    Kuzina, Irina; Song, Jeong K; Giniger, Edward

    2011-05-01

    Development of the segmented central nerve cords of vertebrates and invertebrates requires connecting successive neuromeres. Here, we show both how a pathway is constructed to guide pioneer axons between segments of the Drosophila CNS, and how motility of the pioneers along that pathway is promoted. First, canonical Notch signaling in specialized glial cells causes nearby differentiating neurons to extrude a mesh of fine projections, and shapes that mesh into a continuous carpet that bridges from segment to segment, hugging the glial surface. This is the direct substratum that pioneer axons follow as they grow. Simultaneously, Notch uses an alternate, non-canonical signaling pathway in the pioneer growth cones themselves, promoting their motility by suppressing Abl signaling to stimulate filopodial growth while presumably reducing substratum adhesion. This propels the axons as they establish the connection between successive segments. PMID:21447553

  9. Laser ablation of persistent twist cells in Drosophila: muscle precursor fate is not segmentally restricted

    NASA Technical Reports Server (NTRS)

    Farrell, E. R.; Keshishian, H.

    1999-01-01

    In Drosophila the precursors of the adult musculature arise during embryogenesis. These precursor cells have been termed Persistent Twist Cells (PTCs), as they continue to express the transcription factor Twist after that gene ceases expression elsewhere in the mesoderm. In the larval abdomen, the PTCs are associated with peripheral nerves in stereotypic ventral, dorsal, and lateral clusters, which give rise, respectively, to the ventral, dorsal, and lateral muscle fiber groups of the adult. We tested the developmental potential of the PTCs by using a microbeam laser to ablate specific clusters in larvae. We found that the ablation of a single segmental PTC cluster does not usually result in the deletion of the corresponding adult fibers of that segment. Instead, normal or near normal numbers of adult fibers can form after the ablation. Examination of pupae following ablation showed that migrating PTCs from adjacent segments are able to invade the affected segment, replenishing the ablated cells. However, the ablation of homologous PTCs in multiple segments does result in the deletion of the corresponding adult muscle fibers. These data indicate that the PTCs in an abdominal segment can contribute to the formation of muscle fibers in adjacent abdominal segments, and thus are not inherently restricted to the formation of muscle fibers within their segment of origin.

  10. Haplotype test reveals departure from neutrality in a segment of the white gene of Drosophila melanogaster

    SciTech Connect

    Kirby, D.A.; Stephan, W.

    1995-12-01

    Restriction map studies previously revealed extensive linkage disequilibria in the transcriptional unit of the white locus in natural Drosophila melanogaster populations. To understand the causes of these disequilibria, we sequenced a 4722-bp region of the white gene from 15 lines of D. melanogaster and 1 line of Drosophila simulans. Statistical tests applied to the entire 4722-bp region do not reject neutrality. In contrast, a test for high-frequency haplotypes ({open_quotes}Haplotype test{close_quotes}) revealed an 834-bp segment, encompassing the 3{prime} end of intron 1 to the 3{prime} end of intron 2, in which the structure of variation deviates significantly from the predictions of a neutral equilibrium model. The variants in this 834-bp segment segregate as single haplotype blocks. We propose that these unusually large haplotype blocks are due to positive selection on polymorphisms within the white gene, including a replacement polymorphism, Arg{yields}Leu, within this segment. 45 refs., 4 figs., 1 tab.

  11. COBE ground segment attitude determination

    NASA Technical Reports Server (NTRS)

    Kumar, V. K.; Freedman, I.; Wright, E. L.; Patt, F. S.

    1991-01-01

    The Cosmic Background Explorer (COBE) spacecraft was launched in November 1989 by NASA to survey the sky for primordial radiation left from the Big Bang explosion. The success of the mission requires an accurate determination of the spacecraft attitude. While the accuracy of the attitude obtained from the attitude sensors is adequate for two of the experiments, the higher accuracy required by the Diffuse Infrared Background Experiment (DIRBE) is obtained by using the DIRBE instrument as a special type of star sensor. Presented here is an overview of the attitude processing algorithms used at the Cosmology Data Analysis Center (CDAC) and the results obtained from the flight data.

  12. Patterned Contractile Forces Promote Epidermal Spreading and Regulate Segment Positioning during Drosophila Head Involution.

    PubMed

    Czerniak, Natalia Dorota; Dierkes, Kai; D'Angelo, Arturo; Colombelli, Julien; Solon, Jérôme

    2016-07-25

    Epithelial spreading is a fundamental mode of tissue rearrangement occurring during animal development and wound closure. It has been associated either with the collective migration of cells [1, 2] or with actomyosin-generated forces acting at the leading edge (LE) and pulling the epithelial tissue [3, 4]. During the process of Drosophila head involution (HI), the epidermis spreads anteriorly to envelope the head tissues and fully cover the embryo [5]. This results in epidermal segments of equal width that will give rise to the different organs of the fly [6]. Here we perform a quantitative analysis of tissue spreading during HI. Combining high-resolution live microscopy with laser microsurgery and genetic perturbations, we show that epidermal movement is in part, but not solely, driven by a contractile actomyosin cable at the LE. Additional driving forces are generated within each segment by a gradient of actomyosin-based circumferential tension. Interfering with Hedgehog (Hh) signaling can modulate this gradient, thus suggesting the involvement of polarity genes in the regulation of HI. In particular, we show that disruption of these contractile forces alters segment widths and leads to a mispositioning of segments. Within the framework of a physical description, we confirm that given the geometry of the embryo, a patterned profile of active circumferential tensions can indeed generate propelling forces and control final segment position. Our study thus unravels a mechanism by which patterned tensile forces can regulate spreading and positioning of epithelial tissues. PMID:27397891

  13. Automatic Segmentation of Drosophila Neural Compartments Using GAL4 Expression Data Reveals Novel Visual Pathways.

    PubMed

    Panser, Karin; Tirian, Laszlo; Schulze, Florian; Villalba, Santiago; Jefferis, Gregory S X E; Bühler, Katja; Straw, Andrew D

    2016-08-01

    Identifying distinct anatomical structures within the brain and developing genetic tools to target them are fundamental steps for understanding brain function. We hypothesize that enhancer expression patterns can be used to automatically identify functional units such as neuropils and fiber tracts. We used two recent, genome-scale Drosophila GAL4 libraries and associated confocal image datasets to segment large brain regions into smaller subvolumes. Our results (available at https://strawlab.org/braincode) support this hypothesis because regions with well-known anatomy, namely the antennal lobes and central complex, were automatically segmented into familiar compartments. The basis for the structural assignment is clustering of voxels based on patterns of enhancer expression. These initial clusters are agglomerated to make hierarchical predictions of structure. We applied the algorithm to central brain regions receiving input from the optic lobes. Based on the automated segmentation and manual validation, we can identify and provide promising driver lines for 11 previously identified and 14 novel types of visual projection neurons and their associated optic glomeruli. The same strategy can be used in other brain regions and likely other species, including vertebrates. PMID:27426516

  14. Drosophila 5-HT2 serotonin receptor: coexpression with fushi-tarazu during segmentation.

    PubMed Central

    Colas, J F; Launay, J M; Kellermann, O; Rosay, P; Maroteaux, L

    1995-01-01

    Serotonin, first described as a neurotransmitter in invertebrates, has been investigated mostly for its functions in the mature central nervous system of higher vertebrates. Serotonin receptor diversity has been described in the mammalian brain and in insects. We report the isolation of a cDNA coding for a Drosophila melanogaster serotonin receptor that displays a sequence, a gene organization, and pharmacological properties typical of the mammalian 5-HT2 serotonin receptor subtype. Its mRNA can be detected in the adult fly; moreover, a high level of expression occurs at 3 hr of Drosophila embryogenesis. This early embryonic expression is surprisingly organized in a seven-stripe pattern that appears at the cellular blastoderm stage. In addition, this pattern is in phase with that of the even-parasegment-expressed pair-rule gene fushi-tarazu and is similarly modified by mutations affecting segmentation genes. Simultaneously with this pair-rule expression, the complete machinery of serotonin synthesis is present and leads to a peak of ligand concomitant with a peak of 5-HT2-specific receptor sites in blastoderm embryos. Images Fig. 2 Fig. 3 Fig. 4 PMID:7777527

  15. Ras1-Mediated Modulation of Drosophila Homeotic Function in Cell and Segment Identity

    PubMed Central

    Boube, M.; Benassayag, C.; Seroude, L.; Cribbs, D. L.

    1997-01-01

    Mutations of the Drosophila homeotic proboscipedia gene (pb; the Hox-A2/B2 homologue) provoke dose-sensitive defects. These were used to search for dose-sensitive dominant modifiers of pb function. Two identified interacting genes were the proto-oncogene Ras1 and its functional antagonist Gap1, prominent intermediaries in known signal transduction pathways. Ras1(+) is a positive modifier of pb activity both in normal and ectopic cell contexts, while the Ras1-antagonist Gap1 has an opposite effect. A general role for Ras1 in homeotic function is likely, since Ras1(+) activity also modulates functions of the homeotic loci Sex combs reduced and Ultrabithorax. Our data suggest that the modulation occurs by a mechanism independent of transcriptional control of the homeotic loci themselves, or of the Ras1/Gap1 genes. Taken together our data support a role for Ras1-mediated cell signaling in the homeotic control of segmental differentiation. PMID:9178011

  16. Sequence Analysis of the Segmental Duplication Responsible for Paris Sex-Ratio Drive in Drosophila simulans

    PubMed Central

    Fouvry, Lucie; Ogereau, David; Berger, Anne; Gavory, Frederick; Montchamp-Moreau, Catherine

    2011-01-01

    Sex-ratio distorters are X-linked selfish genetic elements that facilitate their own transmission by subverting Mendelian segregation at the expense of the Y chromosome. Naturally occurring cases of sex-linked distorters have been reported in a variety of organisms, including several species of Drosophila; they trigger genetic conflict over the sex ratio, which is an important evolutionary force. However, with a few exceptions, the causal loci are unknown. Here, we molecularly characterize the segmental duplication involved in the Paris sex-ratio system that is still evolving in natural populations of Drosophila simulans. This 37.5 kb tandem duplication spans six genes, from the second intron of the Trf2 gene (TATA box binding protein-related factor 2) to the first intron of the org-1 gene (optomotor-blind-related-gene-1). Sequence analysis showed that the duplication arose through the production of an exact copy on the template chromosome itself. We estimated this event to be less than 500 years old. We also detected specific signatures of the duplication mechanism; these support the Duplication-Dependent Strand Annealing model. The region at the junction between the two duplicated segments contains several copies of an active transposable element, Hosim1, alternating with 687 bp repeats that are noncoding but transcribed. The almost-complete sequence identity between copies made it impossible to complete the sequencing and assembly of this region. These results form the basis for the functional dissection of Paris sex-ratio drive and will be valuable for future studies designed to better understand the dynamics and the evolutionary significance of sex chromosome drive. PMID:22384350

  17. Drosophila homologs of transcriptional mediator complex subunits are required for adult cell and segment identity specification

    PubMed Central

    Boube, Muriel; Faucher, Christian; Joulia, Laurent; Cribbs, David L.; Bourbon, Henri-Marc

    2000-01-01

    The origins of specificity in gene expression are a central concern in understanding developmental control. Mediator protein complexes regulate transcriptional initiation, acting as modular adaptors linking specific transcription factors to core RNA polymerase II. Here, we identified the Drosophila homologs of 23 human mediator genes and mutations of two, dTRAP240 and of dTRAP80 (the putative fly homolog of yeast SRB4). Clonal analysis indicates a general role for dTRAP80 necessary for cell viability. The dTRAP240 gene is also essential, but cells lacking its function are viable and proliferate normally. Clones reveal localized developmental activities including a sex comb cell identity function. This contrasts with the ubiquitous nuclear accumulation of dTRAP240 protein in imaginal discs. Synergistic genetic interactions support shared developmental cell and segment identity functions of dTRAP240 and dTRAP80, potentially within a common complex. Further, they identify the homeotic Sex combs reduced product, required for the same cell/tissue identities, as a functional partner of these mediator proteins. PMID:11090137

  18. Whole-Embryo Modeling of Early Segmentation in Drosophila Identifies Robust and Fragile Expression Domains

    PubMed Central

    Bieler, Jonathan; Pozzorini, Christian; Naef, Felix

    2011-01-01

    Segmentation of the Drosophila melanogaster embryo results from the dynamic establishment of spatial mRNA and protein patterns. Here, we exploit recent temporal mRNA and protein expression measurements on the full surface of the blastoderm to calibrate a dynamical model of the gap gene network on the entire embryo cortex. We model the early mRNA and protein dynamics of the gap genes hunchback, Kruppel, giant, and knirps, taking as regulatory inputs the maternal Bicoid and Caudal gradients, plus the zygotic Tailless and Huckebein proteins. The model captures the expression patterns faithfully, and its predictions are assessed from gap gene mutants. The inferred network shows an architecture based on reciprocal repression between gap genes that can stably pattern the embryo on a realistic geometry but requires complex regulations such as those involving the Hunchback monomer and dimers. Sensitivity analysis identifies the posterior domain of giant as among the most fragile features of an otherwise robust network, and hints at redundant regulations by Bicoid and Hunchback, possibly reflecting recent evolutionary changes in the gap-gene network in insects. PMID:21767480

  19. Circadian Transcription Contributes to Core Period Determination in Drosophila

    PubMed Central

    Kadener, Sebastian; Menet, Jerome S; Schoer, Rebecca; Rosbash, Michael

    2008-01-01

    The Clock–Cycle (CLK–CYC) heterodimer constitutes a key circadian transcription complex in Drosophila. CYC has a DNA-binding domain but lacks an activation domain. Previous experiments also indicate that most of the transcriptional activity of CLK–CYC derives from the glutamine-rich region of its partner CLK. To address the role of transcription in core circadian timekeeping, we have analyzed the effects of a CYC–viral protein 16 (VP16) fusion protein in the Drosophila system. The addition of this potent and well-studied viral transcriptional activator (VP16) to CYC imparts to the CLK–CYC-VP16 complex strongly enhanced transcriptional activity relative to that of CLK–CYC. This increase is manifested in flies expressing CYC-VP16 as well as in S2 cells. These flies also have increased levels of CLK–CYC direct target gene mRNAs as well as a short period, implicating circadian transcription in period determination. A more detailed examination of reporter gene expression in CYC-VP16–expressing flies suggests that the short period is due at least in part to a more rapid transcriptional phase. Importantly, the behavioral effects require a period (per) promoter and are therefore unlikely to be merely a consequence of generally higher PER levels. This indicates that the CLK–CYC-VP16 behavioral effects are a consequence of increased per transcription. All of this also suggests that the timing of transcriptional activation and not the activation itself is the key event responsible for the behavioral effects observed in CYC-VP16-expressing flies. The results taken together indicate that circadian transcription contributes to core circadian function in Drosophila. PMID:18494558

  20. Segment-specific Ca(2+) transport by isolated Malpighian tubules of Drosophila melanogaster: A comparison of larval and adult stages.

    PubMed

    Browne, Austin; O'Donnell, Michael J

    2016-04-01

    Haemolymph calcium homeostasis in insects is achieved through the regulation of calcium excretion by Malpighian tubules in two ways: (1) sequestration of calcium within biomineralized granules and (2) secretion of calcium in soluble form within the primary urine. Using the scanning ion-selective electrode technique (SIET), basolateral Ca(2+) transport was measured at the distal, transitional, main and proximal tubular segments of anterior tubules isolated from both 3rd instar larvae and adults of the fruit fly Drosophila melanogaster. Basolateral Ca(2+) transport exceeded transepithelial secretion by 800-fold and 11-fold in anterior tubules of larvae and adults, respectively. The magnitude of Ca(2+) fluxes across the distal tubule of larvae and adults were larger than fluxes across the downstream segments by 10 and 40 times, respectively, indicating a dominant role for the distal segment in whole animal Ca(2+) regulation. Basolateral Ca(2+) transport across distal tubules of Drosophila varied throughout the life cycle; Ca(2+) was released by distal tubules of larvae, taken up by distal tubules of young adults and was released once again by tubules of adults ⩾ 168 h post-eclosion. In adults and larvae, SIET measurements revealed sites of both Ca(2+) uptake and Ca(2+) release across the basolateral surface of the distal segment of the same tubule, indicating that Ca(2+) transport is bidirectional. Ca(2+) uptake across the distal segment of tubules of young adults and Ca(2+) release across the distal segment of tubules of older adults was also suggestive of reversible Ca(2+) storage. Our results suggest that the distal tubules of D. melanogaster are dynamic calcium stores which allow efficient haemolymph calcium regulation through active Ca(2+) sequestration during periods of high dietary calcium intake and passive Ca(2+) release during periods of calcium deficiency. PMID:26802560

  1. Maternal AP determinants in the Drosophila oocyte and embryo.

    PubMed

    Ma, Jun; He, Feng; Xie, Gengqiang; Deng, Wu-Min

    2016-09-01

    An animal embryo cannot initiate its journey of forming a new life on its own. It must rely on maternally provided resources and inputs to kick-start its developmental process. In Drosophila, the initial polarities of the embryo along both the anterior-posterior (AP) and dorsal-ventral (DV) axes are also specified by maternal determinants. Over the past several decades, genetic and molecular studies have identified and characterized such determinants, as well as the zygotic genetic regulatory networks that control patterning in the early embryo. Extensive studies of oogenesis have also led to a detailed knowledge of the cellular and molecular interactions that control the formation of a mature egg. Despite these efforts, oogenesis and embryogenesis have been studied largely as separate problems, except for qualitative aspects with regard to maternal regulation of the asymmetric localization of maternal determinants. Can oogenesis and embryogenesis be viewed from a unified perspective at a quantitative level, and can that improve our understanding of how robust embryonic patterning is achieved? Here, we discuss the basic knowledge of the regulatory mechanisms controlling oogenesis and embryonic patterning along the AP axis. We explore properties of the maternal Bicoid gradient in relation to embryo size in search for a unified framework for robust AP patterning. WIREs Dev Biol 2016, 5:562-581. doi: 10.1002/wdev.235 For further resources related to this article, please visit the WIREs website. PMID:27253156

  2. Cell-Autonomous and Non-cell-autonomous Function of Hox Genes Specify Segmental Neuroblast Identity in the Gnathal Region of the Embryonic CNS in Drosophila

    PubMed Central

    Becker, Henrike; Renner, Simone; Technau, Gerhard M.; Berger, Christian

    2016-01-01

    During central nervous system (CNS) development neural stem cells (Neuroblasts, NBs) have to acquire an identity appropriate to their location. In thoracic and abdominal segments of Drosophila, the expression pattern of Bithorax-Complex Hox genes is known to specify the segmental identity of NBs prior to their delamination from the neuroectoderm. Compared to the thoracic, ground state segmental units in the head region are derived to different degrees, and the precise mechanism of segmental specification of NBs in this region is still unclear. We identified and characterized a set of serially homologous NB-lineages in the gnathal segments and used one of them (NB6-4 lineage) as a model to investigate the mechanism conferring segment-specific identities to gnathal NBs. We show that NB6-4 is primarily determined by the cell-autonomous function of the Hox gene Deformed (Dfd). Interestingly, however, it also requires a non-cell-autonomous function of labial and Antennapedia that are expressed in adjacent anterior or posterior compartments. We identify the secreted molecule Amalgam (Ama) as a downstream target of the Antennapedia-Complex Hox genes labial, Dfd, Sex combs reduced and Antennapedia. In conjunction with its receptor Neurotactin (Nrt) and the effector kinase Abelson tyrosine kinase (Abl), Ama is necessary in parallel to the cell-autonomous Dfd pathway for the correct specification of the maxillary identity of NB6-4. Both pathways repress CyclinE (CycE) and loss of function of either of these pathways leads to a partial transformation (40%), whereas simultaneous mutation of both pathways leads to a complete transformation (100%) of NB6-4 segmental identity. Finally, we provide genetic evidences, that the Ama-Nrt-Abl-pathway regulates CycE expression by altering the function of the Hippo effector Yorkie in embryonic NBs. The disclosure of a non-cell-autonomous influence of Hox genes on neural stem cells provides new insight into the process of segmental

  3. Cell-Autonomous and Non-cell-autonomous Function of Hox Genes Specify Segmental Neuroblast Identity in the Gnathal Region of the Embryonic CNS in Drosophila.

    PubMed

    Becker, Henrike; Renner, Simone; Technau, Gerhard M; Berger, Christian

    2016-03-01

    During central nervous system (CNS) development neural stem cells (Neuroblasts, NBs) have to acquire an identity appropriate to their location. In thoracic and abdominal segments of Drosophila, the expression pattern of Bithorax-Complex Hox genes is known to specify the segmental identity of NBs prior to their delamination from the neuroectoderm. Compared to the thoracic, ground state segmental units in the head region are derived to different degrees, and the precise mechanism of segmental specification of NBs in this region is still unclear. We identified and characterized a set of serially homologous NB-lineages in the gnathal segments and used one of them (NB6-4 lineage) as a model to investigate the mechanism conferring segment-specific identities to gnathal NBs. We show that NB6-4 is primarily determined by the cell-autonomous function of the Hox gene Deformed (Dfd). Interestingly, however, it also requires a non-cell-autonomous function of labial and Antennapedia that are expressed in adjacent anterior or posterior compartments. We identify the secreted molecule Amalgam (Ama) as a downstream target of the Antennapedia-Complex Hox genes labial, Dfd, Sex combs reduced and Antennapedia. In conjunction with its receptor Neurotactin (Nrt) and the effector kinase Abelson tyrosine kinase (Abl), Ama is necessary in parallel to the cell-autonomous Dfd pathway for the correct specification of the maxillary identity of NB6-4. Both pathways repress CyclinE (CycE) and loss of function of either of these pathways leads to a partial transformation (40%), whereas simultaneous mutation of both pathways leads to a complete transformation (100%) of NB6-4 segmental identity. Finally, we provide genetic evidences, that the Ama-Nrt-Abl-pathway regulates CycE expression by altering the function of the Hippo effector Yorkie in embryonic NBs. The disclosure of a non-cell-autonomous influence of Hox genes on neural stem cells provides new insight into the process of segmental

  4. Molecular cloning and characterization of human WINS1 and mouse Wins2, homologous to Drosophila segment polarity gene Lines (Lin).

    PubMed

    Katoh, Masaru

    2002-08-01

    WNT signaling molecules play key roles in carcinogenesis and embryogenesis. Drosophila segment polarity gene Lines (Lin) is essential for Wnt/Wingless-dependent patterning in dorsal epidermis and also for hindgut development. With Wnt signaling, Lin accumulates in the nucleus to modulate transcription of Wnt target genes through association with beta-catenin/Armadillo and TCF/Pangolin. Here, human WINS1 and mouse Wins2, encoding proteins with Drosophila Lin homologous domain, were isolated using bioinformatics and cDNA-PCR. Human WINS1 encoded 757-amino-acid protein, and mouse Wins2 encoded 498-amino-acid protein. Human WINS1 and mouse Wins2 showed 60.0% total-amino-acid identity. Lin homologous domain of WINS1 and Wins2 showed 29.4% and 27.2% amino-acid identity with that of Drosphila Lin, respectively. In the human chromosome 15q26 region, WINS1 gene was clustered with ASB7 gene encoding ankyrin repeat and SOCS box-containing protein 7. Human WINS1 mRNA of 2.8-kb in size was expressed in adult testis, prostate, spleen, thymus, skeletal muscle, fetal kidney and brain. This is the first report on molecular cloning and initial characterization of human WINS1 and mouse Wins2 PMID:12119551

  5. Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

    PubMed Central

    Woods, Jared K.; Kowalski, Suzanne; Rogina, Blanka

    2014-01-01

    Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases. PMID:24747955

  6. Determination of the spontaneous locomotor activity in Drosophila melanogaster.

    PubMed

    Woods, Jared K; Kowalski, Suzanne; Rogina, Blanka

    2014-01-01

    Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases. PMID:24747955

  7. Quantitative dynamics and increased variability of segmentation gene expression in the Drosophila Krüppel and knirps mutants.

    PubMed

    Surkova, Svetlana; Golubkova, Elena; Manu; Panok, Lena; Mamon, Lyudmila; Reinitz, John; Samsonova, Maria

    2013-04-01

    Here we characterize the response of the Drosophila segmentation system to mutations in two gap genes, Kr and kni, in the form of single or double homozygotes and single heterozygotes. Segmentation gene expression in these genotypes was quantitatively monitored with cellular resolution in space and 6.5 to 13min resolution in time. As is the case with wild type, we found that gene expression domains in the posterior portion of the embryo shift to the anterior over time. In certain cases, such as the gt posterior domain in Kr mutants, the shifts are significantly larger than is seen in wild type embryos. We also investigated the effects of Kr and kni on the variability of gene expression. Mutations often produce variable phenotypes, and it is well known that the cuticular phenotype of Kr mutants is variable. We sought to understand the molecular basis of this effect. We find that throughout cycle 14A the relative levels of eve and ftz expression in stripes 2 and 3 are variable among individual embryos. Moreover, in Kr and kni mutants, unlike wild type, the variability in positioning of the posterior Hb domain and eve stripe 7 is not decreased or filtered with time. The posterior Gt domain in Kr mutants is highly variable at early times, but this variability decreases when this domain shifts in the anterior direction to the position of the neighboring Kni domain. In contrast to these findings, positional variability throughout the embryo does not decrease over time in double Kr;kni mutants. In heterozygotes the early expression patterns of segmentation genes resemble patterns seen in homozygous mutants but by the onset of gastrulation they become similar to the wild type patterns. Finally, we note that gene expression levels are reduced in Kr and kni mutant embryos and have a tendency to decrease over time. This is a surprising result in view of the role that mutual repression is thought to play in the gap gene system. PMID:23333947

  8. The muscle pattern of the Drosophila abdomen depends on a subdivision of the anterior compartment of each segment

    PubMed Central

    Krzemień, Joanna; Fabre, Caroline C. G.; Casal, José; Lawrence, Peter A.

    2012-01-01

    In the past, segments were defined by landmarks such as muscle attachments, notably by Snodgrass, the king of insect anatomists. Here, we show how an objective definition of a segment, based on developmental compartments, can help explain the dorsal abdomen of adult Drosophila. The anterior (A) compartment of each segment is subdivided into two domains of cells, each responding differently to Hedgehog. The anterior of these domains is non-neurogenic and clones lacking Notch develop normally; this domain can express stripe and form muscle attachments. The posterior domain is neurogenic and clones lacking Notch do not form cuticle; this domain is unable to express stripe or form muscle attachments. The posterior (P) compartment does not form muscle attachments. Our in vivo films indicate that early in the pupa the anterior domain of the A compartment expresses stripe in a narrowing zone that attracts the extending myotubes and resolves into the attachment sites for the dorsal abdominal muscles. We map the tendon cells precisely and show that all are confined to the anterior domain of A. It follows that the dorsal abdominal muscles are intersegmental, spanning from one anterior domain to the next. This view is tested and supported by clones that change cell identity or express stripe ectopically. It seems that growing myotubes originate in posterior A and extend forwards and backwards until they encounter and attach to anterior A cells. The dorsal adult muscles are polarised in the anteroposterior axis: we disprove the hypothesis that muscle orientation depends on genes that define planar cell polarity in the epidermis. PMID:22147953

  9. How to make stripes: deciphering the transition from non-periodic to periodic patterns in Drosophila segmentation.

    PubMed

    Schroeder, Mark D; Greer, Christina; Gaul, Ulrike

    2011-07-01

    The generation of metameric body plans is a key process in development. In Drosophila segmentation, periodicity is established rapidly through the complex transcriptional regulation of the pair-rule genes. The 'primary' pair-rule genes generate their 7-stripe expression through stripe-specific cis-regulatory elements controlled by the preceding non-periodic maternal and gap gene patterns, whereas 'secondary' pair-rule genes are thought to rely on 7-stripe elements that read off the already periodic primary pair-rule patterns. Using a combination of computational and experimental approaches, we have conducted a comprehensive systems-level examination of the regulatory architecture underlying pair-rule stripe formation. We find that runt (run), fushi tarazu (ftz) and odd skipped (odd) establish most of their pattern through stripe-specific elements, arguing for a reclassification of ftz and odd as primary pair-rule genes. In the case of run, we observe long-range cis-regulation across multiple intervening genes. The 7-stripe elements of run, ftz and odd are active concurrently with the stripe-specific elements, indicating that maternal/gap-mediated control and pair-rule gene cross-regulation are closely integrated. Stripe-specific elements fall into three distinct classes based on their principal repressive gap factor input; stripe positions along the gap gradients correlate with the strength of predicted input. The prevalence of cis-elements that generate two stripes and their genomic organization suggest that single-stripe elements arose by splitting and subfunctionalization of ancestral dual-stripe elements. Overall, our study provides a greatly improved understanding of how periodic patterns are established in the Drosophila embryo. PMID:21693522

  10. How to make stripes: deciphering the transition from non-periodic to periodic patterns in Drosophila segmentation

    PubMed Central

    Schroeder, Mark D.; Greer, Christina; Gaul, Ulrike

    2011-01-01

    The generation of metameric body plans is a key process in development. In Drosophila segmentation, periodicity is established rapidly through the complex transcriptional regulation of the pair-rule genes. The ‘primary’ pair-rule genes generate their 7-stripe expression through stripe-specific cis-regulatory elements controlled by the preceding non-periodic maternal and gap gene patterns, whereas ‘secondary’ pair-rule genes are thought to rely on 7-stripe elements that read off the already periodic primary pair-rule patterns. Using a combination of computational and experimental approaches, we have conducted a comprehensive systems-level examination of the regulatory architecture underlying pair-rule stripe formation. We find that runt (run), fushi tarazu (ftz) and odd skipped (odd) establish most of their pattern through stripe-specific elements, arguing for a reclassification of ftz and odd as primary pair-rule genes. In the case of run, we observe long-range cis-regulation across multiple intervening genes. The 7-stripe elements of run, ftz and odd are active concurrently with the stripe-specific elements, indicating that maternal/gap-mediated control and pair-rule gene cross-regulation are closely integrated. Stripe-specific elements fall into three distinct classes based on their principal repressive gap factor input; stripe positions along the gap gradients correlate with the strength of predicted input. The prevalence of cis-elements that generate two stripes and their genomic organization suggest that single-stripe elements arose by splitting and subfunctionalization of ancestral dual-stripe elements. Overall, our study provides a greatly improved understanding of how periodic patterns are established in the Drosophila embryo. PMID:21693522

  11. Cryo-EM Structure Determination Using Segmented Helical Image Reconstruction.

    PubMed

    Fromm, S A; Sachse, C

    2016-01-01

    Treating helices as single-particle-like segments followed by helical image reconstruction has become the method of choice for high-resolution structure determination of well-ordered helical viruses as well as flexible filaments. In this review, we will illustrate how the combination of latest hardware developments with optimized image processing routines have led to a series of near-atomic resolution structures of helical assemblies. Originally, the treatment of helices as a sequence of segments followed by Fourier-Bessel reconstruction revealed the potential to determine near-atomic resolution structures from helical specimens. In the meantime, real-space image processing of helices in a stack of single particles was developed and enabled the structure determination of specimens that resisted classical Fourier helical reconstruction and also facilitated high-resolution structure determination. Despite the progress in real-space analysis, the combination of Fourier and real-space processing is still commonly used to better estimate the symmetry parameters as the imposition of the correct helical symmetry is essential for high-resolution structure determination. Recent hardware advancement by the introduction of direct electron detectors has significantly enhanced the image quality and together with improved image processing procedures has made segmented helical reconstruction a very productive cryo-EM structure determination method. PMID:27572732

  12. Predicting Ancestral Segmentation Phenotypes from Drosophila to Anopheles Using In Silico Evolution.

    PubMed

    Rothschild, Jeremy B; Tsimiklis, Panagiotis; Siggia, Eric D; François, Paul

    2016-05-01

    Molecular evolution is an established technique for inferring gene homology but regulatory DNA turns over so rapidly that inference of ancestral networks is often impossible. In silico evolution is used to compute the most parsimonious path in regulatory space for anterior-posterior patterning linking two Dipterian species. The expression pattern of gap genes has evolved between Drosophila (fly) and Anopheles (mosquito), yet one of their targets, eve, has remained invariant. Our model predicts that stripe 5 in fly disappears and a new posterior stripe is created in mosquito, thus eve stripe modules 3+7 and 4+6 in fly are homologous to 3+6 and 4+5 in mosquito. We can place Clogmia on this evolutionary pathway and it shares the mosquito homologies. To account for the evolution of the other pair-rule genes in the posterior we have to assume that the ancestral Dipterian utilized a dynamic method to phase those genes in relation to eve. PMID:27227405

  13. Predicting Ancestral Segmentation Phenotypes from Drosophila to Anopheles Using In Silico Evolution

    PubMed Central

    Rothschild, Jeremy B.; Tsimiklis, Panagiotis; Siggia, Eric D.; François, Paul

    2016-01-01

    Molecular evolution is an established technique for inferring gene homology but regulatory DNA turns over so rapidly that inference of ancestral networks is often impossible. In silico evolution is used to compute the most parsimonious path in regulatory space for anterior-posterior patterning linking two Dipterian species. The expression pattern of gap genes has evolved between Drosophila (fly) and Anopheles (mosquito), yet one of their targets, eve, has remained invariant. Our model predicts that stripe 5 in fly disappears and a new posterior stripe is created in mosquito, thus eve stripe modules 3+7 and 4+6 in fly are homologous to 3+6 and 4+5 in mosquito. We can place Clogmia on this evolutionary pathway and it shares the mosquito homologies. To account for the evolution of the other pair-rule genes in the posterior we have to assume that the ancestral Dipterian utilized a dynamic method to phase those genes in relation to eve. PMID:27227405

  14. Phf7 controls male sex determination in the Drosophila germline

    PubMed Central

    Yang, Shu Yuan; Baxter, Ellen M.; Van Doren, Mark

    2013-01-01

    Summary Establishment of germline sexual identity is critical for production of male and female germline stem cells, and sperm vs. eggs. Here we identify PHD Finger Protein 7 (PHF7) as an important factor for male germline sexual identity in Drosophila. PHF7 exhibits male-specific expression in early germ cells, germline stem cells and spermatogonia. It is required for germline stem cell maintenance and gametogenesis in males, whereas ectopic expression in female germ cells ablates the germline. Strikingly, expression of PHF7 promotes spermatogenesis in XX germ cells when they are present in a male soma. PHF7 homologs are also specifically expressed in the mammalian testis, and human PHF7 rescues Drosophila Phf7 mutants. PHF7 associates with chromatin and both the human and fly proteins bind histone H3 N-terminal tails with a preference for dimethyl lysine 4 (H3K4me2). We propose that PHF7 acts as a conserved epigenetic “reader” that activates the male germline sexual program. PMID:22595675

  15. Systematic determination of patterns of gene expression during Drosophila embryogenesis

    PubMed Central

    Tomancak, Pavel; Beaton, Amy; Weiszmann, Richard; Kwan, Elaine; Shu, ShengQiang; Lewis, Suzanna E; Richards, Stephen; Ashburner, Michael; Hartenstein, Volker; Celniker, Susan E; Rubin, Gerald M

    2002-01-01

    Background Cell-fate specification and tissue differentiation during development are largely achieved by the regulation of gene transcription. Results As a first step to creating a comprehensive atlas of gene-expression patterns during Drosophila embryogenesis, we examined 2,179 genes by in situ hybridization to fixed Drosophila embryos. Of the genes assayed, 63.7% displayed dynamic expression patterns that were documented with 25,690 digital photomicrographs of individual embryos. The photomicrographs were annotated using controlled vocabularies for anatomical structures that are organized into a developmental hierarchy. We also generated a detailed time course of gene expression during embryogenesis using microarrays to provide an independent corroboration of the in situ hybridization results. All image, annotation and microarray data are stored in publicly available database. We found that the RNA transcripts of about 1% of genes show clear subcellular localization. Nearly all the annotated expression patterns are distinct. We present an approach for organizing the data by hierarchical clustering of annotation terms that allows us to group tissues that express similar sets of genes as well as genes displaying similar expression patterns. Conclusions Analyzing gene-expression patterns by in situ hybridization to whole-mount embryos provides an extremely rich dataset that can be used to identify genes involved in developmental processes that have been missed by traditional genetic analysis. Systematic analysis of rigorously annotated patterns of gene expression will complement and extend the types of analyses carried out using expression microarrays. PMID:12537577

  16. Multiple Genome Segments Determine Virulence of Bluetongue Virus Serotype 8

    PubMed Central

    Janowicz, Anna; Caporale, Marco; Shaw, Andrew; Gulletta, Salvatore; Di Gialleonardo, Luigina; Ratinier, Maxime

    2015-01-01

    ABSTRACT Bluetongue virus (BTV) causes bluetongue, a major hemorrhagic disease of ruminants. In order to investigate the molecular determinants of BTV virulence, we used a BTV8 strain minimally passaged in tissue culture (termed BTV8L in this study) and a derivative strain passaged extensively in tissue culture (BTV8H) in in vitro and in vivo studies. BTV8L was pathogenic in both IFNAR−/− mice and in sheep, while BTV8H was attenuated in both species. To identify genetic changes which led to BTV8H attenuation, we generated 34 reassortants between BTV8L and BTV8H. We found that partial attenuation of BTV8L in IFNAR−/− mice was achieved by simply replacing genomic segment 2 (Seg2, encoding VP2) or Seg10 (encoding NS3) with the BTV8H homologous segments. Fully attenuated viruses required at least two genome segments from BTV8H, including Seg2 with either Seg1 (encoding VP1), Seg6 (encoding VP6 and NS4), or Seg10 (encoding NS3). Conversely, full reversion of virulence of BTV8H required at least five genomic segments of BTV8L. We also demonstrated that BTV8H acquired an increased affinity for glycosaminoglycan receptors during passaging in cell culture due to mutations in its VP2 protein. Replication of BTV8H was relatively poor in interferon (IFN)-competent primary ovine endothelial cells compared to replication of BTV8L, and this phenotype was determined by several viral genomic segments, including Seg4 and Seg9. This study demonstrated that multiple viral proteins contribute to BTV8 virulence. VP2 and NS3 are primary determinants of BTV pathogenesis, but VP1, VP5, VP4, VP6, and VP7 also contribute to virulence. IMPORTANCE Bluetongue is one of the major infectious diseases of ruminants, and it is listed as a notifiable disease by the World Organization for Animal Health (OIE). The clinical outcome of BTV infection varies considerably and depends on environmental and host- and virus-specific factors. Over the years, BTV serotypes/strains with various degrees of

  17. Sensory determinants of behavioral dynamics in Drosophila thermotaxis.

    PubMed

    Klein, Mason; Afonso, Bruno; Vonner, Ashley J; Hernandez-Nunez, Luis; Berck, Matthew; Tabone, Christopher J; Kane, Elizabeth A; Pieribone, Vincent A; Nitabach, Michael N; Cardona, Albert; Zlatic, Marta; Sprecher, Simon G; Gershow, Marc; Garrity, Paul A; Samuel, Aravinthan D T

    2015-01-13

    Complex animal behaviors are built from dynamical relationships between sensory inputs, neuronal activity, and motor outputs in patterns with strategic value. Connecting these patterns illuminates how nervous systems compute behavior. Here, we study Drosophila larva navigation up temperature gradients toward preferred temperatures (positive thermotaxis). By tracking the movements of animals responding to fixed spatial temperature gradients or random temperature fluctuations, we calculate the sensitivity and dynamics of the conversion of thermosensory inputs into motor responses. We discover three thermosensory neurons in each dorsal organ ganglion (DOG) that are required for positive thermotaxis. Random optogenetic stimulation of the DOG thermosensory neurons evokes behavioral patterns that mimic the response to temperature variations. In vivo calcium and voltage imaging reveals that the DOG thermosensory neurons exhibit activity patterns with sensitivity and dynamics matched to the behavioral response. Temporal processing of temperature variations carried out by the DOG thermosensory neurons emerges in distinct motor responses during thermotaxis. PMID:25550513

  18. Sensory determinants of behavioral dynamics in Drosophila thermotaxis

    PubMed Central

    Klein, Mason; Afonso, Bruno; Vonner, Ashley J.; Hernandez-Nunez, Luis; Berck, Matthew; Tabone, Christopher J.; Kane, Elizabeth A.; Pieribone, Vincent A.; Nitabach, Michael N.; Cardona, Albert; Zlatic, Marta; Sprecher, Simon G.; Gershow, Marc; Garrity, Paul A.; Samuel, Aravinthan D. T.

    2015-01-01

    Complex animal behaviors are built from dynamical relationships between sensory inputs, neuronal activity, and motor outputs in patterns with strategic value. Connecting these patterns illuminates how nervous systems compute behavior. Here, we study Drosophila larva navigation up temperature gradients toward preferred temperatures (positive thermotaxis). By tracking the movements of animals responding to fixed spatial temperature gradients or random temperature fluctuations, we calculate the sensitivity and dynamics of the conversion of thermosensory inputs into motor responses. We discover three thermosensory neurons in each dorsal organ ganglion (DOG) that are required for positive thermotaxis. Random optogenetic stimulation of the DOG thermosensory neurons evokes behavioral patterns that mimic the response to temperature variations. In vivo calcium and voltage imaging reveals that the DOG thermosensory neurons exhibit activity patterns with sensitivity and dynamics matched to the behavioral response. Temporal processing of temperature variations carried out by the DOG thermosensory neurons emerges in distinct motor responses during thermotaxis. PMID:25550513

  19. Requirements for Hedgehog, a Segmental Polarity Gene, in Patterning Larval and Adult Cuticle of Drosophila

    PubMed Central

    Mohler, J.

    1988-01-01

    Mutations of the hedgehog gene are generally embryonic lethal, resulting in a lawn of denticles on the ventral surface. In strong alleles, no segmentation is obvious and the anteroposterior polarity of ventral denticles is lost. Temperature shift analysis of a temperature-sensitive allele indicates an embryonic activity period for hedgehog between 2.5 and 6 hr of embryonic development (at 25°) and a larval/pupal period from 4 to 7 days of development (at 25°). Mosaic analysis of hedgehog mutations in the adult cuticle indicates a series of defined defects associated with the failure of appropriate hedgehog expression. In particular, defects in the distal portions of the legs and antenna occur in association with homozygous hedgehog clones in the posterior compartment of those structures. Because the defects are associated with homozygous clones, but are not co-extensive, a type of ``domineering'' nonautonomy is proposed for the activity of the hedgehog gene. PMID:3147217

  20. Mid-Embryo Patterning and Precision in Drosophila Segmentation: Krüppel Dual Regulation of hunchback

    PubMed Central

    Holloway, David M.; Spirov, Alexander V.

    2015-01-01

    In early development, genes are expressed in spatial patterns which later define cellular identities and tissue locations. The mechanisms of such pattern formation have been studied extensively in early Drosophila (fruit fly) embryos. The gap gene hunchback (hb) is one of the earliest genes to be expressed in anterior-posterior (AP) body segmentation. As a transcriptional regulator for a number of downstream genes, the spatial precision of hb expression can have significant effects in the development of the body plan. To investigate the factors contributing to hb precision, we used fine spatial and temporal resolution data to develop a quantitative model for the regulation of hb expression in the mid-embryo. In particular, modelling hb pattern refinement in mid nuclear cleavage cycle 14 (NC14) reveals some of the regulatory contributions of simultaneously-expressed gap genes. Matching the model to recent data from wild-type (WT) embryos and mutants of the gap gene Krüppel (Kr) indicates that a mid-embryo Hb concentration peak important in thoracic development (at parasegment 4, PS4) is regulated in a dual manner by Kr, with low Kr concentration activating hb and high Kr concentration repressing hb. The processes of gene expression (transcription, translation, transport) are intrinsically random. We used stochastic simulations to characterize the noise generated in hb expression. We find that Kr regulation can limit the positional variability of the Hb mid-embryo border. This has been recently corroborated in experimental comparisons of WT and Kr- mutant embryos. Further, Kr regulation can decrease uncertainty in mid-embryo hb expression (i.e. contribute to a smooth Hb boundary) and decrease between-copy transcriptional variability within nuclei. Since many tissue boundaries are first established by interactions between neighbouring gene expression domains, these properties of Hb-Kr dynamics to diminish the effects of intrinsic expression noise may represent a

  1. Select interneuron clusters determine female sexual receptivity in Drosophila

    PubMed Central

    Sakurai, Akira; Koganezawa, Masayuki; Yasunaga, Kei-ichiro; Emoto, Kazuo; Yamamoto, Daisuke

    2013-01-01

    Female Drosophila with the spinster mutation repel courting males and rarely mate. Here we show that the non-copulating phenotype can be recapitulated by the elimination of spinster functions from either spin-A or spin-D neuronal clusters, in the otherwise wild-type (spinster heterozygous) female brain. Spin-D corresponds to the olfactory projection neurons with dendrites in the antennal lobe VA1v glomerulus that is fruitless-positive, sexually dimorphic and responsive to fly odour. Spin-A is a novel local neuron cluster in the suboesophageal ganglion, which is known to process contact chemical pheromone information and copulation-related signals. A slight reduction in spinster expression to a level with a minimal effect is sufficient to shut off female sexual receptivity if the dominant-negative mechanistic target of rapamycin is simultaneously expressed, although the latter manipulation alone has only a marginal effect. We propose that spin-mediated mechanistic target of rapamycin signal transduction in these neurons is essential for females to accept the courting male. PMID:23652013

  2. Neutron Resonance Spin Determination Using Multi-Segmented Detector DANCE

    SciTech Connect

    Baramsai, B.; Mitchell, G. E.; Chyzh, A.; Dashdorj, D.; Walker, C.; Agvaanluvsan, U.; Becvar, F.; Krticka, M.; Bredeweg, T. A.; Couture, A.; Haight, R. C.; Jandel, M.; Keksis, A. L.; O'Donnell, J. M.; Rundberg, R. S.; Ullmann, J. L.; Vieira, D. J.; Wouters, J. M.

    2011-06-01

    A sensitive method to determine the spin of neutron resonances is introduced based on the statistical pattern recognition technique. The new method was used to assign the spins of s-wave resonances in {sup 155}Gd. The experimental neutron capture data for these nuclei were measured with the DANCE (Detector for Advanced Neutron Capture Experiment) calorimeter at the Los Alamos Neutron Science Center. The highly segmented calorimeter provided detailed multiplicity distributions of the capture {gamma}-rays. Using this information, the spins of the neutron capture resonances were determined. With these new spin assignments, level spacings are determined separately for s-wave resonances with J{sup {pi}} = 1{sup -} and 2{sup -}.

  3. Altering the sex determination pathway in Drosophila fat body modifies sex-specific stress responses

    PubMed Central

    Neckameyer, Wendi S.

    2014-01-01

    The stress response in Drosophila melanogaster reveals sex differences in behavior, similar to what has been observed in mammals. However, unlike mammals, the sex determination pathway in Drosophila is well established, making this an ideal system to identify factors involved in the modulation of sex-specific responses to stress. In this study, we show that the Drosophila fat body, which has been shown to be important for energy homeostasis and sex determination, is a dynamic tissue that is altered in response to stress in a sex and time-dependent manner. We manipulated the sex determination pathway in the fat body via targeted expression of transformer and transformer-2 and analyzed these animals for changes in their response to stress. In the majority of cases, manipulation of transformer or transformer-2 was able to change the physiological output in response to starvation and oxidative stress to that of the opposite sex. Our data also uncover the possibility of additional downstream targets for transformer and transformer-2 that are separate from the sex determination pathway and can influence behavioral and physiological responses. PMID:24789992

  4. Differential Selection within the Drosophila Retinal Determination Network and Evidence for Functional Divergence between Paralog Pairs

    PubMed Central

    Datta, Rhea R.; Cruickshank, Tami; Kumar, Justin P.

    2011-01-01

    The retinal determination (RD) network in Drosophila comprises fourteen known nuclear proteins that include DNA binding proteins, transcriptional co-activators, kinases and phosphatases. The composition of the network varies considerably throughout the animal kingdom, with the network in several basal insects having fewer members and with vertebrates having potentially significantly higher numbers of retinal determination genes. One important contributing factor for the variation in gene number within the network is gene duplication. For example, ten members of the RD network in Drosophila are derived from duplication events. Here we present an analysis of the coding regions of the five pairs of duplicate genes from within the retinal determination network of several different Drosophila species. We demonstrate that there is differential selection across the coding regions of all RD genes. Additionally, some of the most significant differences in ratios of non-silent to silent site substitutions (dN/dS) between paralog pairs are found within regions that have no ascribed function. Previous structure/function analyses of several duplicate genes have identified areas within one gene that contain novel activities when compared to its paralog. The evolutionary analysis presented here identifies these same areas in the paralogs as being under high levels of relaxed selection. We suggest that sequence divergence between paralogs and selection signatures can be used as a reasonable predictor of functional changes in rapidly evolving motifs. PMID:21210943

  5. Sex-lethal, master and slave: a hierarchy of germ-line sex determination in Drosophila.

    PubMed

    Oliver, B; Kim, Y J; Baker, B S

    1993-11-01

    Female sex determination in the germ line of Drosophila melanogaster is regulated by genes functioning in the soma as well as genes that function within the germ line. Genes known or suspected to be involved in germ-line sex determination in Drosophila melanogaster have been examined to determine if they are required upstream or downstream of Sex-lethal+, a known germ-line sex determination gene. Seven genes required for female-specific splicing of germ-line Sex-lethal+ pre-mRNA are identified. These results together with information about the tissues in which these genes function and whether they control sex determination and viability or just sex determination in the germ line have been used to deduce the genetic hierarchy regulating female germ-line sex determination. This hierarchy includes the somatic sex determination genes transformer+, transformer-2+ and doublesex+ (and by inference Sex-lethal+), which control a somatic signal required for female germ-line sex determination, and the germ-line ovarian tumor genes fused+, ovarian tumor+, ovo+, sans fille+, and Sex-lethal+, which are involved in either the reception or interpretation of this somatic sex determination signal. The fused+, ovarian tumor+, ovo+ and sans fille+ genes function upstream of Sex-lethal+ in the germ line. PMID:8187645

  6. Segments.

    ERIC Educational Resources Information Center

    Zemsky, Robert; Shaman, Susan; Shapiro, Daniel B.

    2001-01-01

    Presents a market taxonomy for higher education, including what it reveals about the structure of the market, the model's technical attributes, and its capacity to explain pricing behavior. Details the identification of the principle seams separating one market segment from another and how student aspirations help to organize the market, making…

  7. Early embryonic determination of the sexual dimorphism in segment number in geophilomorph centipedes

    PubMed Central

    2013-01-01

    Background Most geophilomorph centipedes show intraspecific variability in the number of leg-bearing segments. This intraspecific variability generally has a component that is related to sex, with females having on average more segments than males. Neither the developmental basis nor the adaptive role of this dimorphism is known. Results To determine when this sexual dimorphism in segment number is established, we have followed the development of Strigamia maritima embryos from the onset of segmentation to the first post-embryonic stage where we could determine the sex morphologically. We find that males and females differ in segment number by Stage 6.1, a point during embryogenesis when segment addition pauses while the embryo undergoes large-scale movements. We have confirmed this pattern by establishing a molecular method to determine the sex of single embryos, utilising duplex PCR amplification for Y chromosomal and autosomal sequences. This confirms that male embryos have a modal number of 43 segments visible at Stage 6, while females have 45. In our Strigamia population, adult males have a modal number of 47 leg-bearing segments, and females have 49. This implies that the sexual dimorphism in segment number is determined before the addition of the last leg-bearing segments and the terminal genital segments. Conclusions Sexual dimorphism in segment number is not associated with terminal segment differentiation, but must instead be related to some earlier process during segment patterning. The dimorphism may be associated with a difference in the rate and/or duration of segment addition during the main phase of rapid segment addition that precedes embryonic Stage 6. This suggests that the adaptive role, if any, of the dimorphism is likely to be related to segment number per se, and not to sexual differentiation of the terminal region. PMID:23919293

  8. Estimates of Gene Flow in Drosophila Pseudoobscura Determined from Nucleotide Sequence Analysis of the Alcohol Dehydrogenase Region

    PubMed Central

    Schaeffer, S. W.; Miller, E. L.

    1992-01-01

    The genetic structure of Drosophila pseudoobscura populations was inferred from a nucleotide sequence analysis of a 3.4-kb segment of the alcohol dehydrogenase (Adh) region. A total of 99 isochromosomal strains collected from 13 populations in North and South America were used to determine if any population departed from a neutral model and to estimate levels of gene flow between populations. This study also included the nucleotide sequences from two sibling species, D. persimilis and D. miranda. We estimated the neutral mutation parameter, 4Nμ, in synonymous and noncoding sites for 17 subregions of Adh in each of nine populations with sample sizes greater than three. The nucleotide diversity data in the nine populations was tested for departures from an equilibrium neutral model with two statistical tests. The Tajima and the Hudson, Kreitman, Aguade tests showed that each population fails to reject a neutral model. Tests for genetic differentiation between populations fail to show any population substructure among the North American populations of D. pseudoobscura. The nucleotide diversity data is consistent with direct and indirect measures of gene flow that show extensive dispersal between populations of D. pseudoobscura. PMID:1427038

  9. Molecular Genetics of the Drosophila Melanogaster Ovo Locus, a Gene Required for Sex Determination of Germline Cells

    PubMed Central

    Garfinkel, M. D.; Lohe, A. R.; Mahowald, A. P.

    1992-01-01

    The Drosophila melanogaster ovo gene is required for survival and differentiation of female germline cells, apparently playing a role in germline sex determination. We recovered 60 kb of genomic DNA from its genetic location at 4E1,2 on the X chromosome. A transcription unit coding for an apparently female-specific germline-dependent 5-kb poly(A)(+) RNA size class is located substantially in a 7-kb region, within which three DNA-detectable lesions for mutations that inactivate the ovo function are located at two sites &4 kb apart. The breakpoint of a deficiency that removes the neighboring lethal complementation group shavenbaby (svb) but leaves the ovo function intact maps &5 kb to the molecular left of the leftmost ovo mutant site. A class of mutations that inactivates both the svb function and the ovo function affects genomic DNA between the two ovo sites. Sequences required for the two genetic functions are partly overlapping. In spite of this overlap, P element-mediated gene transfer of a 10-kb genomic DNA segment containing the 5-kb poly(A)(+) RNA transcription unit rescues the female sterility phenotypes of ovo mutations, but not the svb lethality. PMID:1349870

  10. The Sex Determination Gene transformer Regulates Male-Female Differences in Drosophila Body Size

    PubMed Central

    Rideout, Elizabeth J.; Narsaiya, Marcus S.; Grewal, Savraj S.

    2015-01-01

    Almost all animals show sex differences in body size. For example, in Drosophila, females are larger than males. Although Drosophila is widely used as a model to study growth, the mechanisms underlying this male-female difference in size remain unclear. Here, we describe a novel role for the sex determination gene transformer (tra) in promoting female body growth. Normally, Tra is expressed only in females. We find that loss of Tra in female larvae decreases body size, while ectopic Tra expression in males increases body size. Although we find that Tra exerts autonomous effects on cell size, we also discovered that Tra expression in the fat body augments female body size in a non cell-autonomous manner. These effects of Tra do not require its only known targets doublesex and fruitless. Instead, Tra expression in the female fat body promotes growth by stimulating the secretion of insulin-like peptides from insulin producing cells in the brain. Our data suggest a model of sex-specific growth in which body size is regulated by a previously unrecognized branch of the sex determination pathway, and identify Tra as a novel link between sex and the conserved insulin signaling pathway. PMID:26710087

  11. The Sex Determination Gene transformer Regulates Male-Female Differences in Drosophila Body Size.

    PubMed

    Rideout, Elizabeth J; Narsaiya, Marcus S; Grewal, Savraj S

    2015-12-01

    Almost all animals show sex differences in body size. For example, in Drosophila, females are larger than males. Although Drosophila is widely used as a model to study growth, the mechanisms underlying this male-female difference in size remain unclear. Here, we describe a novel role for the sex determination gene transformer (tra) in promoting female body growth. Normally, Tra is expressed only in females. We find that loss of Tra in female larvae decreases body size, while ectopic Tra expression in males increases body size. Although we find that Tra exerts autonomous effects on cell size, we also discovered that Tra expression in the fat body augments female body size in a non cell-autonomous manner. These effects of Tra do not require its only known targets doublesex and fruitless. Instead, Tra expression in the female fat body promotes growth by stimulating the secretion of insulin-like peptides from insulin producing cells in the brain. Our data suggest a model of sex-specific growth in which body size is regulated by a previously unrecognized branch of the sex determination pathway, and identify Tra as a novel link between sex and the conserved insulin signaling pathway. PMID:26710087

  12. Birth order dependent growth cone segregation determines synaptic layer identity in the Drosophila visual system

    PubMed Central

    Kulkarni, Abhishek; Ertekin, Deniz; Lee, Chi-Hon; Hummel, Thomas

    2016-01-01

    The precise recognition of appropriate synaptic partner neurons is a critical step during neural circuit assembly. However, little is known about the developmental context in which recognition specificity is important to establish synaptic contacts. We show that in the Drosophila visual system, sequential segregation of photoreceptor afferents, reflecting their birth order, lead to differential positioning of their growth cones in the early target region. By combining loss- and gain-of-function analyses we demonstrate that relative differences in the expression of the transcription factor Sequoia regulate R cell growth cone segregation. This initial growth cone positioning is consolidated via cell-adhesion molecule Capricious in R8 axons. Further, we show that the initial growth cone positioning determines synaptic layer selection through proximity-based axon-target interactions. Taken together, we demonstrate that birth order dependent pre-patterning of afferent growth cones is an essential pre-requisite for the identification of synaptic partner neurons during visual map formation in Drosophila. DOI: http://dx.doi.org/10.7554/eLife.13715.001 PMID:26987017

  13. Consumption of dietary sugar by gut bacteria determines Drosophila lipid content.

    PubMed

    Huang, Jia-Hsin; Douglas, Angela E

    2015-09-01

    Gut microorganisms are essential for the nutritional health of many animals, but the underlying mechanisms are poorly understood. This study investigated how lipid accumulation by adult Drosophila melanogaster is reduced in flies associated with the bacterium Acetobacter tropicalis which displays oral-faecal cycling between the gut and food. We demonstrate that the lower lipid content of A. tropicalis-colonized flies relative to bacteria-free flies is linked with a parallel bacterial-mediated reduction in food glucose content; and can be accounted for quantitatively by the amount of glucose acquired by the flies, as determined from the feeding rate and assimilation efficiency of bacteria-free and A. tropicalis-colonized flies. We recommend that nutritional studies on Drosophila include empirical quantification of food nutrient content, to account for likely microbial-mediated effects on diet composition. More broadly, this study demonstrates that selective consumption of dietary constituents by microorganisms can alter the nutritional balance of food and, thereby, influence the nutritional status of the animal host. PMID:26382071

  14. Birth order dependent growth cone segregation determines synaptic layer identity in the Drosophila visual system.

    PubMed

    Kulkarni, Abhishek; Ertekin, Deniz; Lee, Chi-Hon; Hummel, Thomas

    2016-01-01

    The precise recognition of appropriate synaptic partner neurons is a critical step during neural circuit assembly. However, little is known about the developmental context in which recognition specificity is important to establish synaptic contacts. We show that in the Drosophila visual system, sequential segregation of photoreceptor afferents, reflecting their birth order, lead to differential positioning of their growth cones in the early target region. By combining loss- and gain-of-function analyses we demonstrate that relative differences in the expression of the transcription factor Sequoia regulate R cell growth cone segregation. This initial growth cone positioning is consolidated via cell-adhesion molecule Capricious in R8 axons. Further, we show that the initial growth cone positioning determines synaptic layer selection through proximity-based axon-target interactions. Taken together, we demonstrate that birth order dependent pre-patterning of afferent growth cones is an essential pre-requisite for the identification of synaptic partner neurons during visual map formation in Drosophila. PMID:26987017

  15. Dpp signaling determines regional stem cell identity in the regenerating adult Drosophila gastrointestinal tract

    PubMed Central

    Li, Hongjie; Qi, Yanyan; Jasper, Heinrich

    2013-01-01

    Summary The gastrointestinal tract is lined by a series of epithelia that share functional requirements, but also have distinct, highly specialized roles. Distinct populations of somatic stem cells (SCs) regenerate these epithelia, yet the mechanisms that maintain regional identities of these SCs are not well understood. Here, we identify a role for the BMP-like Dpp signaling pathway in diversifying regenerative processes in the adult gastrointestinal tract of Drosophila. Dpp secreted from enterocytes at the boundary between the posterior midgut (PM) and the middle midgut (MM) sets up a morphogen gradient that selectively directs copper cell (CC) regeneration from gastric SCs in the MM and thus determines the size of the CC region. In vertebrates, deregulation of BMP signaling has been associated with Barrett’s metaplasia, where the squamous esophageal epithelium is replaced by a columnar epithelium, suggesting that the maintenance of regional SC identities by BMP is conserved. PMID:23810561

  16. Metagenome-Wide Association of Microbial Determinants of Host Phenotype in Drosophila melanogaster

    PubMed Central

    Newell, Peter D.; Douglas, Angela E.

    2014-01-01

    ABSTRACT Animal-associated bacteria (microbiota) affect host behaviors and physiological traits. To identify bacterial genetic determinants of microbiota-responsive host traits, we employed a metagenome-wide association (MGWA) approach in two steps. First, we measured two microbiota-responsive host traits, development time and triglyceride (TAG) content, in Drosophila melanogaster flies monoassociated with each of 41 bacterial strains. The effects of monoassociation on host traits were not confined to particular taxonomic groups. Second, we clustered protein-coding sequences of the bacteria by sequence similarity de novo and statistically associated the magnitude of the host trait with the bacterial gene contents. The animals had been monoassociated with genome-sequenced bacteria, so the metagenome content was unambiguous. This analysis showed significant effects of pyrroloquinoline quinone biosynthesis genes on development time, confirming the results of a published transposon mutagenesis screen, thereby validating the MGWA; it also identified multiple genes predicted to affect host TAG content, including extracellular glucose oxidation pathway components. To test the validity of the statistical associations, we expressed candidate genes in a strain that lacks them. Monoassociation with bacteria that ectopically expressed a predicted oxidoreductase or gluconate dehydrogenase conferred reduced Drosophila TAG contents relative to the TAG contents in empty vector controls. Consistent with the prediction that glucose oxidation pathway gene expression increased bacterial glucose utilization, the glucose content of the host diet was reduced when flies were exposed to these strains. Our findings indicate that microbiota affect host nutritional status through modulation of nutrient acquisition. Together, these findings demonstrate the utility of MGWA for identifying bacterial determinants of host traits and provide mechanistic insight into how gut microbiota modulate the

  17. The hierarchical relation between X-chromosomes and autosomal sex determining genes in Drosophila

    PubMed Central

    Steinmann-Zwicky, Monica; Nöthiger, Rolf

    1985-01-01

    The classical balance concept of sex determination in Drosophila states that the X-chromosome carries dispersed female-determining factors. Besides, a number of autosomal genes are known that, when mutant, transform chromosomal females (XX) into pseudomales (tra), or intersexes (ix, dsx, dsxD). To test whether large duplications of the X-chromosome have a feminizing effect on the sexual phenotype of these mutants, we constructed flies that were mutant for ix, dsx, dsxD or tra and had two X-chromosomes plus either a distal or a proximal half of an X-chromosome. These or even smaller X-chromosomal fragments had a strong feminizing effect when added to triploid intersexes (XX; AAA). In the mutants, however, no shift towards femaleness was apparent. We conclude that enhancing the female determining signal is ineffective in flies that are mutant for an autosomal sex determining gene, and therefore, that these genes are under hierarchical control of the signal given by the X:A ratio. Parallels between sex-determining and homeotic genes are drawn. ImagesFig. 3. PMID:16453598

  18. The origin of chromosomal inversions as a source of segmental duplications in the Sophophora subgenus of Drosophila

    PubMed Central

    Puerma, Eva; Orengo, Dorcas J.; Aguadé, Montserrat

    2016-01-01

    Chromosomal inversions can contribute to the adaptation of organisms to their environment by capturing particular advantageous allelic combinations of a set of genes included in the inverted fragment and also by advantageous functional changes due to the inversion process itself that might affect not only the expression of flanking genes but also their dose and structure. Of the two mechanisms originating inversions —ectopic recombination, and staggered double-strand breaks and subsequent repair— only the latter confers the inversion the potential to have dosage effects and/or to generate advantageous chimeric genes. In Drosophila subobscura, there is ample evidence for the adaptive character of its chromosomal polymorphism, with an important contribution of some warm-climate arrangements such as E1+2+9+12. Here, we have characterized the breakpoints of inversion E12 and established that it originated through the staggered-break mechanism like four of the five inversions of D. subobscura previously studied. This mechanism that also predominates in the D. melanogaster lineage might be prevalent in the Sophophora subgenus and contribute to the adaptive character of the polymorphic and fixed inversions of its species. Finally, we have shown that the D. subobscura inversion breakpoint regions have generally been disrupted by additional structural changes occurred at different time scales. PMID:27470196

  19. Ultra-deep profiling of alternatively spliced Drosophila Dscam isoforms by circularization-assisted multi-segment sequencing

    PubMed Central

    Sun, Wei; You, Xintian; Gogol-Döring, Andreas; He, Haihuai; Kise, Yoshiaki; Sohn, Madlen; Chen, Tao; Klebes, Ansgar; Schmucker, Dietmar; Chen, Wei

    2013-01-01

    The Drosophila melanogaster gene Dscam (Down syndrome cell adhesion molecule) can generate thousands of different ectodomains via mutual exclusive splicing of three large exon clusters. The isoform diversity plays a profound role in both neuronal wiring and pathogen recognition. However, the isoform expression pattern at the global level remained unexplored. Here, we developed a novel method that allows for direct quantification of the alternatively spliced exon combinations from over hundreds of millions of Dscam transcripts in one sequencing run. With unprecedented sequencing depth, we detected a total of 18 496 isoforms, out of 19 008 theoretically possible combinations. Importantly, we demonstrated that alternative splicing between different clusters is independent. Moreover, the isoforms were expressed across a broad dynamic range, with significant bias in cell/tissue and developmental stage-specific patterns. Hitherto underappreciated, such bias can dramatically reduce the ability of neurons to display unique surface receptor codes. Therefore, the seemingly excessive diversity encoded in the Dscam locus might nevertheless be essential for a robust self and non-self discrimination in neurons. PMID:23792425

  20. The origin of chromosomal inversions as a source of segmental duplications in the Sophophora subgenus of Drosophila.

    PubMed

    Puerma, Eva; Orengo, Dorcas J; Aguadé, Montserrat

    2016-01-01

    Chromosomal inversions can contribute to the adaptation of organisms to their environment by capturing particular advantageous allelic combinations of a set of genes included in the inverted fragment and also by advantageous functional changes due to the inversion process itself that might affect not only the expression of flanking genes but also their dose and structure. Of the two mechanisms originating inversions -ectopic recombination, and staggered double-strand breaks and subsequent repair- only the latter confers the inversion the potential to have dosage effects and/or to generate advantageous chimeric genes. In Drosophila subobscura, there is ample evidence for the adaptive character of its chromosomal polymorphism, with an important contribution of some warm-climate arrangements such as E1+2+9+12. Here, we have characterized the breakpoints of inversion E12 and established that it originated through the staggered-break mechanism like four of the five inversions of D. subobscura previously studied. This mechanism that also predominates in the D. melanogaster lineage might be prevalent in the Sophophora subgenus and contribute to the adaptive character of the polymorphic and fixed inversions of its species. Finally, we have shown that the D. subobscura inversion breakpoint regions have generally been disrupted by additional structural changes occurred at different time scales. PMID:27470196

  1. Genetic Evidence That the Ovo Locus Is Involved in Drosophila Germ Line Sex Determination

    PubMed Central

    Oliver, B.; Pauli, D.; Mahowald, A. P.

    1990-01-01

    Zygotically contributed ovo gene product is required for the survival of female germ cells in Drosophila melanogaster. Trans-allelic combinations of weak and dominant ovo mutations (ovo(D)) result in viable germ cells that appear to be partially transformed from female to male sexual identity. The ovo(D2) mutation is partially suppressed by many Sex-lethal alleles that affect the soma, while those that affect only the germ line fail to interact with ovo(D2). One of two loss-of-function ovo alleles is suppressed by a loss-of-function Sex-lethal allele. Because ovo mutations are germ line dependent, it is likely that ovo is suppressed by way of communication between the somatic and germ lines. A loss-of-function allele of ovo is epistatic to germ line dependent mutations in Sex-lethal. The germ line dependent sex determination mutation, sans fille, and ovo(D) mutations show a dominant synergistic interaction resulting in partial transformation of germ line sexual identity. The ovo locus appears to be involved in germ line sex determination and is linked in some manner to sex determination in the soma. PMID:2116356

  2. Parameter estimation and determinability analysis applied to Drosophila gap gene circuits

    PubMed Central

    Ashyraliyev, Maksat; Jaeger, Johannes; Blom, Joke G

    2008-01-01

    Background Mathematical modeling of real-life processes often requires the estimation of unknown parameters. Once the parameters are found by means of optimization, it is important to assess the quality of the parameter estimates, especially if parameter values are used to draw biological conclusions from the model. Results In this paper we describe how the quality of parameter estimates can be analyzed. We apply our methodology to assess parameter determinability for gene circuit models of the gap gene network in early Drosophila embryos. Conclusion Our analysis shows that none of the parameters of the considered model can be determined individually with reasonable accuracy due to correlations between parameters. Therefore, the model cannot be used as a tool to infer quantitative regulatory weights. On the other hand, our results show that it is still possible to draw reliable qualitative conclusions on the regulatory topology of the gene network. Moreover, it improves previous analyses of the same model by allowing us to identify those interactions for which qualitative conclusions are reliable, and those for which they are ambiguous. PMID:18817540

  3. Determination of lung segments in computed tomography images using the Euclidean distance to the pulmonary artery

    SciTech Connect

    Stoecker, Christina; Moltz, Jan H.; Lassen, Bianca; Kuhnigk, Jan-Martin; Krass, Stefan; Welter, Stefan; Peitgen, Heinz-Otto

    2013-09-15

    Purpose: Computed tomography (CT) imaging is the modality of choice for lung cancer diagnostics. With the increasing number of lung interventions on sublobar level in recent years, determining and visualizing pulmonary segments in CT images and, in oncological cases, reliable segment-related information about the location of tumors has become increasingly desirable. Computer-assisted identification of lung segments in CT images is subject of this work.Methods: The authors present a new interactive approach for the segmentation of lung segments that uses the Euclidean distance of each point in the lung to the segmental branches of the pulmonary artery. The aim is to analyze the potential of the method. Detailed manual pulmonary artery segmentations are used to achieve the best possible segment approximation results. A detailed description of the method and its evaluation on 11 CT scans from clinical routine are given.Results: An accuracy of 2–3 mm is measured for the segment boundaries computed by the pulmonary artery-based method. On average, maximum deviations of 8 mm are observed. 135 intersegmental pulmonary veins detected in the 11 test CT scans serve as reference data. Furthermore, a comparison of the presented pulmonary artery-based approach to a similar approach that uses the Euclidean distance to the segmental branches of the bronchial tree is presented. It shows a significantly higher accuracy for the pulmonary artery-based approach in lung regions at least 30 mm distal to the lung hilum.Conclusions: A pulmonary artery-based determination of lung segments in CT images is promising. In the tests, the pulmonary artery-based determination has been shown to be superior to the bronchial tree-based determination. The suitability of the segment approximation method for application in the planning of segment resections in clinical practice has already been verified in experimental cases. However, automation of the method accompanied by an evaluation on a larger

  4. Genetic effects induced by neutrons in Drosophila melanogaster I. Determination of absorbed dose.

    PubMed

    Delfin, A; Paredes, L C; Zambrano, F; Guzmán-Rincón, J; Ureña-Nuñez, F

    2001-12-01

    A method to obtain the absorbed dose in Drosophila melanogaster irradiated in the thermal column facility of the Triga Mark III Reactor has been developed. The method is based on the measurements of neutron activation of gold foils produced by neutron capture to obtain the neutron fluxes. These fluxes, combined with the calculations of kinetic energy released per unit mass, enables one to obtain the absorbed doses in Drosophila melanogaster. PMID:11761104

  5. Retinal determination genes coordinate neuroepithelial specification and neurogenesis modes in the Drosophila optic lobe.

    PubMed

    Apitz, Holger; Salecker, Iris

    2016-07-01

    Differences in neuroepithelial patterning and neurogenesis modes contribute to area-specific diversifications of neural circuits. In the Drosophila visual system, two neuroepithelia, the outer (OPC) and inner (IPC) proliferation centers, generate neuron subtypes for four ganglia in several ways. Whereas neuroepithelial cells in the medial OPC directly convert into neuroblasts, in an IPC subdomain they generate migratory progenitors by epithelial-mesenchymal transition that mature into neuroblasts in a second proliferative zone. The molecular mechanisms that regulate the identity of these neuroepithelia, including their neurogenesis modes, remain poorly understood. Analysis of Polycomblike revealed that loss of Polycomb group-mediated repression of the Hox gene Abdominal-B (Abd-B) caused the transformation of OPC to IPC neuroepithelial identity. This suggests that the neuroepithelial default state is IPC-like, whereas OPC identity is derived. Ectopic Abd-B blocks expression of the highly conserved retinal determination gene network members Eyes absent (Eya), Sine oculis (So) and Homothorax (Hth). These factors are essential for OPC specification and neurogenesis control. Finally, eya and so are also sufficient to confer OPC-like identity, and, in parallel with hth, the OPC-specific neurogenesis mode on the IPC. PMID:27381228

  6. Retinal determination genes coordinate neuroepithelial specification and neurogenesis modes in the Drosophila optic lobe

    PubMed Central

    Apitz, Holger

    2016-01-01

    Differences in neuroepithelial patterning and neurogenesis modes contribute to area-specific diversifications of neural circuits. In the Drosophila visual system, two neuroepithelia, the outer (OPC) and inner (IPC) proliferation centers, generate neuron subtypes for four ganglia in several ways. Whereas neuroepithelial cells in the medial OPC directly convert into neuroblasts, in an IPC subdomain they generate migratory progenitors by epithelial-mesenchymal transition that mature into neuroblasts in a second proliferative zone. The molecular mechanisms that regulate the identity of these neuroepithelia, including their neurogenesis modes, remain poorly understood. Analysis of Polycomblike revealed that loss of Polycomb group-mediated repression of the Hox gene Abdominal-B (Abd-B) caused the transformation of OPC to IPC neuroepithelial identity. This suggests that the neuroepithelial default state is IPC-like, whereas OPC identity is derived. Ectopic Abd-B blocks expression of the highly conserved retinal determination gene network members Eyes absent (Eya), Sine oculis (So) and Homothorax (Hth). These factors are essential for OPC specification and neurogenesis control. Finally, eya and so are also sufficient to confer OPC-like identity, and, in parallel with hth, the OPC-specific neurogenesis mode on the IPC. PMID:27381228

  7. Directed Screen of Francisella novicida Virulence Determinants Using Drosophila melanogaster▿ †

    PubMed Central

    Åhlund, Monika K.; Rydén, Patrik; Sjöstedt, Anders; Stöven, Svenja

    2010-01-01

    Francisella tularensis is a highly virulent, facultative intracellular human pathogen whose virulence mechanisms are not well understood. Occasional outbreaks of tularemia and the potential use of F. tularensis as a bioterrorist agent warrant better knowledge about the pathogenicity of this bacterium. Thus far, genome-wide in vivo screens for virulence factors have been performed in mice, all however restricted by the necessity to apply competition-based, negative-selection assays. We wanted to individually evaluate putative virulence determinants suggested by such assays and performed directed screening of 249 F. novicida transposon insertion mutants by using survival of infected fruit flies as a measure of bacterial virulence. Some 20% of the genes tested were required for normal virulence in flies; most of these had not previously been investigated in detail in vitro or in vivo. We further characterized their involvement in bacterial proliferation and pathogenicity in flies and in mouse macrophages. Hierarchical cluster analysis of mutant phenotypes indicated a functional linkage between clustered genes. One cluster grouped all but four genes of the Francisella pathogenicity island and other loci required for intracellular survival. We also identified genes involved in adaptation to oxidative stress and genes which might induce host energy wasting. Several genes related to type IV pilus formation demonstrated hypervirulent mutant phenotypes. Collectively, the data demonstrate that the bacteria in part use similar virulence mechanisms in mammals as in Drosophila melanogaster but that a considerable proportion of the virulence factors active in mammals are dispensable for pathogenicity in the insect model. PMID:20479082

  8. An Interactive network of long non-coding RNAs facilitates the Drosophila sex determination decision

    PubMed Central

    Mulvey, Brett B.; Olcese, Ursula; Cabrera, Janel R.; Horabin, Jamila I.

    2014-01-01

    Genome analysis in several eukaryotes shows a surprising number of transcripts which do not encode conventional messenger RNAs. Once considered noise, these non-coding RNAs (ncRNAs) appear capable of controlling gene expression by various means. We find Drosophila sex determination, specifically the master-switch gene Sex-lethal (Sxl), is regulated by long ncRNAs (>200 nt). The lncRNAs influence the dose sensitive establishment promoter of Sxl, SxlPe, which must be activated to specify female sex. They are primarily from two regions, R1 and R2, upstream of SxlPeand show a dynamic developmental profile. Of the four lncRNA strands only one, R2 antisense, has its peak coincident with SxlPe transcription, suggesting it may promote activation. Indeed, its expression is regulated by the X chromosome counting genes, whose dose determines whether SxlPe is transcribed. Transgenic lines which ectopically express each of the lncRNAs show they can act in trans, impacting the process of sex determination but also altering the levels of the other lncRNAs. Generally, expression of R1 is negative whereas R2 is positive to females. This ectopic expression also results in a change in the local chromatin marks, affecting the timing and strength of SxlPe transcription. The chromatin marks are those deposited by the Polycomb and Trithorax groups of chromatin modifying proteins, which we find bind to the lncRNAs. We suggest the increasing numbers of non-coding transcripts being identified are a harbinger of interacting networks similar to the one we describe. PMID:24954180

  9. Median and ulnar nerve conduction determinations in the Erb's point--axilla segment in normal subjects.

    PubMed Central

    Ginzburg, M; Lee, M; Ginzburg, J; Alba, A

    1978-01-01

    Twenty-one median and 22 ulnar nerves were tested in 12 patients for motor nerve conduction velocity (MNCV) and motor nerve conduction time (MNCT) in the segments from Erb's point (N) to axilla (A) bilaterally. It was found that on this segment for both nerves, MNCV values equal to or smaller than 51 m/s or conduction times equal to or longer than 4 ms are to be considered abnormal. For comparative studies and for checking the normality of the tested nerves in their entire length, the more distally located segments in the same nerve were also tested. For diagnostic purposes, the differences between right and left MNCV or MNCT values determined in the same person on N-A segments of homologous nerves were analysed. Motor nerve conduction velocity or MNCT determinations on the N-A nerve segment are expected to replace MNCV determinations on the longer N-AE (AE=100 mm above elbow) nerve segment, which is now in use, for diagnosis of the thoracic outlet syndrome. Images PMID:660207

  10. Determination of gene expression patterns using high-throughput RNA in situ hybridizaion to whole-mount Drosophila embryos

    SciTech Connect

    Weiszmann, R.; Hammonds, A.S.; Celniker, S.E.

    2009-04-09

    We describe a high-throughput protocol for RNA in situ hybridization (ISH) to Drosophila embryos in a 96-well format. cDNA or genomic DNA templates are amplified by PCR and then digoxigenin-labeled ribonucleotides are incorporated into antisense RNA probes by in vitro transcription. The quality of each probe is evaluated before ISH using a RNA probe quantification (dot blot) assay. RNA probes are hybridized to fixed, mixed-staged Drosophila embryos in 96-well plates. The resulting stained embryos can be examined and photographed immediately or stored at 4oC for later analysis. Starting with fixed, staged embryos, the protocol takes 6 d from probe template production through hybridization. Preparation of fixed embryos requires a minimum of 2 weeks to collect embryos representing all stages. The method has been used to determine the expression patterns of over 6,000 genes throughout embryogenesis.

  11. Microevolution of cis-regulatory elements: an example from the pair-rule segmentation gene fushi tarazu in the Drosophila melanogaster subgroup.

    PubMed

    Bakkali, Mohammed

    2011-01-01

    The importance of non-coding DNAs that control transcription is ever noticeable, but the characterization and analysis of the evolution of such DNAs presents challenges not found in the analysis of coding sequences. In this study of the cis-regulatory elements of the pair rule segmentation gene fushi tarazu (ftz) I report the DNA sequences of ftz's zebra element (promoter) and a region containing the proximal enhancer from a total of 45 fly lines belonging to several populations of the species Drosophila melanogaster, D. simulans, D. sechellia, D. mauritiana, D. yakuba, D. teissieri, D. orena and D. erecta. Both elements evolve at slower rate than ftz synonymous sites, thus reflecting their functional importance. The promoter evolves more slowly than the average for ftz's coding sequence while, on average, the enhancer evolves more rapidly, suggesting more functional constraint and effective purifying selection on the former. Comparative analysis of the number and nature of base substitutions failed to detect significant evidence for positive/adaptive selection in transcription-factor-binding sites. These seem to evolve at similar rates to regions not known to bind transcription factors. Although this result reflects the evolutionary flexibility of the transcription factor binding sites, it also suggests a complex and still not completely understood nature of even the characterized cis-regulatory sequences. The latter seem to contain more functional parts than those currently identified, some of which probably transcription factor binding. This study illustrates ways in which functional assignments of sequences within cis-acting sequences can be used in the search for adaptive evolution, but also highlights difficulties in how such functional assignment and analysis can be carried out. PMID:22073317

  12. Myoblast fusion in Drosophila

    SciTech Connect

    Haralalka, Shruti; Abmayr, Susan M.

    2010-11-01

    The body wall musculature of a Drosophila larva is composed of an intricate pattern of 30 segmentally repeated muscle fibers in each abdominal hemisegment. Each muscle fiber has unique spatial and behavioral characteristics that include its location, orientation, epidermal attachment, size and pattern of innervation. Many, if not all, of these properties are dictated by founder cells, which determine the muscle pattern and seed the fusion process. Myofibers are then derived from fusion between a specific founder cell and several fusion competent myoblasts (FCMs) fusing with as few as 3-5 FCMs in the small muscles on the most ventral side of the embryo and as many as 30 FCMs in the larger muscles on the dorsal side of the embryo. The focus of the present review is the formation of the larval muscles in the developing embryo, summarizing the major issues and players in this process. We have attempted to emphasize experimentally-validated details of the mechanism of myoblast fusion and distinguish these from the theoretically possible details that have not yet been confirmed experimentally. We also direct the interested reader to other recent reviews that discuss myoblast fusion in Drosophila, each with their own perspective on the process . With apologies, we use gene nomenclature as specified by Flybase (http://flybase.org) but provide Table 1 with alternative names and references.

  13. Meiosis in male Drosophila

    PubMed Central

    McKee, Bruce D.; Yan, Rihui; Tsai, Jui-He

    2012-01-01

    Meiosis entails sorting and separating both homologous and sister chromatids. The mechanisms for connecting sister chromatids and homologs during meiosis are highly conserved and include specialized forms of the cohesin complex and a tightly regulated homolog synapsis/recombination pathway designed to yield regular crossovers between homologous chromatids. Drosophila male meiosis is of special interest because it dispenses with large segments of the standard meiotic script, particularly recombination, synapsis and the associated structures. Instead, Drosophila relies on a unique protein complex composed of at least two novel proteins, SNM and MNM, to provide stable connections between homologs during meiosis I. Sister chromatid cohesion in Drosophila is mediated by cohesins, ring-shaped complexes that entrap sister chromatids. However, unlike other eukaryotes Drosophila does not rely on the highly conserved Rec8 cohesin in meiosis, but instead utilizes two novel cohesion proteins, ORD and SOLO, which interact with the SMC1/3 cohesin components in providing meiotic cohesion. PMID:23087836

  14. Determination of wing cell fate by the escargot and snail genes in Drosophila.

    PubMed

    Fuse, N; Hirose, S; Hayashi, S

    1996-04-01

    Inset appendages such as the wing and the leg are formed in response to inductive signals in the embryonic field. In Drosophila, cells receiving such signals initiate developmental programs which allow them to become imaginal discs. Subsequently, these discs autonomously organize patterns specific for each appendage. We here report that two related transcription factors, Escargot and Snail that are expressed in the embryonic wing disc, function as intrinsic determinants of the wing cell fate. In escargot or snail mutant embryos, wing-specific expression of Snail, Vestigial and beta-galactosidase regulated by escargot enhancer were found as well as in wild-type embryos. However, in escargot snail double mutant embryos, wing development proceeded until stage 13, but the marker expression was not maintained in later stages, and the invagination of the primordium was absent. From such analyses, it was concluded that Escargot and Snail expression in the wing disc are maintained by their auto- and crossactivation. Ubiquitous escargot or snail expression induced from the hsp70 promoter rescued the escargot snail double mutant phenotype with the effects confined to the prospective wing cells. Similar DNA binding specificities of Escargot and Snail suggest that they control the same set of genes required for wing development. We thus propose the following scenario for early wing disc development. Prospective wing cells respond to the induction by turning on escargot and snail transcription, and become competent for regulation by Escargot and Snail. Such cells initiate auto- and crossregulatory circuits of escargot and snail. The sustained Escargot and Snail expression then activates vestigial and other target genes that are essential for wing development. This maintains the commitment to the wing cell fate and induces wing-specific cell shape change. PMID:8620833

  15. An image segmentation based on a genetic algorithm for determining soil coverage by crop residues.

    PubMed

    Ribeiro, Angela; Ranz, Juan; Burgos-Artizzu, Xavier P; Pajares, Gonzalo; del Arco, Maria J Sanchez; Navarrete, Luis

    2011-01-01

    Determination of the soil coverage by crop residues after ploughing is a fundamental element of Conservation Agriculture. This paper presents the application of genetic algorithms employed during the fine tuning of the segmentation process of a digital image with the aim of automatically quantifying the residue coverage. In other words, the objective is to achieve a segmentation that would permit the discrimination of the texture of the residue so that the output of the segmentation process is a binary image in which residue zones are isolated from the rest. The RGB images used come from a sample of images in which sections of terrain were photographed with a conventional camera positioned in zenith orientation atop a tripod. The images were taken outdoors under uncontrolled lighting conditions. Up to 92% similarity was achieved between the images obtained by the segmentation process proposed in this paper and the templates made by an elaborate manual tracing process. In addition to the proposed segmentation procedure and the fine tuning procedure that was developed, a global quantification of the soil coverage by residues for the sampled area was achieved that differed by only 0.85% from the quantification obtained using template images. Moreover, the proposed method does not depend on the type of residue present in the image. The study was conducted at the experimental farm "El Encín" in Alcalá de Henares (Madrid, Spain). PMID:22163966

  16. An Image Segmentation Based on a Genetic Algorithm for Determining Soil Coverage by Crop Residues

    PubMed Central

    Ribeiro, Angela; Ranz, Juan; Burgos-Artizzu, Xavier P.; Pajares, Gonzalo; Sanchez del Arco, Maria J.; Navarrete, Luis

    2011-01-01

    Determination of the soil coverage by crop residues after ploughing is a fundamental element of Conservation Agriculture. This paper presents the application of genetic algorithms employed during the fine tuning of the segmentation process of a digital image with the aim of automatically quantifying the residue coverage. In other words, the objective is to achieve a segmentation that would permit the discrimination of the texture of the residue so that the output of the segmentation process is a binary image in which residue zones are isolated from the rest. The RGB images used come from a sample of images in which sections of terrain were photographed with a conventional camera positioned in zenith orientation atop a tripod. The images were taken outdoors under uncontrolled lighting conditions. Up to 92% similarity was achieved between the images obtained by the segmentation process proposed in this paper and the templates made by an elaborate manual tracing process. In addition to the proposed segmentation procedure and the fine tuning procedure that was developed, a global quantification of the soil coverage by residues for the sampled area was achieved that differed by only 0.85% from the quantification obtained using template images. Moreover, the proposed method does not depend on the type of residue present in the image. The study was conducted at the experimental farm “El Encín” in Alcalá de Henares (Madrid, Spain). PMID:22163966

  17. The Determination of Relative Elemental Growth Rate Profiles from Segmental Growth Rates (A Methodological Evaluation).

    PubMed Central

    Peters, W. S.; Bernstein, N.

    1997-01-01

    Relative elemental growth rate (REGR) profiles describe spatial patterns of growth intensity; they are indispensable for causal growth analyses. Published methods of REGR profile determination from marking experiments fall in two classes: the profile is either described by a series of segmental growth rates, or calculated as the slope of a function describing the displacement velocities of points along the organ. The latter technique is usually considered superior for theoretical reasons, but to our knowledge, no comparative methodological study of the two approaches is currently available. We formulated a model REGR profile that resembles those reported from primary roots. We established the displacement velocity profile and derived growth trajectories, which enabled us to perform hypothetical marking experiments on the model with varying spacing of marks and durations of measurement. REGR profiles were determined from these data by alternative methods, and results were compared to the original profile. We find that with our model plotting of segmental relative growth rates versus segment position provides exact REGR profile estimations, if the initial segment length is less than 10% of the length of the whole growing zone, and if less than 20% of the growing zone is displaced past its boundary during the measurement. Based on our analysis, we discuss systematic errors that occur in marking experiments. PMID:12223680

  18. Repeat Length and RNA Expression Level Are Not Primary Determinants in CUG Expansion Toxicity in Drosophila Models

    PubMed Central

    Le Mée, Gwenn; Ezzeddine, Nader; Capri, Michèle; Aït-Ahmed, Ounissa

    2008-01-01

    Evidence for an RNA gain-of-function toxicity has now been provided for an increasing number of human pathologies. Myotonic dystrophies (DM) belong to a class of RNA-dominant diseases that result from RNA repeat expansion toxicity. Specifically, DM of type 1 (DM1), is caused by an expansion of CUG repeats in the 3′UTR of the DMPK protein kinase mRNA, while DM of type 2 (DM2) is linked to an expansion of CCUG repeats in an intron of the ZNF9 transcript (ZNF9 encodes a zinc finger protein). In both pathologies the mutant RNA forms nuclear foci. The mechanisms that underlie the RNA pathogenicity seem to be rather complex and not yet completely understood. Here, we describe Drosophila models that might help unravelling the molecular mechanisms of DM1-associated CUG expansion toxicity. We generated transgenic flies that express inducible repeats of different type (CUG or CAG) and length (16, 240, 480 repeats) and then analyzed transgene localization, RNA expression and toxicity as assessed by induced lethality and eye neurodegeneration. The only line that expressed a toxic RNA has a (CTG)240 insertion. Moreover our analysis shows that its level of expression cannot account for its toxicity. In this line, (CTG)240.4, the expansion inserted in the first intron of CG9650, a zinc finger protein encoding gene. Interestingly, CG9650 and (CUG)240.4 expansion RNAs were found in the same nuclear foci. In conclusion, we suggest that the insertion context is the primary determinant for expansion toxicity in Drosophila models. This finding should contribute to the still open debate on the role of the expansions per se in Drosophila and in human pathogenesis of RNA-dominant diseases. PMID:18213375

  19. Insect segmentation: Genes, stripes and segments in "Hoppers".

    PubMed

    French, V

    2001-11-13

    Recent work has revealed that orthologues of several segmentation genes are expressed in the grasshopper embryo, in patterns resembling those shown in Drosophila. This suggests that, despite great differences between the embryos, a hierarchy of gap/pair-rule/segment polarity gene function may be a shared and ancestral feature of insect segmentation. PMID:11719236

  20. Determination of axial rotation angles of limb segments - a new method.

    PubMed

    Cheng, P L; Nicol, A C; Paul, J P

    2000-07-01

    This paper describes a new method of determining the axial rotation angle of a limb segment during three-dimensional movement. Instead of describing the three-dimensional rotation by a three-step rotation (Euler/Cardan angles), a one-step rotation (instantaneous screw axis), or a non-step rotation (floating axis method), the new method uses a two-step rotation to describe the three-dimensional rotation of the limb segment: the rotation of the long axis of the limb segment about a specific axis passing through the proximal joint centre and perpendicular to the long axis of the limb segment, and the axial rotation about the long axis. A short review of previous methods followed by a full description of the principle of the new method with detailed derivation of some important equations (Appendices), comparison with Euler/Cardan angles and a simple experimental demonstration are given in this paper. A method of solving the gimbal-lock problem when using this method is also proposed. PMID:10831758

  1. Genome-wide identification of direct targets of the Drosophila retinal determination protein Eyeless.

    PubMed

    Ostrin, Edwin J; Li, Yumei; Hoffman, Kristi; Liu, Jing; Wang, Keqing; Zhang, Li; Mardon, Graeme; Chen, Rui

    2006-04-01

    The discovery of direct downstream targets of transcription factors (TFs) is necessary for understanding the genetic mechanisms underlying complex, highly regulated processes such as development. In this report, we have used a combinatorial strategy to conduct a genome-wide search for novel direct targets of Eyeless (Ey), a key transcription factor controlling early eye development in Drosophila. To overcome the lack of high-quality consensus binding site sequences, phylogenetic shadowing of known Ey binding sites in sine oculis (so) was used to construct a position weight matrix (PWM) of the Ey protein. This PWM was then used for in silico prediction of potential binding sites in the Drosophila melanogaster genome. To reduce the false positive rate, conservation of these potential binding sites was assessed by comparing the genomic sequences from seven Drosophila species. In parallel, microarray analysis of wild-type versus ectopic ey-expressing tissue, followed by microarray-based epistasis experiments in an atonal (ato) mutant background, identified 188 genes induced by ey. Intersection of in silico predicted conserved Ey binding sites with the candidate gene list produced through expression profiling yields a list of 20 putative ey-induced, eye-enriched, ato-independent, direct targets of Ey. The accuracy of this list of genes was confirmed using both in vitro and in vivo methods. Initial analysis reveals three genes, eyes absent, shifted, and Optix, as novel direct targets of Ey. These results suggest that the integrated strategy of computational biology, genomics, and genetics is a powerful approach to identify direct downstream targets for any transcription factor genome-wide. PMID:16533912

  2. Drosophila Condensin II subunit Chromosome-associated protein D3 regulates cell fate determination through non-cell-autonomous signaling.

    PubMed

    Klebanow, Lindsey R; Peshel, Emanuela C; Schuster, Andrew T; De, Kuntal; Sarvepalli, Kavitha; Lemieux, Madeleine E; Lenoir, Jessica J; Moore, Adrian W; McDonald, Jocelyn A; Longworth, Michelle S

    2016-08-01

    The pattern of the Drosophila melanogaster adult wing is heavily influenced by the expression of proteins that dictate cell fate decisions between intervein and vein during development. dSRF (Blistered) expression in specific regions of the larval wing disc promotes intervein cell fate, whereas EGFR activity promotes vein cell fate. Here, we report that the chromatin-organizing protein CAP-D3 acts to dampen dSRF levels at the anterior/posterior boundary in the larval wing disc, promoting differentiation of cells into the anterior crossvein. CAP-D3 represses KNOT expression in cells immediately adjacent to the anterior/posterior boundary, thus blocking KNOT-mediated repression of EGFR activity and preventing cell death. Maintenance of EGFR activity in these cells depresses dSRF levels in the neighboring anterior crossvein progenitor cells, allowing them to differentiate into vein cells. These findings uncover a novel transcriptional regulatory network influencing Drosophila wing vein development, and are the first to identify a Condensin II subunit as an important regulator of EGFR activity and cell fate determination in vivo. PMID:27317808

  3. Drosophila Condensin II subunit Chromosome-associated protein D3 regulates cell fate determination through non-cell-autonomous signaling

    PubMed Central

    Klebanow, Lindsey R.; Peshel, Emanuela C.; Schuster, Andrew T.; De, Kuntal; Sarvepalli, Kavitha; Lemieux, Madeleine E.; Lenoir, Jessica J.; Moore, Adrian W.; McDonald, Jocelyn A.

    2016-01-01

    The pattern of the Drosophila melanogaster adult wing is heavily influenced by the expression of proteins that dictate cell fate decisions between intervein and vein during development. dSRF (Blistered) expression in specific regions of the larval wing disc promotes intervein cell fate, whereas EGFR activity promotes vein cell fate. Here, we report that the chromatin-organizing protein CAP-D3 acts to dampen dSRF levels at the anterior/posterior boundary in the larval wing disc, promoting differentiation of cells into the anterior crossvein. CAP-D3 represses KNOT expression in cells immediately adjacent to the anterior/posterior boundary, thus blocking KNOT-mediated repression of EGFR activity and preventing cell death. Maintenance of EGFR activity in these cells depresses dSRF levels in the neighboring anterior crossvein progenitor cells, allowing them to differentiate into vein cells. These findings uncover a novel transcriptional regulatory network influencing Drosophila wing vein development, and are the first to identify a Condensin II subunit as an important regulator of EGFR activity and cell fate determination in vivo. PMID:27317808

  4. Cooperative and Antagonistic Contributions of Two Heterochromatin Proteins to Transcriptional Regulation of the Drosophila Sex Determination Decision

    PubMed Central

    Li, Hui; Rodriguez, Janel; Yoo, Youngdong; Shareef, Momin Mohammed; Badugu, RamaKrishna

    2011-01-01

    Eukaryotic nuclei contain regions of differentially staining chromatin (heterochromatin), which remain condensed throughout the cell cycle and are largely transcriptionally silent. RNAi knockdown of the highly conserved heterochromatin protein HP1 in Drosophila was previously shown to preferentially reduce male viability. Here we report a similar phenotype for the telomeric partner of HP1, HOAP, and roles for both proteins in regulating the Drosophila sex determination pathway. Specifically, these proteins regulate the critical decision in this pathway, firing of the establishment promoter of the masterswitch gene, Sex-lethal (Sxl). Female-specific activation of this promoter, SxlPe, is essential to females, as it provides SXL protein to initiate the productive female-specific splicing of later Sxl transcripts, which are transcribed from the maintenance promoter (SxlPm) in both sexes. HOAP mutants show inappropriate SxlPe firing in males and the concomitant inappropriate splicing of SxlPm-derived transcripts, while females show premature firing of SxlPe. HP1 mutants, by contrast, display SxlPm splicing defects in both sexes. Chromatin immunoprecipitation assays show both proteins are associated with SxlPe sequences. In embryos from HP1 mutant mothers and Sxl mutant fathers, female viability and RNA polymerase II recruitment to SxlPe are severely compromised. Our genetic and biochemical assays indicate a repressing activity for HOAP and both activating and repressing roles for HP1 at SxlPe. PMID:21695246

  5. Compositional bias is a major determinant of the distribution pattern and abundance of palindromes in Drosophila melanogaster.

    PubMed

    Liu, Guoqing; Liu, Jia; Zhang, Bingjie

    2012-10-01

    Palindromic sequences are important DNA motifs related to gene regulation, DNA replication and recombination, and thus, investigating the evolutionary forces shaping the distribution pattern and abundance of palindromes in the genome is substantially important. In this article, we analyzed the abundance of palindromes in the genome, and then explored the possible effects of several genomic factors on the palindrome distribution and abundance in Drosophila melanogaster. Our results show that the palindrome abundance in D. melanogaster deviates from random expectation and the uneven distribution of palindromes across the genome is associated with local GC content, recombination rate, and coding exon density. Our data suggest that base composition is the major determinant of the distribution pattern and abundance of palindromes and the correlation between palindrome density and recombination is a side-product of the effect of compositional bias on the palindrome abundance. PMID:23138634

  6. Dscam Expression Levels Determine Presynaptic Arbor Sizes in Drosophila Sensory Neurons

    PubMed Central

    Kim, Jung Hwan; Wang, Xin; Coolon, Rosemary; Ye, Bing

    2013-01-01

    SUMMARY Expression of the Down syndrome cell-adhesion molecule (Dscam) is increased in the brains of patients with several neurological disorders. Although Dscam is critically involved in many aspects of neuronal development, little is known about either the mechanism that regulates its expression or the functional consequences of dysregulated Dscam expression. Here, we show that Dscam expression levels serve as an instructive code for the size control of presynaptic arbor. Two convergent pathways, involving dual leucine zipper kinase (DLK) and fragile X mental retardation protein (FMRP), control Dscam expression through protein translation. Defects in this regulation of Dscam translation lead to exuberant presynaptic arbor growth in Drosophila somatosensory neurons. Our findings demonstrate a previously unknown aspect of Dscam function and provide insights into how dysregulated Dscam may contribute to the pathogenesis of neurological disorders. PMID:23764288

  7. Determination of mechanical stress distribution in Drosophila wing discs using photoelasticity.

    PubMed

    Nienhaus, Ulrike; Aegerter-Wilmsen, Tinri; Aegerter, Christof M

    2009-12-01

    Morphogenesis, the process by which all complex biological structures are formed, is driven by an intricate interplay between genes, growth, as well as intra- and intercellular forces. While the expression of different genes changes the mechanical properties and shapes of cells, growth exerts forces in response to which tissues, organs and more complex structures are shaped. This is exemplified by a number of recent findings for instance in meristem formation in Arabidopsis and tracheal tube formation in Drosophila. However, growth not only generates forces, mechanical forces can also have an effect on growth rates, as is seen in mammalian tissues or bone growth. In fact, mechanical forces can influence the expression levels of patterning genes, allowing control of morphogenesis via mechanical feedback. In order to study the connections between mechanical stress, growth control and morphogenesis, information about the distribution of stress in a tissue is invaluable. Here, we applied stress-birefringence to the wing imaginal disc of Drosophila melanogaster, a commonly used model system for organ growth and patterning, in order to assess the stress distribution present in this tissue. For this purpose, stress-related differences in retardance are measured using a custom-built optical set-up. Applying this method, we found that the stresses are inhomogeneously distributed in the wing disc, with maximum compression in the centre of the wing pouch. This compression increases with wing disc size, showing that mechanical forces vary with the age of the tissue. These results are discussed in light of recent models proposing mechanical regulation of wing disc growth. PMID:19748573

  8. Molecular Determinants of Juvenile Hormone Action as Revealed by 3D QSAR Analysis in Drosophila

    PubMed Central

    Beňo, Milan; Farkaš, Robert

    2009-01-01

    Background Postembryonic development, including metamorphosis, of many animals is under control of hormones. In Drosophila and other insects these developmental transitions are regulated by the coordinate action of two principal hormones, the steroid ecdysone and the sesquiterpenoid juvenile hormone (JH). While the mode of ecdysone action is relatively well understood, the molecular mode of JH action remains elusive. Methodology/Principal Findings To gain more insights into the molecular mechanism of JH action, we have tested the biological activity of 86 structurally diverse JH agonists in Drosophila melanogaster. The results were evaluated using 3D QSAR analyses involving CoMFA and CoMSIA procedures. Using this approach we have generated both computer-aided and species-specific pharmacophore fingerprints of JH and its agonists, which revealed that the most active compounds must possess an electronegative atom (oxygen or nitrogen) at both ends of the molecule. When either of these electronegative atoms are replaced by carbon or the distance between them is shorter than 11.5 Å or longer than 13.5 Å, their biological activity is dramatically decreased. The presence of an electron-deficient moiety in the middle of the JH agonist is also essential for high activity. Conclusions/Significance The information from 3D QSAR provides guidelines and mechanistic scope for identification of steric and electrostatic properties as well as donor and acceptor hydrogen-bonding that are important features of the ligand-binding cavity of a JH target protein. In order to refine the pharmacophore analysis and evaluate the outcomes of the CoMFA and CoMSIA study we used pseudoreceptor modeling software PrGen to generate a putative binding site surrogate that is composed of eight amino acid residues corresponding to the defined molecular interactions. PMID:19547707

  9. Method 349.0 Determination of Ammonia in Estuarine and Coastal Waters by Gas Segmented Continuous Flow Colorimetric Analysis

    EPA Science Inventory

    This method provides a procedure for the determination of ammonia in estuarine and coastal waters. The method is based upon the indophenol reaction,1-5 here adapted to automated gas-segmented continuous flow analysis.

  10. Fuzzy hidden Markov chains segmentation for volume determination and quantitation in PET

    NASA Astrophysics Data System (ADS)

    Hatt, M.; Lamare, F.; Boussion, N.; Turzo, A.; Collet, C.; Salzenstein, F.; Roux, C.; Jarritt, P.; Carson, K.; Cheze-LeRest, C.; Visvikis, D.

    2007-07-01

    Accurate volume of interest (VOI) estimation in PET is crucial in different oncology applications such as response to therapy evaluation and radiotherapy treatment planning. The objective of our study was to evaluate the performance of the proposed algorithm for automatic lesion volume delineation; namely the fuzzy hidden Markov chains (FHMC), with that of current state of the art in clinical practice threshold based techniques. As the classical hidden Markov chain (HMC) algorithm, FHMC takes into account noise, voxel intensity and spatial correlation, in order to classify a voxel as background or functional VOI. However the novelty of the fuzzy model consists of the inclusion of an estimation of imprecision, which should subsequently lead to a better modelling of the 'fuzzy' nature of the object of interest boundaries in emission tomography data. The performance of the algorithms has been assessed on both simulated and acquired datasets of the IEC phantom, covering a large range of spherical lesion sizes (from 10 to 37 mm), contrast ratios (4:1 and 8:1) and image noise levels. Both lesion activity recovery and VOI determination tasks were assessed in reconstructed images using two different voxel sizes (8 mm3 and 64 mm3). In order to account for both the functional volume location and its size, the concept of % classification errors was introduced in the evaluation of volume segmentation using the simulated datasets. Results reveal that FHMC performs substantially better than the threshold based methodology for functional volume determination or activity concentration recovery considering a contrast ratio of 4:1 and lesion sizes of <28 mm. Furthermore differences between classification and volume estimation errors evaluated were smaller for the segmented volumes provided by the FHMC algorithm. Finally, the performance of the automatic algorithms was less susceptible to image noise levels in comparison to the threshold based techniques. The analysis of both

  11. The IMD innate immunity pathway of Drosophila influences somatic sex determination via regulation of the Doa locus.

    PubMed

    Zhao, Yunpo; Cocco, Claudia; Domenichini, Severine; Samson, Marie-Laure; Rabinow, Leonard

    2015-11-15

    The IMD pathway induces the innate immune response to infection by gram-negative bacteria. We demonstrate strong female-to-male sex transformations in double mutants of the IMD pathway in combination with Doa alleles. Doa encodes a protein kinase playing a central role in somatic sex determination through its regulation of alternative splicing of dsx transcripts. Transcripts encoding two specific Doa isoforms are reduced in Rel null mutant females, supporting our genetic observations. A role for the IMD pathway in somatic sex determination is further supported by the induction of female-to-male sex transformations by Dredd mutations in sensitized genetic backgrounds. In contrast, mutations in either dorsal or Dif, the two other NF-κB paralogues of Drosophila, display no effects on sex determination, demonstrating the specificity of IMD signaling. Our results reveal a novel role for the innate immune IMD signaling pathway in the regulation of somatic sex determination in addition to its role in response to microbial infection, demonstrating its effects on alternative splicing through induction of a crucial protein kinase. PMID:26434917

  12. Conditional knockout of retinal determination genes in differentiating cells in Drosophila.

    PubMed

    Jin, Meng; Eblimit, Aiden; Pulikkathara, Merlyn; Corr, Stuart; Chen, Rui; Mardon, Graeme

    2016-08-01

    Conditional gene knockout in postmitotic cells is a valuable technique which allows the study of gene function with spatiotemporal control. Surprisingly, in contrast to its long-term and extensive use in mouse studies, this technology is lacking in Drosophila. Here, we use a novel method for generating complete loss of eyes absent (eya) or sine oculis (so) function in postmitotic cells posterior to the morphogenetic furrow (MF). Specifically, genomic rescue constructs with flippase recognition target (FRT) sequences flanking essential exons are used to generate conditional null alleles. By removing gene function in differentiating cells, we show that eya and so are dispensable for larval photoreceptor differentiation, but are required for differentiation during pupal development. Both eya and so are necessary for photoreceptor survival and the apoptosis caused by loss of eya or so function is likely a secondary consequence of inappropriate differentiation. We also confirm their requirement for cone cell development and reveal a novel role in interommatidial bristle (IOB) formation. In addition, so is required for normal eye disc morphology. This is the first report of a knockout method to study eya and so function in postmitotic cells. This technology will open the door to a large array of new functional studies in virtually any tissue and at any stage of development or in adults. PMID:27257739

  13. Enhancer additivity and non-additivity are determined by enhancer strength in the Drosophila embryo

    PubMed Central

    Bothma, Jacques P; Garcia, Hernan G; Ng, Samuel; Perry, Michael W; Gregor, Thomas; Levine, Michael

    2015-01-01

    Metazoan genes are embedded in a rich milieu of regulatory information that often includes multiple enhancers possessing overlapping activities. In this study, we employ quantitative live imaging methods to assess the function of pairs of primary and shadow enhancers in the regulation of key patterning genes-knirps, hunchback, and snail-in developing Drosophila embryos. The knirps enhancers exhibit additive, sometimes even super-additive activities, consistent with classical gene fusion studies. In contrast, the hunchback enhancers function sub-additively in anterior regions containing saturating levels of the Bicoid activator, but function additively in regions where there are diminishing levels of the Bicoid gradient. Strikingly sub-additive behavior is also observed for snail, whereby removal of the proximal enhancer causes a significant increase in gene expression. Quantitative modeling of enhancer–promoter interactions suggests that weakly active enhancers function additively while strong enhancers behave sub-additively due to competition with the target promoter. DOI: http://dx.doi.org/10.7554/eLife.07956.001 PMID:26267217

  14. Contextual interactions determine whether the Drosophila homeodomain protein, Vnd, acts as a repressor or activator

    PubMed Central

    Yu, Zhongxin; Syu, Li-Jyun; Mellerick, Dervla M.

    2005-01-01

    At the molecular level, members of the NKx2.2 family of transcription factors establish neural compartment boundaries by repressing the expression of homeobox genes specific for adjacent domains [Muhr et al. (2001) Cell, 104, 861–873; Weiss et al. (1998) Genes Dev., 12, 3591–3602]. The Drosophila homologue, vnd, interacts genetically with the high-mobility group protein, Dichaete, in a manner suggesting co-operative activation [Zhao and Skeath (2002) Development, 129, 1165–1174]. However, evidence for direct interactions and transcriptional activation is lacking. Here, we present molecular evidence for the interaction of Vnd and Dichaete that leads to the activation of target gene expression. Two-hybrid interaction assays indicate that Dichaete binds the Vnd homeodomain, and additional Vnd sequences stabilize this interaction. In addition, Vnd has two activation domains that are typically masked in the intact protein. Whether vnd can activate or repress transcription is context-dependent. Full-length Vnd, when expressed as a Gal4 fusion protein, acts as a repressor containing multiple repression domains. A divergent domain in the N-terminus, not found in vertebrate Vnd-like proteins, causes the strongest repression. The co-repressor, Groucho, enhances Vnd repression, and these two proteins physically interact. The data presented indicate that the activation and repression domains of Vnd are complex, and whether Vnd functions as a transcriptional repressor or activator depends on both intra- and inter-molecular interactions. PMID:15640442

  15. Mutations in the white gene of Drosophila melanogaster affecting ABC transporters that determine eye colouration.

    PubMed

    Mackenzie, S M; Brooker, M R; Gill, T R; Cox, G B; Howells, A J; Ewart, G D

    1999-07-15

    The white, brown and scarlet genes of Drosophila melanogaster encode proteins which transport guanine or tryptophan (precursors of the red and brown eye colour pigments) and belong to the ABC transporter superfamily. Current models envisage that the white and brown gene products interact to form a guanine specific transporter, while white and scarlet gene products interact to form a tryptophan transporter. In this study, we report the nucleotide sequence of the coding regions of five white alleles isolated from flies with partially pigmented eyes. In all cases, single amino acid changes were identified, highlighting residues with roles in structure and/or function of the transporters. Mutations in w(cf) (G589E) and w(sat) (F590G) occur at the extracellular end of predicted transmembrane helix 5 and correlate with a major decrease in red pigments in the eyes, while brown pigments are near wild-type levels. Therefore, those residues have a more significant role in the guanine transporter than the tryptophan transporter. Mutations identified in w(crr) (H298N) and w(101) (G243S) affect amino acids which are highly conserved among the ABC transporter superfamily within the nucleotide binding domain. Both cause substantial and similar decreases of red and brown pigments indicating that both tryptophan and guanine transport are impaired. The mutation identified in w(Et87) alters an amino acid within an intracellular loop between transmembrane helices 2 and 3 of the predicted structure. Red and brown pigments are reduced to very low levels by this mutation indicating this loop region is important for the function of both guanine and tryptophan transporters. PMID:10407069

  16. Characterization of the Structural and Functional Determinants of MANF/CDNF in Drosophila In Vivo Model

    PubMed Central

    Lindström, Riitta; Lindholm, Päivi; Kallijärvi, Jukka; Yu, Li-ying; Piepponen, T. Petteri; Arumäe, Urmas; Saarma, Mart; Heino, Tapio I.

    2013-01-01

    Mammalian MANF and CDNF proteins are evolutionarily conserved neurotrophic factors that can protect and repair mammalian dopaminergic neurons in vivo. In Drosophila, the sole MANF protein (DmManf) is needed for the maintenance of dopaminergic neurites and dopamine levels. Although both secreted and intracellular roles for MANF and CDNF have been demonstrated, very little is known about the molecular mechanism of their action. Here, by using a transgenic rescue approach in the DmManf mutant background we show that only full-length MANF containing both the amino-terminal saposin-like and carboxy-terminal SAP-domains can rescue the larval lethality of the DmManf mutant. Independent N- or C-terminal domains of MANF, even when co-expressed together, fail to rescue. Deleting the signal peptide or mutating the CXXC motif in the C-terminal domain destroys the activity of full-length DmManf. Positively charged surface amino acids and the C-terminal endoplasmic reticulum retention signal are necessary for rescue of DmManf mutant lethality when DmManf is expressed in a restricted pattern. Furthermore, rescue experiments with non-ubiquitous expression reveals functional differences between the C-terminal domain of human MANF and CDNF. Finally, DmManf and its C-terminal domain rescue mammalian sympathetic neurons from toxin-induced apoptosis in vitro demonstrating functional similarity of the mammalian and fly proteins. Our study offers further insights into the functional conservation between invertebrate and mammalian MANF/CDNF proteins and reveals the importance of the C-terminal domain for MANF activity in vivo. PMID:24019940

  17. Nemo phosphorylates Eyes absent and enhances output from the Eya-Sine oculis transcriptional complex during Drosophila retinal determination

    PubMed Central

    Morillo, Santiago A.; Braid, Lorena R.; Verheyen, Esther M.; Rebay, Ilaria

    2013-01-01

    The retinal determination gene network comprises a collection of transcription factors that respond to multiple signaling inputs to direct Drosophila eye development. Previous genetic studies have shown that nemo (nmo), a gene encoding a proline-directed serine/threonine kinase, can promote retinal specification through interactions with the retinal determination gene network, although the molecular point of cross-talk was not defined. Here, we report that the Nemo kinase positively and directly regulates Eyes absent (Eya). Genetic assays show that Nmo catalytic activity enhances Eya-mediated ectopic eye formation and potentiates induction of the Eya-Sine oculis (So) transcriptional targets dachshund and lozenge. Biochemical analyses demonstrate that Nmo forms a complex with and phosphorylates Eya at two consensus mitogen-activated protein kinase (MAPK) phosphorylation sites. These same sites appear crucial for Nmo-mediated activation of Eya function in vivo. Thus, we propose that Nmo phosphorylation of Eya potentiates its transactivation function to enhance transcription of Eya-So target genes during eye specification and development. PMID:22394486

  18. The Drosophila melanogaster sex determination gene sisA is required in yolk nuclei for midgut formation.

    PubMed Central

    Walker, J J; Lee, K K; Desai, R N; Erickson, J W

    2000-01-01

    During sex determination, the sisterlessA (sisA) gene functions as one of four X:A numerator elements that set the alternative male or female regulatory states of the switch gene Sex-lethal. In somatic cells, sisA functions specifically in sex determination, but its expression pattern also hints at a role in the yolk cell, a syncytial structure believed to provide energy and nutrients to the developing embryo. Previous studies of sisA have been limited by the lack of a null allele, leaving open the possibility that sisA has additional functions. Here we report the isolation and molecular characterization of four new sisA alleles including two null mutations. Our findings highlight key aspects of sisA structure-function and reveal important qualitative differences between the effects of sisA and the other strong X:A numerator element, sisterlessB, on Sex-lethal expression. We use genetic, expression, clonal, and phenotypic analyses to demonstrate that sisA has an essential function in the yolk nuclei of both sexes. In the absence of sisA, endoderm migration and midgut formation are blocked, suggesting that the yolk cell may have a direct role in larval gut development. To our knowledge, this is the first report of a requirement for the yolk nuclei in Drosophila development. PMID:10790394

  19. Pulse shape analysis and position determination in segmented HPGe detectors: The AGATA detector library

    NASA Astrophysics Data System (ADS)

    Bruyneel, B.; Birkenbach, B.; Reiter, P.

    2016-03-01

    The AGATA Detector Library (ADL) was developed for the calculation of signals from highly segmented large volume high-purity germanium (HPGe) detectors. ADL basis sets comprise a huge amount of calculated position-dependent detector pulse shapes. A basis set is needed for Pulse Shape Analysis (PSA). By means of PSA the interaction position of a γ-ray inside the active detector volume is determined. Theoretical concepts of the calculations are introduced and cover the relevant aspects of signal formation in HPGe. The approximations and the realization of the computer code with its input parameters are explained in detail. ADL is a versatile and modular computer code; new detectors can be implemented in this library. Measured position resolutions of the AGATA detectors based on ADL are discussed.

  20. Isolation of a human gene with protein sequence similarity to human and murine int-1 and the Drosophila segment polarity mutant wingless.

    PubMed Central

    Wainwright, B J; Scambler, P J; Stanier, P; Watson, E K; Bell, G; Wicking, C; Estivill, X; Courtney, M; Boue, A; Pedersen, P S

    1988-01-01

    An expressed gene sequence which was identified by the isolation of a methylation free CpG island from human chromosome 7 has been cloned from a human lung cDNA library. The deduced protein sequence contains 360 amino acids and has several features of a secreted protein; it is cysteine rich with a signal peptide sequence and two potential asn-linked glycosylation sites. The protein sequence shows marked similarity with human and murine int-1 and their Drosophila homolog wingless (Dint-1). This human int-1 related protein, int-1 and Dint-1 have diverse patterns of expression, but the inferred structural similarities suggest that some of the functional characteristics of these proteins may be shared. Images PMID:2971536

  1. Host genetic determinants of microbiota-dependent nutrition revealed by genome-wide analysis of Drosophila melanogaster

    PubMed Central

    Dobson, Adam J.; Chaston, John M.; Newell, Peter D.; Donahue, Leanne; Hermann, Sara L.; Sannino, David R.; Westmiller, Stephanie; Wong, Adam C.-N.; Clark, Andrew G.; Lazzaro, Brian P.; Douglas, Angela E.

    2015-01-01

    Animals bear communities of gut microorganisms with substantial effects on animal nutrition, but the host genetic basis of these effects is unknown. Here, we use Drosophila to demonstrate substantial among-genotype variation in the effects of eliminating the gut microbiota on five host nutritional indices (weight, and protein, lipid, glucose and glycogen contents); this includes variation in both the magnitude and direction of microbiota-dependent effects. Genome-wide associations to identify the genetic basis of the microbiota-dependent variation reveal polymorphisms in largely non-overlapping sets of genes associated with variation in the nutritional traits, including strong representation of conserved genes functioning in signaling. Key genes identified by the GWA study are validated by loss-of-function mutations that altered microbiota-dependent nutritional effects. We conclude that the microbiota interacts with the animal at multiple points in the signaling and regulatory networks that determine animal nutrition. These interactions with the microbiota are likely conserved across animals, including humans. PMID:25692519

  2. distal antenna and distal antenna-related function in the retinal determination network during eye development in Drosophila

    PubMed Central

    Curtiss, Jennifer; Burnett, Micheal; Mlodzik, Marek

    2007-01-01

    Drosophila eye specification occurs through the activity of the retinal determination (RD) network, which includes the Eyeless (Ey), Eyes absent (Eya), Sine oculis (So) and Dachshund (Dac) transcription factors. Based on their abilities to transform antennal precursors towards an eye fate, the distal antenna (dan) and distal antenna-related (danr) genes encode two new RD factors. Dan and Danr are probable transcription factors localized in nuclei of eye precursors and differentiating eye tissue. Loss-of-function single and double dan/danr mutants have small, rough eyes, indicating a requirement for wild-type eye development. In addition, dan and danr participate in the transcriptional hierarchy that controls expression of RD genes, and Dan and Danr interact physically and genetically with Ey and Dac. Eye specification culminates in differentiation of ommatidia through the activities of the proneural gene atonal (ato) in the founding R8 photoreceptor and Egfr signaling in additional photoreceptors. Danr expression overlaps with Ato during R8 specification, and Dan and Danr regulate Ato expression and are required for normal R8 induction and differentiation. These data demonstrate a role for Dan and Danr in eye development and provide a link between eye specification and differentiation. PMID:17493605

  3. Interplay of dFOXO and Two ETS-Family Transcription Factors Determines Lifespan in Drosophila melanogaster

    PubMed Central

    Alic, Nazif; Giannakou, Maria E.; Papatheodorou, Irene; Hoddinott, Matthew P.; Andrews, T. Daniel; Bolukbasi, Ekin; Partridge, Linda

    2014-01-01

    Forkhead box O (FoxO) transcription factors (TFs) are key drivers of complex transcriptional programmes that determine animal lifespan. FoxOs regulate a number of other TFs, but how these TFs in turn might mediate the anti-ageing programmes orchestrated by FoxOs in vivo is unclear. Here, we identify an E-twenty six (ETS)-family transcriptional repressor, Anterior open (Aop), as regulated by the single Drosophila melanogaster FoxO (dFOXO) in the adult gut. AOP, the functional orthologue of the human Etv6/Tel protein, binds numerous genomic sites also occupied by dFOXO and counteracts the activity of an ETS activator, Pointed (Pnt), to prevent the lifespan-shortening effects of co-activation of dFOXO and PNT. This detrimental synergistic effect of dFOXO and PNT appears to stem from a mis-regulation of lipid metabolism. At the same time, AOP activity in another fly organ, the fat body, has further beneficial roles, regulating genes in common with dfoxo, such as the secreted, non-sensory, odorant binding protein (Obp99b), and robustly extending lifespan. Our study reveals a complex interplay between evolutionarily conserved ETS factors and dFOXO, the functional significance of which may extend well beyond animal lifespan. PMID:25232726

  4. Determination of the temperature-sensitive period of a new mutation lawc{sup P1} in Drosophila melanogaster

    SciTech Connect

    Simonova, O.B.; Petruk, S.F.; Gerasimova, T.I.

    1995-09-01

    Determination of the period of temperature sensitivity in the temperature-sensitive allele of the regulatory lawc{sup P1} mutation was performed. Homeotic transformation of arista into tarsus, frequency of leg deformation, and bristle superexpression were examined. The sensitive periods were detected with reciprocal changes of cultivation temperature from 28 to 17{degrees}C and from 17 to 28{degrees}C. The temperature-sensitive period (TSP) for arista transformation was shown to manifest polyphasic expression and sexual dimorphism. In females, it occurred in the late third instar larvae (the first phase) and prepupae (the second stage); in males, it includes the whole period from the late third instar larvae up to and including prepupa. TSP for the frequency of deformed legs was polyphasic and took place during the third larval instar (the first phase) and prepupa stage (the second one). TSP for bristle superexpression occurred during a single interval from the late third larval instar until the early prepupa stage. The products of the lawc gene are assumed to play a role both in the cell proliferation in the antennal and leg imaginal discs and in the control of bristle expression at the final stages of Drosophila ontogeny. 10 refs., 7 figs.

  5. Genetic and molecular analysis of the autosomal component of the primary sex determination signal of Drosophila melanogaster

    SciTech Connect

    Barbash, D.A.; Cline, T.W.

    1995-12-01

    Drosophila sex is determined by the action of the X:A chromosome balance on transcription of Sex-lethal (Sxl), a feminizing switch gene. We obtained loss-of-function mutations in denominator elements of the X:A signal by selecting for dominant suppressors of a female-specific lethal mutation in the numerator element, sisterlessA (sisA). Ten suppressors were recovered in this extensive genome-wide selection. All were mutations in deadpan (dpn), a pleiotropic locus previously discovered to be a denominator element. Detailed genetic and molecular characterization is presented of this diverse set of new dpn alleles including their effects on Sxl. Although selected only for impairment of sex-specific functions, all were also impaired in nonsex-specific functions. Male-lethal effects were anticipated for mutations in a major denominator element, but we found that viability of males lacking dpn function was reduced no more than 50% relative to their dpn{sup -} sisters. Moreover, loss of dpn activity in males caused only a modest depression of the Sxl {open_quotes}establishment{close_quotes} promoter (Sxl{sub Pe}), the X:A target. By itself, dpn cannot account for the masculinizing effect of increased autosomal ploidy, the effect that gave rise to the concept of the X:A ratio; nevertheless, if there are other denominator elements, our results suggest that their individual contributions to the sex-determination signal are even less than that of dpn. The time course of expression of dpn and Sxl in dpn mutant backgrounds suggests that dpn is required for sex determination only during the later stages of X:A signaling in males to prevent inappropriate expression of Sxl{sub Pe} in the face of increasing sis gene product levels. 77 refs., 6 figs., 6 tabs.

  6. Genetic and Molecular Analysis of the Autosomal Component of the Primary Sex Determination Signal of Drosophila Melanogaster

    PubMed Central

    Barbash, D. A.; Cline, T. W.

    1995-01-01

    Drosophila sex is determined by the action of the X:A chromosome balance on transcription of Sex-lethal (Sxl), a feminizing switch gene. We obtained loss-of-function mutations in denominator elements of the X:A signal by selecting for dominant suppressors of a female-specific lethal mutation in the numerator element, sisterlessA (sisA). Ten suppressors were recovered in this extensive genome-wide selection. All were mutations in deadpan (dpn), a pleiotropic locus previously discovered to be a denominator element. Detailed genetic and molecular characterization is presented of this diverse set of new dpn alleles including their effects on Sxl. Although selected only for impairment of sex-specific functions, all were also impaired in nonsex-specific functions. Male-lethal effects were anticipated for mutations in a major denominator element, but we found that viability of males lacking dpn function was reduced no more than 50% relative to their dpn(-) sisters. Moreover, loss of dpn activity in males caused only a modest derepression of the Sxl ``establishment'' promoter (Sxl(Pe)), the X:A target. By itself, dpn cannot account for the masculinizing effect of increased autosomal ploidy, the effect that gave rise to the concept of the X:A ratio; nevertheless, if there are other denominator elements, our results suggest that their individual contributions to the sex-determination signal are even less than that of dpn. The time course of expression of dpn and of Sxl in dpn mutant backgrounds suggests that dpn is required for sex determination only during the later stages of X:A signaling in males to prevent inappropriate expression of Sxl(Pe) in the face of increasing sis gene product levels. PMID:8601486

  7. Contact-free determination of human body segment parameters by means of videometric image processing of an anthropomorphic body model

    NASA Astrophysics Data System (ADS)

    Hatze, Herbert; Baca, Arnold

    1993-01-01

    The development of noninvasive techniques for the determination of biomechanical body segment parameters (volumes, masses, the three principal moments of inertia, the three local coordinates of the segmental mass centers, etc.) receives increasing attention from the medical sciences (e,.g., orthopaedic gait analysis), bioengineering, sport biomechanics, and the various space programs. In the present paper, a novel method is presented for determining body segment parameters rapidly and accurately. It is based on the video-image processing of four different body configurations and a finite mass-element human body model. The four video images of the subject in question are recorded against a black background, thus permitting the application of shape recognition procedures incorporating edge detection and calibration algorithms. In this way, a total of 181 object space dimensions of the subject's body segments can be reconstructed and used as anthropometric input data for the mathematical finite mass- element body model. The latter comprises 17 segments (abdomino-thoracic, head-neck, shoulders, upper arms, forearms, hands, abdomino-pelvic, thighs, lower legs, feet) and enables the user to compute all the required segment parameters for each of the 17 segments by means of the associated computer program. The hardware requirements are an IBM- compatible PC (1 MB memory) operating under MS-DOS or PC-DOS (Version 3.1 onwards) and incorporating a VGA-board with a feature connector for connecting it to a super video windows framegrabber board for which there must be available a 16-bit large slot. In addition, a VGA-monitor (50 - 70 Hz, horizontal scan rate at least 31.5 kHz), a common video camera and recorder, and a simple rectangular calibration frame are required. The advantage of the new method lies in its ease of application, its comparatively high accuracy, and in the rapid availability of the body segment parameters, which is particularly useful in clinical practice

  8. Structural determinants of species-selective substrate recognition in human and Drosophila serotonin transporters revealed through computational docking studies.

    PubMed

    Kaufmann, Kristian W; Dawson, Eric S; Henry, L Keith; Field, Julie R; Blakely, Randy D; Meiler, Jens

    2009-02-15

    To identify potential determinants of substrate selectivity in serotonin (5-HT) transporters (SERT), models of human and Drosophila serotonin transporters (hSERT, dSERT) were built based on the leucine transporter (LeuT(Aa)) structure reported by Yamashita et al. (Nature 2005;437:215-223), PBDID 2A65. Although the overall amino acid identity between SERTs and the LeuT(Aa) is only 17%, it increases to above 50% in the first shell of the putative 5-HT binding site, allowing de novo computational docking of tryptamine derivatives in atomic detail. Comparison of hSERT and dSERT complexed with substrates pinpoints likely structural determinants for substrate binding. Forgoing the use of experimental transport and binding data of tryptamine derivatives for construction of these models enables us to critically assess and validate their predictive power: A single 5-HT binding mode was identified that retains the amine placement observed in the LeuT(Aa) structure, matches site-directed mutagenesis and substituted cysteine accessibility method (SCAM) data, complies with support vector machine derived relations activity relations, and predicts computational binding energies for 5-HT analogs with a significant correlation coefficient (R = 0.72). This binding mode places 5-HT deep in the binding pocket of the SERT with the 5-position near residue hSERT A169/dSERT D164 in transmembrane helix 3, the indole nitrogen next to residue Y176/Y171, and the ethylamine tail under residues F335/F327 and S336/S328 within 4 A of residue D98. Our studies identify a number of potential contacts whose contribution to substrate binding and transport was previously unsuspected. PMID:18704946

  9. Structural determinants of species-selective substrate recognition in human and Drosophila serotonin transporters revealed through computational docking studies

    PubMed Central

    Kaufmann, Kristian W.; Dawson, Eric S.; Henry, L. Keith; Field, Julie R.; Blakely, Randy D.; Meiler, Jens

    2009-01-01

    To identify potential determinants of substrate selectivity in serotonin (5-HT) transporters (SERT), models of human and Drosophila serotonin transporters (hSERT, dSERT) were built based on the leucine transporter (LeuTAa) structure reported by Yamashita et al. (Nature 2005;437:215–223), PBDID 2A65. Although the overall amino acid identity between SERTs and the LeuTAa is only 17%, it increases to above 50% in the first shell of the putative 5-HT binding site, allowing de novo computational docking of tryptamine derivatives in atomic detail. Comparison of hSERT and dSERT complexed with substrates pinpoints likely structural determinants for substrate binding. Forgoing the use of experimental transport and binding data of tryptamine derivatives for construction of these models enables us to cHitically assess and validate their predictive power: A single 5-HT binding mode was identified that retains the amine placement observed in the LeuTAa structure, matches site-directed mutagenesis and substituted cysteine accessibility method (SCAM) data, complies with support vector machine derived relations activity relations, and predicts computational binding energies for 5-HT analogs with a significant correlation coefficient (R = 0.72). This binding mode places 5-HT deep in the binding pocket of the SERT with the 5-position near residue hSERT A169/dSERT D164 in transmembrane helix 3, the indole nitrogen next to residue Y176/Y171, and the ethylamine tail under residues F335/F327 and S336/S328 within 4 Å of residue D98. Our studies identify a number of potential contacts whose contribution to substrate binding and transport was previously unsuspected. PMID:18704946

  10. Recruitment of the proneural gene scute to the Drosophila sex-determination pathway.

    PubMed Central

    Wrischnik, Lisa A; Timmer, John R; Megna, Lisa A; Cline, Thomas W

    2003-01-01

    In flies, scute (sc) works with its paralogs in the achaete-scute-complex (ASC) to direct neuronal development. However, in the family Drosophilidae, sc also acquired a role in the primary event of sex determination, X chromosome counting, by becoming an X chromosome signal element (XSE)-an evolutionary step shown here to have occurred after sc diverged from its closest paralog, achaete (ac). Two temperature-sensitive alleles, sc(sisB2) and sc(sisB3), which disrupt only sex determination, were recovered in a powerful F1 genetic selection and used to investigate how sc was recruited to the sex-determination pathway. sc(sisB2) revealed 3' nontranscribed regulatory sequences likely to be involved. The sc(sisB2) lesion abolished XSE activity when combined with mutations engineered in a sequence upstream of all XSEs. In contrast, changes in Sc protein sequence seem not to have been important for recruitment. The observation that the other new allele, sc(sisB3), eliminates the C-terminal half of Sc without affecting neurogenesis and that sc(sisB1), the most XSE-specific allele previously available, is a nonsense mutant, would seem to suggest the opposite, but we show that housefly Sc can substitute for fruit fly Sc in sex determination, despite lacking Drosophilidae-specific conserved residues in its C-terminal half. Lack of synergistic lethality among mutations in sc, twist, and dorsal argue against a proposed role for sc in mesoderm formation that had seemed potentially relevant to sex-pathway recruitment. The screen that yielded new sc alleles also generated autosomal duplications that argue against the textbook view that fruit fly sex signal evolution recruited a set of autosomal signal elements comparable to the XSEs. PMID:14704182

  11. BRADOS - Dose determination in the Russian segment of the International Space Station

    NASA Astrophysics Data System (ADS)

    Hajek, M.; Berger, T.; Fürstner, M.; Fugger, M.; Vana, N.; Akatov, Y.; Shurshakov, V.; Arkhangelsky, V.

    Absorbed dose and dose-average linear energy transfer (LET) were assessed by means of LiF: Mg, Ti thermoluminescence (TL) detectors at different locations onboard the Russian segment (RS) of the International Space Station (ISS) in the timeframe between February and November 2001, i.e. for 248 days. Based on calibrations of the employed detectors in a variety of heavy-ion beams, mainly at the Heavy Ion Medical Accelerator (HIMAC) in Chiba, Japan, the measured absorbed dose values could be corrected for the TL dose registration efficiency in the radiation climate onboard the ISS. Various strategies for efficiency correction are discussed. For the specific case the efficiency correction accounted for a reduction by nearly 20 % in dose, implying that without proper consideration of the TL efficiency behaviour the absorbed dose inside the ISS would be overestimated. The dose-average LET was derived from TLD-700 measurements evaluated according to the well-established high-temperature ratio (HTR) method which analyzes the TL emission in the temperature range between 248 and 310 C. According to the shielding distribution, the efficiency-corrected absorbed dose was found to vary between 155 μ Gy/d for panel N 457 (RS-ISS toilet) and 230 μ Gy/d for panel N 443 (RS-ISS starboard cabin). The determined LET indicated a modification of the spectral composition of the onboard radiation field for the different exposure locations. Arrangement of TLD-600 and TLD-700 in pair allowed also some information about the neutron component to be drawn. Experimentally determined absorbed dose values are compared with model calculations by means of a self-developed code, using as input data detailed shielding distributions and proton fluxes from AP-8 and JPL algorithms.

  12. Topogenesis of a nucleolar protein: determination of molecular segments directing nucleolar association.

    PubMed

    Zirwes, R F; Kouzmenko, A P; Peters, J M; Franke, W W; Schmidt-Zachmann, M S

    1997-02-01

    To identify the element(s) in nucleolar proteins which determine nucleolus-specific topogenesis, we have used different kinds of cDNA constructs encoding various chimeric combinations of mutants of the constitutive nucleolar protein NO38 (B23): 1) with an amino terminally placed short "myc tag"; 2) with two different carboxyl terminally attached large alpha-helical coiled coil structures, the lamin A rod domain or the rod domain of vimentin; 3) with the sequence-related nucleoplasmic histone-binding protein nucleo-plasmin; and 4) with the soluble cytoplasmic protein pyruvate kinase. To avoid the problem of formation of complexes with endogenous wild-type (wt) molecules and "piggyback" localization, special care was taken to secure that the mutants and chimeras used did not oligomerize as is typical of protein NO38 (B23). Using microinjection and transfection of cultured cells, we found that the segment comprising the amino-terminal 123 amino acids (aa) alone was sufficient to effect nucleolar accumulation of the construct molecules, including the chimeras with the entire rod domains of lamin A and vimentin. However, when the amino-terminal 109 aa were deleted, the molecules still associated with the nucleolus. The results of further deletion experiments and of domain swaps with nucleoplasmin all point to the topogenic importance of two independent molecular regions located at both the amino- and carboxyl-terminal end. Our definition of dominant elements determining the nucleolar localization of protein NO38 (B23) as well as of diverse nonnucleolar proteins will help to identify its local binding partner(s) and functions, the construction of probes examining other proteins or sequence elements within the nucleolar microenvironment, and the generation of cells with an altered nuclear architecture. PMID:9190204

  13. Molecular genetic dissection of the sex-specific and vital functions of the Drosophila melanogaster sex determination gene fruitless.

    PubMed Central

    Anand, A; Villella, A; Ryner, L C; Carlo, T; Goodwin, S F; Song, H J; Gailey, D A; Morales, A; Hall, J C; Baker, B S; Taylor, B J

    2001-01-01

    A multibranched hierarchy of regulatory genes controls all aspects of somatic sexual development in Drosophila melanogaster. One branch of this hierarchy is headed by the fruitless (fru) gene and functions in the central nervous system, where it is necessary for male courtship behavior as well as the differentiation of a male-specific abdominal structure, the muscle of Lawrence (MOL). A preliminary investigation of several of the mutations described here showed that the fru gene also has a sex-nonspecific vital function. The fru gene produces a complex set of transcripts through the use of four promoters and alternative splicing. Only the primary transcripts produced from the most distal (P1) promoter are sex-specifically spliced under direction of the sex-determination hierarchy. We have analyzed eight new fru mutations, created by X-ray mutagenesis and P-element excision, to try to gain insight into the relationship of specific transcript classes to specific fru functions. Males that lack the P1-derived fru transcripts show a complete absence of sexual behavior, but no other defects besides the loss of the MOL. Both males and females that have reduced levels of transcripts from the P3 promoter develop into adults but frequently die after failing to eclose. Analysis of the morphology and behavior of adult escapers showed that P3-encoded functions are required for the proper differentiation and eversion of imaginal discs. Furthermore, the reduction in the size of the neuromuscular junctions on abdominal muscles in these animals suggests that one of fru's sex-nonspecific functions involves general aspects of neuronal differentiation. In mutants that lack all fru transcripts as well as a small number of adjacent genes, animals die at an early pupal stage, indicating that fru's function is required only during late development. Thus, fru functions both in the sex-determination regulatory hierarchy to control male sexual behavior through sex-specific transcripts and

  14. Sex determination in the Drosophila germline is dictated by the sexual identity of the surrounding soma.

    PubMed Central

    Waterbury, J A; Horabin, J I; Bopp, D; Schedl, P

    2000-01-01

    It has been suggested that sexual identity in the germline depends upon the combination of a nonautonomous somatic signaling pathway and an autonomous X chromosome counting system. In the studies reported here, we have examined the role of the sexual differentiation genes transformer (tra) and doublesex (dsx) in regulating the activity of the somatic signaling pathway. We asked whether ectopic somatic expression of the female products of the tra and dsx genes could feminize the germline of XY animals. We find that Tra(F) is sufficient to feminize XY germ cells, shutting off the expression of male-specific markers and activating the expression of female-specific markers. Feminization of the germline depends upon the constitutively expressed transformer-2 (tra-2) gene, but does not seem to require a functional dsx gene. However, feminization of XY germ cells by Tra(F) can be blocked by the male form of the Dsx protein (Dsx(M)). Expression of the female form of dsx, Dsx(F), in XY animals also induced germline expression of female markers. Taken together with a previous analysis of the effects of mutations in tra, tra-2, and dsx on the feminization of XX germ cells in XX animals, our findings indicate that the somatic signaling pathway is redundant at the level tra and dsx. Finally, our studies call into question the idea that a cell-autonomous X chromosome counting system plays a central role in germline sex determination. PMID:10924471

  15. Regulation of the retinal determination gene dachshund in the embryonic head and developing eye of Drosophila

    PubMed Central

    Anderson, Jason; Salzer, Claire L.; Kumar, Justin P.

    2009-01-01

    The retinal determination gene dachshund is distantly related to the family of Ski/Sno proto-oncogenes and influences the development of a wide range of tissues including the embryonic head, optic lobes, brain, central nervous system as well as the post-embryonic leg, wing, genital and eye-antennal discs. We were interested in the regulatory mechanisms that control the dynamic expression pattern of dachshund and in this report we set out to ascertain how the transcription of dachshund is modulated in the embryonic head and developing eye-antennal imaginal disc. We demonstrate that the TGFβ signaling cascade, the transcription factor zerknullt and several other patterning genes prevent dachshund from being expressed inappropriately within the embryonic head. Additionally, we show that several members of the eye specification cascade influence the transcription of dachshund during normal and ectopic eye development. Our results suggest that dachshund is regulated by a complex combinatorial code of transcription factors and signaling pathways. Unraveling this code may lead to an understanding of how dachshund regulates the development of many diverse tissue types including the eye. PMID:16780828

  16. The mesoderm determinant snail collaborates with related zinc-finger proteins to control Drosophila neurogenesis.

    PubMed

    Ashraf, S I; Hu, X; Roote, J; Ip, Y T

    1999-11-15

    The Snail protein functions as a transcriptional regulator to establish early mesodermal cell fate. Later, in germ band-extended embryos, Snail is also expressed in most neuroblasts. Here we present evidence that this expression of Snail is required for central nervous system (CNS) development. The neural function of snail is masked by two closely linked genes, escargot and worniu. Both Escargot and Worniu contain zinc-finger domains that are highly homologous to that of Snail. Although not affecting expression of early neuroblast markers, the deletion of the region containing all three genes correlates with loss of expression of CNS determinants including fushi tarazu, pdm-2 and even-skipped. Transgenic expression of each of the three Snail family proteins can rescue efficiently the fushi tarazu defects, and partially the pdm-2 and even-skipped CNS patterns. These results demonstrate that the Snail family proteins have essential functions during embryonic CNS development, around the time of ganglion mother cell formation. PMID:10562554

  17. Circular DNA Molecules in the Genus Drosophila

    PubMed Central

    Travaglini, E. C.; Schultz, J.

    1972-01-01

    The satellite DNA's from the embryos of five species of Drosophila (D. melanogaster, D. simulans, D. nasuta, D. virilis and D. hydei) have been analyzed for the presence of closed circular duplex DNA molecules, as determined by CsCl-EBr gradients. Circular DNA molecules were found in every species but D. melanogaster. Analyses of cell fractions from adult Drosophila and organ fractions from Drosophila larvae show that fractions containing mitochondria are highly enriched in these molecules. PMID:4643820

  18. Do HLA genes play a prominent role in determining T cell receptor V{alpha} segment usage in humans?

    SciTech Connect

    Gulwani-Akolkar, B.; Shi, B.; Akolkar, P.N.

    1995-04-15

    Previous studies in humans have demonstrated that HLA genes can profoundly influence the TCR V{beta} repertoire. To similarly assess the influence of HLA genes on the TCR V{alpha} segment repertoire, the V{alpha} repertoires of 12 individuals from three unrelated families were determined by quantitative PCR. Each family contained at least one pair of HLA-identical and -nonidentical siblings. Repertoire analysis was performed on purified CD4{sup +} and CD8{sup +} cells by using V{alpha}-specific primers. We were unable to demonstrate more similar V{alpha} repertoires between HLA-identical siblings than between HLA-nonidentical siblings. In contrast, when a similar analysis was performed on the same individuals for the V{beta} repertoire, HLA-identical siblings were found to have significantly more similar repertoires than HLA-nonidentical siblings. Furthermore, both the V{alpha} and V{beta} repertoires of monozygotic twins showed striking similarity. Despite our inability to shown an influence of HLA genes on the V{alpha} repertoire, we did observe a very strong skewing in terms of preferential expression on CD4{sup +} or CD8{sup +} cells of several V{alpha} segments, notably TCRAV1, -2, -5, -6, -7, -11, -12, and -13. These studies suggest that HLA genes play less of a role in determining V{alpha} segment usage than V{beta}. Nevertheless, the pronounced skewing of V{alpha} segment expression in the CD4{sup +} or CD8{sup +} populations suggests some role for HLA genes in determining the V{alpha} TCR repertoire. Furthermore, the striking similarity of V{alpha} repertoires of identical twins suggests a major role for non-HLA genes in determining the V{alpha} repertoire. 35 refs., 8 figs., 3 tabs.

  19. Identification of regions interacting with ovo{sup D} mutations: Potential new genes involved in germline sex determination or differentiation in Drosophila melanogaster

    SciTech Connect

    Pauli, D.; Oliver, B.; Mahowald, A.P.

    1995-02-01

    Only a few Drosophila melanogaster germline sex determination genes are known, and there have been no systematic screens to identify new genes involved in this important biological process. The ovarian phenotypes produced by females mutant for dominant alleles of the ovo gene are modified in flies with altered doses of other loci involved in germline sex determination in Drosophila (Sex-lethal{sup +}, snas fille{sup +} and ovarian tumor{sup +}). This observation constitutes the basis for a screen to identify additional genes required for proper establishment of germline sexual identity. We tested 300 deletions, which together cover {approximately}58% of the euchromatic portion of the genome, for genetic interactions with ovo{sup D}. Hemizygosity for more than a dozen small regions show interactions that either partially suppress or enhance the ovarian phenotypes of females mutant for one or more of the three dominant ovo mutations. These regions probably contain genes whose products act in developmental heirarchies that include ovo{sup +} protein. 40 refs, 7 figs., 5 tabs.

  20. A Circadian Clock Gene, Cry, Affects Heart Morphogenesis and Function in Drosophila as Revealed by Optical Coherence Microscopy

    PubMed Central

    Zeng, Xianxu; Tate, Rebecca E.; McKee, Mary L.; Capen, Diane E.; Zhang, Zhan; Tanzi, Rudolph E.; Zhou, Chao

    2015-01-01

    Circadian rhythms are endogenous, entrainable oscillations of physical, mental and behavioural processes in response to local environmental cues such as daylight, which are present in the living beings, including humans. Circadian rhythms have been related to cardiovascular function and pathology. However, the role that circadian clock genes play in heart development and function in a whole animal in vivo are poorly understood. The Drosophila cryptochrome (dCry) is a circadian clock gene that encodes a major component of the circadian clock negative feedback loop. Compared to the embryonic stage, the relative expression levels of dCry showed a significant increase (>100-fold) in Drosophila during the pupa and adult stages. In this study, we utilized an ultrahigh resolution optical coherence microscopy (OCM) system to perform non-invasive and longitudinal analysis of functional and morphological changes in the Drosophila heart throughout its post-embryonic lifecycle for the first time. The Drosophila heart exhibited major morphological and functional alterations during its development. Notably, heart rate (HR) and cardiac activity period (CAP) of Drosophila showed significant variations during the pupa stage, when heart remodeling took place. From the M-mode (2D + time) OCM images, cardiac structural and functional parameters of Drosophila at different developmental stages were quantitatively determined. In order to study the functional role of dCry on Drosophila heart development, we silenced dCry by RNAi in the Drosophila heart and mesoderm, and quantitatively measured heart morphology and function in those flies throughout its development. Silencing of dCry resulted in slower HR, reduced CAP, smaller heart chamber size, pupal lethality and disrupted posterior segmentation that was related to increased expression of a posterior compartment protein, wingless. Collectively, our studies provided novel evidence that the circadian clock gene, dCry, plays an essential

  1. A Circadian Clock Gene, Cry, Affects Heart Morphogenesis and Function in Drosophila as Revealed by Optical Coherence Microscopy.

    PubMed

    Alex, Aneesh; Li, Airong; Zeng, Xianxu; Tate, Rebecca E; McKee, Mary L; Capen, Diane E; Zhang, Zhan; Tanzi, Rudolph E; Zhou, Chao

    2015-01-01

    Circadian rhythms are endogenous, entrainable oscillations of physical, mental and behavioural processes in response to local environmental cues such as daylight, which are present in the living beings, including humans. Circadian rhythms have been related to cardiovascular function and pathology. However, the role that circadian clock genes play in heart development and function in a whole animal in vivo are poorly understood. The Drosophila cryptochrome (dCry) is a circadian clock gene that encodes a major component of the circadian clock negative feedback loop. Compared to the embryonic stage, the relative expression levels of dCry showed a significant increase (>100-fold) in Drosophila during the pupa and adult stages. In this study, we utilized an ultrahigh resolution optical coherence microscopy (OCM) system to perform non-invasive and longitudinal analysis of functional and morphological changes in the Drosophila heart throughout its post-embryonic lifecycle for the first time. The Drosophila heart exhibited major morphological and functional alterations during its development. Notably, heart rate (HR) and cardiac activity period (CAP) of Drosophila showed significant variations during the pupa stage, when heart remodeling took place. From the M-mode (2D + time) OCM images, cardiac structural and functional parameters of Drosophila at different developmental stages were quantitatively determined. In order to study the functional role of dCry on Drosophila heart development, we silenced dCry by RNAi in the Drosophila heart and mesoderm, and quantitatively measured heart morphology and function in those flies throughout its development. Silencing of dCry resulted in slower HR, reduced CAP, smaller heart chamber size, pupal lethality and disrupted posterior segmentation that was related to increased expression of a posterior compartment protein, wingless. Collectively, our studies provided novel evidence that the circadian clock gene, dCry, plays an essential

  2. Determination of shower central position in laterally segmented lead-fluoride electromagnetic calorimeters

    NASA Astrophysics Data System (ADS)

    Mazouz, M.; Ghedira, L.; Voutier, E.

    2016-07-01

    The spatial resolution of laterally segmented electromagnetic calorimeters, built of lead fluoride material, is studied on the basis of Monte-Carlo simulations. Parametrization of the relative resolution on the shower position is proposed and optimized in terms of the energy of incoming particles and the elementary size of the calorimeter blocks. A new fit algorithm method is proposed that improves spatial resolution at high energies (> 5 GeV), and provides guidance for the design optimization of electromagnetic calorimeters.

  3. Fast Determination of the Planar Body Segment Inertial Parameters Using Affordable Sensors.

    PubMed

    Bonnet, Vincent; Venture, Gentiane

    2015-07-01

    This study aimed at developing and evaluating a new method for the fast and reliable identification of body segment inertial parameters with a planar model using affordable sensors. A Kinect sensor, with a new marker-based tracking system, and a Wii balance board were used as an affordable and portable motion capture system. A set of optimal exciting motions was used in a biofeedback interface to identify the body segment parameters. The method was validated with 12 subjects performing various standardized motions. The same dynamometric quantities estimated both with the proposed system and, as a reference, with a laboratory grade force-plate were compared. The results showed that the proposed method could successfully estimate the resultant moment and the vertical ground reaction force (rms errors less than 8 Nm and 12 N, respectively). Finally, when local segment values were artificially varied, the proposed method was able to detect and estimate the additional masses accurately and with an error of less than 0.5 Kg, contrary to values generated with commonly used anthropometric tables. PMID:25751867

  4. Comparison of automated segmented-flow and discrete analyzers for the determination of nutrients in water

    USGS Publications Warehouse

    Marti, V.C.; Hale, D.R.

    1981-01-01

    Water samples with specific conductances ranging from 66 to 6950 ??mho/cm at 25 ??C were analyzed for ammonia-N (NH3-N), nitrate plus nitrite-N (NO3 + NO2-N), nitrite-N (NO2-N), and phosphate-P (PO4-P) by using both a "segmented-flow" analyzer and a "discrete" analyzer. Plots of the discrete vs. the segmented-flow results showed linear correlation coefficients of 0.9980 for NH3-N, 0.9997 for NO3 + NO2-N, 0.9998 for NO2-N, and 0.9950 for PO4-P. The significances of the slope and the y intercept of each plot are discussed in terms of possible biases which may exist between the two systems. Data concerning precision and accuracy for both analyzers are presented. For the four analytes, the rate of sample analysis was 60 samples per hour on the discrete analyzer and 40 samples per hour on the segmented-flow analyzer. ?? 1981 American Chemical Society.

  5. Closed formulae to determine the angular velocity of a body-segment based on 3D measurements.

    PubMed

    Kocsis, L; Béda, G

    2001-01-01

    This paper suggests a simple method to determine the global coordinates of the angular velocity and the angular acceleration of a body segment determined by the coordinates of minimum three markers. There are commonly used calculations for the angular quantities basing on the "hypothesis" of planar motion. The usage of approximate methods can result in quantitative and qualitative errors that may completely disort the reality. The method mentioned here is theoretically absolutely correct and can be well used for smoothing noisy data. PMID:11811842

  6. Drosophila spermiogenesis

    PubMed Central

    Fabian, Lacramioara; Brill, Julie A.

    2012-01-01

    Drosophila melanogaster spermatids undergo dramatic morphological changes as they differentiate from small round cells approximately 12 μm in diameter into highly polarized, 1.8 mm long, motile sperm capable of participating in fertilization. During spermiogenesis, syncytial cysts of 64 haploid spermatids undergo synchronous differentiation. Numerous changes occur at a subcellular level, including remodeling of existing organelles (mitochondria, nuclei), formation of new organelles (flagellar axonemes, acrosomes), polarization of elongating cysts and plasma membrane addition. At the end of spermatid morphogenesis, organelles, mitochondrial DNA and cytoplasmic components not needed in mature sperm are stripped away in a caspase-dependent process called individualization that results in formation of individual sperm. Here, we review the stages of Drosophila spermiogenesis and examine our current understanding of the cellular and molecular mechanisms involved in shaping male germ cell-specific organelles and forming mature, fertile sperm. PMID:23087837

  7. Segmentation-based determination of terrain points from full-waveform airborne laser scanning data

    NASA Astrophysics Data System (ADS)

    Mücke, Werner; Hollaus, Markus; Briese, Christian

    2010-05-01

    model (DTM) generation. While the amplitude, as a measure of the backscattered intensity, holds information about the reflectivity of the surface, the echo width is influenced by height variations of small scatterers within the footprint area of the laser beam. In terms of ALS, overgrown areas were found to show lower amplitudes than open terrain, due to loss of backscattered energy because of the penetration of the foliage. On the other hand, vegetation is considered to feature large variations in height within the footprint area and therefore feature larger echo widths. Hence, these two observables have potential to be used within the process of DTM derivation, eliminating echoes from vegetated areas and produce a more reliable terrain point cloud. A seeded region growing segmentation algorithm can be employed to gather points with similar FWF attributes. Further segmentation constraints can be the application of maximum height intervals, to prevent probable terrain segments from growing into vegetation. By signature analysis from training areas, thresholds for the extraction of the segments representing terrain can be provided. In addition, we assume that an acceleration in computation with conventional filtering algorithms can be achieved due to the reduced set of input points.

  8. Nonlinear physical segmentation algorithm for determining the layer boundary from lidar signal.

    PubMed

    Mao, Feiyue; Li, Jun; Li, Chen; Gong, Wei; Min, Qilong; Wang, Wei

    2015-11-30

    Layer boundary (base and top) detection is a basic problem in lidar data processing, the results of which are used as inputs of optical properties retrieval. However, traditional algorithms not only require manual intervention but also rely heavily on the signal-to-noise ratio. Therefore, we propose a robust and automatic algorithm for layer detection based on a novel algorithm for lidar signal segmentation and representation. Our algorithm is based on the lidar equation and avoids most of the limitations of the traditional algorithms. Testing of the simulated and real signals shows that the algorithm is able to position the base and top accurately even with a low signal to noise ratio. Furthermore, the results of the classification are accurate and satisfactory. The experimental results confirm that our algorithm can be used for automatic detection, retrieval, and analysis of lidar data sets. PMID:26698806

  9. Drosophila myogenesis.

    PubMed

    Bothe, Ingo; Baylies, Mary K

    2016-09-12

    The skeletal muscle system is the largest organ in motile animals, constituting between 35 and 55% of the human body mass, and up to 75% of the body mass in flying organisms like Drosophila. The flight muscles alone in flying insects comprise up to 65% of total body mass. Not only is the musculature the largest organ system, it is also exquisitely complex, with single muscles existing in different shapes and sizes. These different morphologies allow for such different functions as the high-frequency beating of a wing in a hummingbird, the dilation of the pupil in a human eye, or the maintenance of posture in a giraffe's neck. PMID:27623256

  10. Stereophotogrammetrie Mass Distribution Parameter Determination Of The Lower Body Segments For Use In Gait Analysis

    NASA Astrophysics Data System (ADS)

    Sheffer, Daniel B.; Schaer, Alex R.; Baumann, Juerg U.

    1989-04-01

    Inclusion of mass distribution information in biomechanical analysis of motion is a requirement for the accurate calculation of external moments and forces acting on the segmental joints during locomotion. Regression equations produced from a variety of photogrammetric, anthropometric and cadaeveric studies have been developed and espoused in literature. Because of limitations in the accuracy of predicted inertial properties based on the application of regression equation developed on one population and then applied on a different study population, the employment of a measurement technique that accurately defines the shape of each individual subject measured is desirable. This individual data acquisition method is especially needed when analyzing the gait of subjects with large differences in their extremity geo-metry from those considered "normal", or who may possess gross asymmetries in shape in their own contralateral limbs. This study presents the photogrammetric acquisition and data analysis methodology used to assess the inertial tensors of two groups of subjects, one with spastic diplegic cerebral palsy and the other considered normal.

  11. Dreams Fulfilled and Shattered: Determinants of Segmented Assimilation in the Second Generation*

    PubMed Central

    Haller, William; Portes, Alejandro; Lynch, Scott M.

    2013-01-01

    We summarize prior theories on the adaptation process of the contemporary immigrant second generation as a prelude to presenting additive and interactive models showing the impact of family variables, school contexts and academic outcomes on the process. For this purpose, we regress indicators of educational and occupational achievement in early adulthood on predictors measured three and six years earlier. The Children of Immigrants Longitudinal Study (CILS), used for the analysis, allows us to establish a clear temporal order among exogenous predictors and the two dependent variables. We also construct a Downward Assimilation Index (DAI), based on six indicators and regress it on the same set of predictors. Results confirm a pattern of segmented assimilation in the second generation, with a significant proportion of the sample experiencing downward assimilation. Predictors of the latter are the obverse of those of educational and occupational achievement. Significant interaction effects emerge between these predictors and early school contexts, defined by different class and racial compositions. Implications of these results for theory and policy are examined. PMID:24223437

  12. Dreams Fulfilled and Shattered: Determinants of Segmented Assimilation in the Second Generation.

    PubMed

    Haller, William; Portes, Alejandro; Lynch, Scott M

    2011-03-01

    We summarize prior theories on the adaptation process of the contemporary immigrant second generation as a prelude to presenting additive and interactive models showing the impact of family variables, school contexts and academic outcomes on the process. For this purpose, we regress indicators of educational and occupational achievement in early adulthood on predictors measured three and six years earlier. The Children of Immigrants Longitudinal Study (CILS), used for the analysis, allows us to establish a clear temporal order among exogenous predictors and the two dependent variables. We also construct a Downward Assimilation Index (DAI), based on six indicators and regress it on the same set of predictors. Results confirm a pattern of segmented assimilation in the second generation, with a significant proportion of the sample experiencing downward assimilation. Predictors of the latter are the obverse of those of educational and occupational achievement. Significant interaction effects emerge between these predictors and early school contexts, defined by different class and racial compositions. Implications of these results for theory and policy are examined. PMID:24223437

  13. The retinal determination gene eyes absent is regulated by the EGF receptor pathway throughout development in Drosophila.

    PubMed

    Salzer, Claire L; Elias, Yair; Kumar, Justin P

    2010-01-01

    Members of the Eyes absent (Eya) protein family play important roles in tissue specification and patterning by serving as both transcriptional activators and protein tyrosine phosphatases. These activities are often carried out in the context of complexes containing members of the Six and/or Dach families of DNA binding proteins. eyes absent, the founding member of the Eya family is expressed dynamically within several embryonic, larval, and adult tissues of the fruit fly, Drosophila melanogaster. Loss-of-function mutations are known to result in disruptions of the embryonic head and central nervous system as well as the adult brain and visual system, including the compound eyes. In an effort to understand how eya is regulated during development, we have carried out a genetic screen designed to identify genes that lie upstream of eya and govern its expression. We have identified a large number of putative regulators, including members of several signaling pathways. Of particular interest is the identification of both yan/anterior open and pointed, two members of the EGF Receptor (EGFR) signaling cascade. The EGFR pathway is known to regulate the activity of Eya through phosphorylation via MAPK. Our findings suggest that this pathway is also used to influence eya transcriptional levels. Together these mechanisms provide a route for greater precision in regulating a factor that is critical for the formation of a wide range of diverse tissues. PMID:19884307

  14. The Retinal Determination Gene eyes absent Is Regulated by the EGF Receptor Pathway Throughout Development in Drosophila

    PubMed Central

    Salzer, Claire L.; Elias, Yair; Kumar, Justin P.

    2010-01-01

    Members of the Eyes absent (Eya) protein family play important roles in tissue specification and patterning by serving as both transcriptional activators and protein tyrosine phosphatases. These activities are often carried out in the context of complexes containing members of the Six and/or Dach families of DNA binding proteins. eyes absent, the founding member of the Eya family is expressed dynamically within several embryonic, larval, and adult tissues of the fruit fly, Drosophila melanogaster. Loss-of-function mutations are known to result in disruptions of the embryonic head and central nervous system as well as the adult brain and visual system, including the compound eyes. In an effort to understand how eya is regulated during development, we have carried out a genetic screen designed to identify genes that lie upstream of eya and govern its expression. We have identified a large number of putative regulators, including members of several signaling pathways. Of particular interest is the identification of both yan/anterior open and pointed, two members of the EGF Receptor (EGFR) signaling cascade. The EGFR pathway is known to regulate the activity of Eya through phosphorylation via MAPK. Our findings suggest that this pathway is also used to influence eya transcriptional levels. Together these mechanisms provide a route for greater precision in regulating a factor that is critical for the formation of a wide range of diverse tissues. PMID:19884307

  15. Copy Number and Orientation Determine the Susceptibility of a Gene to Silencing by Nearby Heterochromatin in Drosophila

    PubMed Central

    Sabl, J. F.; Henikoff, S.

    1996-01-01

    The classical phenomenon of position-effect variegation (PEV) is the mosaic expression that occurs when a chromosomal rearrangement moves a euchromatic gene near heterochromatin. A striking feature of this phenomenon is that genes far away from the junction with heterochromatin can be affected, as if the heterochromatic state ``spreads.'' We have investigated classical PEV of a Drosophila brown transgene affected by a heterochromatic junction ~60 kb away. PEV was enhanced when the transgene was locally duplicated using P transposase. Successive rounds of P transposase mutagenesis and phenotypic selection produced a series of PEV alleles with differences in phenotype that depended on transgene copy number and orientation. As for other examples of classical PEV, nearby heterochromatin was required for gene silencing. Modifications of classical PEV by alterations at a single site are unexpected, and these observations contradict models for spreading that invoke propagation of heterochromatin along the chromosome. Rather, our results support a model in which local alterations affect the affinity of a gene region for nearby heterochromatin via homology-based pairing, suggesting an alternative explanation for this 65-year-old phenomenon. PMID:8852844

  16. Forces driven by morphogenesis modulate Twist Expression to determine Anterior Mid-gut Differentiation in Drosophila embryos

    NASA Astrophysics Data System (ADS)

    Farge, Emmanuel

    2008-03-01

    By combining magnetic tweezers to in vivo laser ablation, we locally manipulate Drosophila embryonic tissues with physiologically relevant forces. We demonstrate that high level of Twist expression in the stomodeal primordium is mechanically induced in response to compression by the 60±20 nN force developed during germ-band extension (GBE). We find that this force triggers the junctional release and nuclear translocation of Armadillo involved in Twist mechanical induction in the stomodeum in a Src42A dependent way. Finally, stomodeal-specific RNAi-mediated silencing of Twist during compression impairs the differentiation of midgut cells, as revealed by strong defects in Dve expression and abnormal larval lethality. Thus, mechanical induction of Twist overexpression in stomodeal cells is necessary for subsequent midgut differentiation. In collaboration with Nicolas Desprat, Willy Supatto, and Philippe-Alexandre Pouille, MGDET, UMR168 CNRS, Institut Curie11 rue Pierre et Marie Curie, F-75005, Paris, France; and Emmanuel Beaurepaire, LOB, Ecole Polytechnique, CNRS and INSERM U 696, 91128 Palaiseau, France.

  17. Application of self-organising maps towards segmentation of soybean samples by determination of amino acids concentration.

    PubMed

    Silva, Lívia Ramazzoti Chanan; Angilelli, Karina Gomes; Cremasco, Hágata; Romagnoli, Érica Signori; Galão, Olívio Fernandes; Borsato, Dionisio; Moraes, Larissa Alexandra Cardoso; Mandarino, José Marcos Gontijo

    2016-09-01

    Soybeans are widely used both for human nutrition and animal feed, since they are an important source of protein, and they also provide components such as phytosterols, isoflavones, and amino acids. In this study, were determined the concentrations of the amino acids lysine, histidine, arginine, asparagine, glutamic acid, glycine, alanine, valine, isoleucine, leucine, tyrosine, phenylalanine present in 14 samples of conventional soybeans and 6 transgenic, cultivated in two cities of the state of Paraná, Londrina and Ponta Grossa. The results were tabulated and presented to a self-organising map for segmentation according planting regions and conventional or transgenic varieties. A network with 7000 training epochs and a 10 × 10 topology was used, and it proved appropriate in the segmentation of the samples using the data analysed. The weight maps provided by the network, showed that all the amino acids were important in targeting the samples, especially isoleucine. Three clusters were formed, one with only Ponta Grossa samples (including transgenic (PGT) and common (PGC)), a second group with Londrina transgenic (LT) samples and the third with Londrina common (LC) samples. PMID:27213953

  18. On the genetic parameter determining the efficiency of purging: an estimate for Drosophila egg-to-pupae viability.

    PubMed

    Bersabé, D; García-Dorado, A

    2013-02-01

    The consequences of inbreeding on fitness can be crucial in evolutionary and conservation grounds and depend upon the efficiency of purging against deleterious recessive alleles. Recently, analytical expressions have been derived to predict the evolution of mean fitness, taking into account both inbreeding and purging, which depend on an 'effective purging coefficient (d(e) )'. Here, we explore the validity of that predictive approach and assay the strength of purging by estimating d(e) for egg-to-pupae viability (EPV) after a drastic reduction in population size in a recently captured base population of Drosophila melanogaster. For this purpose, we first obtained estimates of the inbreeding depression rate (δ) for EPV in the base population, and we found that about 40% was due to segregating recessive lethals. Then, two sets of lines were founded from this base population and were maintained with different effective size throughout the rest of the experiment (N = 6; N = 12), their mean EPV being assayed at different generations. Due to purging, the reductions in mean EPV experienced by these lines were considerably smaller than the corresponding neutral predictions. For the 60% of δ attributable to nonlethal deleterious alleles, our results suggest an effective purging coefficient d(e) > 0.02. Similarly, we obtain that d(e) > 0.09 is required to roughly account for purging against the pooled inbreeding depression from lethal and nonlethal deleterious alleles. This implies that purging should be efficient for population sizes of the order of a few tens and larger, but might be inefficient against nonlethal deleterious alleles in smaller populations. PMID:23199278

  19. A balance of positive and negative regulators determines the pace of the segmentation clock

    PubMed Central

    Wiedermann, Guy; Bone, Robert Alexander; Silva, Joana Clara; Bjorklund, Mia

    2015-01-01

    Somitogenesis is regulated by a molecular oscillator that drives dynamic gene expression within the pre-somitic mesoderm. Previous mathematical models of the somitogenesis clock that invoke the mechanism of delayed negative feedback predict that its oscillation period depends on the sum of delays inherent to negative-feedback loops and inhibitor half-lives. We develop a mathematical model that explores the possibility that positive feedback also plays a role in determining the period of clock oscillations. The model predicts that increasing the half-life of the positive regulator, Notch intracellular domain (NICD), can lead to elevated NICD levels and an increase in the oscillation period. To test this hypothesis, we investigate a phenotype induced by various small molecule inhibitors in which the clock is slowed. We observe elevated levels and a prolonged half-life of NICD. Reducing NICD production rescues these effects. These data provide the first indication that tight control of the turnover of positive as well as negative regulators of the clock determines its periodicity. DOI: http://dx.doi.org/10.7554/eLife.05842.001 PMID:26357015

  20. Complete mitochondrial genome of Drosophila albomicans.

    PubMed

    Kang, Xiongbin; Luo, Xiao; Zhang, Zhi; Zhang, Zhen; Yang, Junqing; Bi, Guiqi

    2016-09-01

    Drosophila albomicans has been widely used as an important animal model for chromosome evolution. In this study, the mitochondrial genome sequence of this species is determined and described for the first time. The mitochondrial genome (15 849 bp) encompasses two rRNA, 22 tRNA, and 13 protein-coding genes. Genome content and structure are similar to those reported from other Drosophila mitochondrial genomes. Phylogeny analysis indicates that D. albomicans have a closer genetic relationship with Drosophil aincompta and Drosophil alittoralis. This mitochondrial genome is potentially important for studying molecular evolution and conservation genetics in Drosophila genus. PMID:26358579

  1. Attractiveness of fermentation and related products to spotted wing Drosophila (Diptera: Drosophilidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Laboratory screening bioassays and field trapping experiments of spotted wing Drosophila flies, Drosophila suzukii were conducted to determine the attractiveness of 17 potentially attractive compounds as well as compare attractant efficiency during peak fruit ripeness and postharvest captures late ...

  2. Imaging fictive locomotor patterns in larval Drosophila

    PubMed Central

    Bayley, Timothy G.; Taylor, Adam L.; Berni, Jimena; Bate, Michael; Hedwig, Berthold

    2015-01-01

    We have established a preparation in larval Drosophila to monitor fictive locomotion simultaneously across abdominal and thoracic segments of the isolated CNS with genetically encoded Ca2+ indicators. The Ca2+ signals closely followed spiking activity measured electrophysiologically in nerve roots. Three motor patterns are analyzed. Two comprise waves of Ca2+ signals that progress along the longitudinal body axis in a posterior-to-anterior or anterior-to-posterior direction. These waves had statistically indistinguishable intersegmental phase delays compared with segmental contractions during forward and backward crawling behavior, despite being ∼10 times slower. During these waves, motor neurons of the dorsal longitudinal and transverse muscles were active in the same order as the muscle groups are recruited during crawling behavior. A third fictive motor pattern exhibits a left-right asymmetry across segments and bears similarities with turning behavior in intact larvae, occurring equally frequently and involving asymmetry in the same segments. Ablation of the segments in which forward and backward waves of Ca2+ signals were normally initiated did not eliminate production of Ca2+ waves. When the brain and subesophageal ganglion (SOG) were removed, the remaining ganglia retained the ability to produce both forward and backward waves of motor activity, although the speed and frequency of waves changed. Bilateral asymmetry of activity was reduced when the brain was removed and abolished when the SOG was removed. This work paves the way to studying the neural and genetic underpinnings of segmentally coordinated motor pattern generation in Drosophila with imaging techniques. PMID:26311188

  3. Imaging fictive locomotor patterns in larval Drosophila.

    PubMed

    Pulver, Stefan R; Bayley, Timothy G; Taylor, Adam L; Berni, Jimena; Bate, Michael; Hedwig, Berthold

    2015-11-01

    We have established a preparation in larval Drosophila to monitor fictive locomotion simultaneously across abdominal and thoracic segments of the isolated CNS with genetically encoded Ca(2+) indicators. The Ca(2+) signals closely followed spiking activity measured electrophysiologically in nerve roots. Three motor patterns are analyzed. Two comprise waves of Ca(2+) signals that progress along the longitudinal body axis in a posterior-to-anterior or anterior-to-posterior direction. These waves had statistically indistinguishable intersegmental phase delays compared with segmental contractions during forward and backward crawling behavior, despite being ∼10 times slower. During these waves, motor neurons of the dorsal longitudinal and transverse muscles were active in the same order as the muscle groups are recruited during crawling behavior. A third fictive motor pattern exhibits a left-right asymmetry across segments and bears similarities with turning behavior in intact larvae, occurring equally frequently and involving asymmetry in the same segments. Ablation of the segments in which forward and backward waves of Ca(2+) signals were normally initiated did not eliminate production of Ca(2+) waves. When the brain and subesophageal ganglion (SOG) were removed, the remaining ganglia retained the ability to produce both forward and backward waves of motor activity, although the speed and frequency of waves changed. Bilateral asymmetry of activity was reduced when the brain was removed and abolished when the SOG was removed. This work paves the way to studying the neural and genetic underpinnings of segmentally coordinated motor pattern generation in Drosophila with imaging techniques. PMID:26311188

  4. An Automatic Assessment System of Diabetic Foot Ulcers Based on Wound Area Determination, Color Segmentation, and Healing Score Evaluation

    PubMed Central

    Wang, Lei; Pedersen, Peder C.; Strong, Diane M.; Tulu, Bengisu; Agu, Emmanuel; Ignotz, Ron; He, Qian

    2015-01-01

    Background: For individuals with type 2 diabetes, foot ulcers represent a significant health issue. The aim of this study is to design and evaluate a wound assessment system to help wound clinics assess patients with foot ulcers in a way that complements their current visual examination and manual measurements of their foot ulcers. Methods: The physical components of the system consist of an image capture box, a smartphone for wound image capture and a laptop for analyzing the wound image. The wound image assessment algorithms calculate the overall wound area, color segmented wound areas, and a healing score, to provide a quantitative assessment of the wound healing status both for a single wound image and comparisons of subsequent images to an initial wound image. Results: The system was evaluated by assessing foot ulcers for 12 patients in the Wound Clinic at University of Massachusetts Medical School. As performance measures, the Matthews correlation coefficient (MCC) value for the wound area determination algorithm tested on 32 foot ulcer images was .68. The clinical validity of our healing score algorithm relative to the experienced clinicians was measured by Krippendorff’s alpha coefficient (KAC) and ranged from .42 to .81. Conclusion: Our system provides a promising real-time method for wound assessment based on image analysis. Clinical comparisons indicate that the optimized mean-shift-based algorithm is well suited for wound area determination. Clinical evaluation of our healing score algorithm shows its potential to provide clinicians with a quantitative method for evaluating wound healing status. PMID:26253144

  5. A theoretical model for the regulation of Sex-lethal, a gene that controls sex determination and dosage compensation in Drosophila melanogaster.

    PubMed Central

    Louis, Matthieu; Holm, Liisa; Sánchez, Lucas; Kaufman, Marcelle

    2003-01-01

    Cell fate commitment relies upon making a choice between different developmental pathways and subsequently remembering that choice. Experimental studies have thoroughly investigated this central theme in biology for sex determination. In the somatic cells of Drosophila melanogaster, Sex-lethal (Sxl) is the master regulatory gene that specifies sexual identity. We have developed a theoretical model for the initial sex-specific regulation of Sxl expression. The model is based on the well-documented molecular details of the system and uses a stochastic formulation of transcription. Numerical simulations allow quantitative assessment of the role of different regulatory mechanisms in achieving a robust switch. We establish on a formal basis that the autoregulatory loop involved in the alternative splicing of Sxl primary transcripts generates an all-or-none bistable behavior and constitutes an efficient stabilization and memorization device. The model indicates that production of a small amount of early Sxl proteins leaves the autoregulatory loop in its off state. Numerical simulations of mutant genotypes enable us to reproduce and explain the phenotypic effects of perturbations induced in the dosage of genes whose products participate in the early Sxl promoter activation. PMID:14668388

  6. Automated Tracking of Drosophila Specimens

    PubMed Central

    Chao, Rubén; Macía-Vázquez, Germán; Zalama, Eduardo; Gómez-García-Bermejo, Jaime; Perán, José-Ramón

    2015-01-01

    The fruit fly Drosophila Melanogaster has become a model organism in the study of neurobiology and behavior patterns. The analysis of the way the fly moves and its behavior is of great scientific interest for research on aspects such as drug tolerance, aggression or ageing in humans. In this article, a procedure for detecting, identifying and tracking numerous specimens of Drosophila by means of computer vision-based sensing systems is presented. This procedure allows dynamic information about each specimen to be collected at each moment, and then for its behavior to be quantitatively characterized. The proposed algorithm operates in three main steps: a pre-processing step, a detection and segmentation step, and tracking shape. The pre-processing and segmentation steps allow some limits of the image acquisition system and some visual artifacts (such as shadows and reflections) to be dealt with. The improvements introduced in the tracking step allow the problems corresponding to identity loss and swaps, caused by the interaction between individual flies, to be solved efficiently. Thus, a robust method that compares favorably to other existing methods is obtained. PMID:26258779

  7. Automated Tracking of Drosophila Specimens.

    PubMed

    Chao, Rubén; Macía-Vázquez, Germán; Zalama, Eduardo; Gómez-García-Bermejo, Jaime; Perán, José-Ramón

    2015-01-01

    The fruit fly Drosophila Melanogaster has become a model organism in the study of neurobiology and behavior patterns. The analysis of the way the fly moves and its behavior is of great scientific interest for research on aspects such as drug tolerance, aggression or ageing in humans. In this article, a procedure for detecting, identifying and tracking numerous specimens of Drosophila by means of computer vision-based sensing systems is presented. This procedure allows dynamic information about each specimen to be collected at each moment, and then for its behavior to be quantitatively characterized. The proposed algorithm operates in three main steps: a pre-processing step, a detection and segmentation step, and tracking shape. The pre-processing and segmentation steps allow some limits of the image acquisition system and some visual artifacts (such as shadows and reflections) to be dealt with. The improvements introduced in the tracking step allow the problems corresponding to identity loss and swaps, caused by the interaction between individual flies, to be solved efficiently. Thus, a robust method that compares favorably to other existing methods is obtained. PMID:26258779

  8. Methods to assay Drosophila behavior.

    PubMed

    Nichols, Charles D; Becnel, Jaime; Pandey, Udai B

    2012-01-01

    Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases(1). We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials(2-4). The rapid iterative negative geotaxis (RING) assay(5) has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously

  9. Rhodopsin Gene Expression Determines Rod Outer Segment Size and Rod Cell Resistance to a Dominant-Negative Neurodegeneration Mutant

    PubMed Central

    Price, Brandee A.; Sandoval, Ivette M.; Chan, Fung; Nichols, Ralph; Roman-Sanchez, Ramon; Wensel, Theodore G.; Wilson, John H.

    2012-01-01

    Two outstanding unknowns in the biology of photoreceptors are the molecular determinants of cell size, which is remarkably uniform among mammalian species, and the mechanisms of rod cell death associated with inherited neurodegenerative blinding diseases such as retinitis pigmentosa. We have addressed both questions by performing an in vivo titration with rhodopsin gene copies in genetically engineered mice that express only normal rhodopsin or an autosomal dominant allele, encoding rhodopsin with a disease-causing P23H substitution. The results reveal that the volume of the rod outer segment is proportional to rhodopsin gene expression; that P23H-rhodopsin, the most common rhodopsin gene disease allele, causes cell death via a dominant-negative mechanism; and that long term survival of rod cells carrying P23H-rhodopsin can be achieved by increasing the levels of wild type rhodopsin. These results point to promising directions in gene therapy for autosomal dominant neurodegenerative diseases caused by dominant-negative mutations. PMID:23185477

  10. Temporal and spatial distribution of transcripts from the Deformed gene of Drosophila

    PubMed Central

    Chadwick, Robin; McGinnis, William

    1987-01-01

    The Deformed gene of Drosophila is necessary for the proper development of epidermal pattern elements arising from the maxillary and mandibular segments of the head. We find one major transcript (2.8 kb) homologous to Deformed (Dfd) probes which is expressed continuously from 3 h of embryogenesis into adulthood. Localized transcript accumulation is first detected just prior to the formation of the cellular blastoderm in a single circumferential band at about 65-75% egg length. The zone of Dfd expression is approximately two segment primordia in width. At later stages of embryogenesis, Dfd transcripts accumulate in the posterior ectoderm of the mandibular segment, and in the ventro-lateral ectoderm of the maxillary segment. Transcripts are also detected in the mesoderm and neuromeres of the mandibular and maxillary segments. The distribution of Dfd transcripts supports the hypothesis that Dfd functions as a homeotic selector gene in the determination of posterior head segments. ImagesFig. 1.Fig. 2.Fig. 3.Fig. 4.Fig. 5.Fig. 6.Fig. 7. PMID:16453753

  11. A large family of putative transmembrane receptors homologous to the product of the Drosophila tissue polarity gene frizzled.

    PubMed

    Wang, Y; Macke, J P; Abella, B S; Andreasson, K; Worley, P; Gilbert, D J; Copeland, N G; Jenkins, N A; Nathans, J

    1996-02-23

    In Drosophila melanogaster, the frizzled gene plays an essential role in the development of tissue polarity as assessed by the orientation of cuticular structures. Through a combination of random cDNA sequencing, degenerate polymerase chain reaction amplification, and low stringency hybridization we have identified six novel frizzled homologues from mammals, at least 11 from zebrafish, several from chicken and sea urchin, and one from Caenorhabditis elegans. The complete deduced amino acid sequences of the mammalian and nematode homologues share with the Drosophila frizzled protein a conserved amino-terminal cysteine-rich domain and seven putative transmembrane segments. Each of the mammalian homologues is expressed in a distinctive set of tissues in the adult, and at least three are expressed during embryogenesis. As hypothesized for the Drosophila frizzled protein, the frizzled homologues are likely to act as transmembrane receptors for as yet unidentified ligands. These observations predict the existence of a family of signal transduction pathways that are homologous to the pathway that determines tissue polarity in Drosophila. PMID:8626800

  12. Influence of filter choice on 18F-FDG PET segmentation accuracy determined using generalized estimating equations.

    PubMed

    McGurk, Ross J; Smith, Valerie A; Bowsher, James; Lee, John A; Das, Shiva K

    2013-06-01

    This study aims to quantify how filter choice affects several fluoro-deoxy-glucose (FDG)-positron emission tomography (PET) segmentation methods and present the use of model fitting via generalized estimating equations (GEEs) to appropriately account for the properties of a common segmentation quality metric (Dice similarity coefficient). Spherical and irregularly shaped 'hot' objects filled with 18F-FDG were placed in a medium with background activity and imaged for 1, 2 and 5 min durations at low and high contrasts. Images were filtered with Gaussian and bilateral filters of 5 and 7 mm full-width half maximum (FWHM), with and without 3 mm FWHM Gaussian pre-smoothing. Four segmentation methods were used: 40% thresholding, adaptive thresholding, k-means clustering and seeded region-growing. Segmentation accuracy was quantified by overlap (using Dice similarity coefficient (DSC)) and distance between surfaces (using symmetric-mean-absolute-surface-distance (SMASD)) of the ground truth and segmented volumes. All segmentation methods showed mean DSC values between 0.71-0.87 and mean SMASD values between 0.72-2.10 mm across filters. The bilateral filter with 3 mm FWHM Gaussian pre-smoothing had mean DSC 0.80 ± 0.17 and mean SMASD 1.17 ± 1.51 mm displaying approximately equal performance to a 5 mm Gaussian filter with mean DSC 0.79 ± 0.18 and mean SMASD 1.27 ± 1.52 mm. Results from models fit using GEE with a binomial distribution and exchangeable correlation structure estimated the correlation between DSC values as 0.118 and 0.290 for spheres and irregular objects, respectively. The GEE approach accounts for several factors specific to the DSC metric that simpler statistical approaches do not, providing more accurate estimations of experimental effects commonly associated with nuclear medicine segmentation studies. PMID:23632261

  13. Influence of filter choice on 18F-FDG PET segmentation accuracy determined using generalized estimating equations

    NASA Astrophysics Data System (ADS)

    McGurk, Ross J.; Smith, Valerie A.; Bowsher, James; Lee, John A.; Das, Shiva K.

    2013-06-01

    This study aims to quantify how filter choice affects several fluoro-deoxy-glucose (FDG)-positron emission tomography (PET) segmentation methods and present the use of model fitting via generalized estimating equations (GEEs) to appropriately account for the properties of a common segmentation quality metric (Dice similarity coefficient). Spherical and irregularly shaped ‘hot’ objects filled with 18F-FDG were placed in a medium with background activity and imaged for 1, 2 and 5 min durations at low and high contrasts. Images were filtered with Gaussian and bilateral filters of 5 and 7 mm full-width half maximum (FWHM), with and without 3 mm FWHM Gaussian pre-smoothing. Four segmentation methods were used: 40% thresholding, adaptive thresholding, k-means clustering and seeded region-growing. Segmentation accuracy was quantified by overlap (using Dice similarity coefficient (DSC)) and distance between surfaces (using symmetric-mean-absolute-surface-distance (SMASD)) of the ground truth and segmented volumes. All segmentation methods showed mean DSC values between 0.71-0.87 and mean SMASD values between 0.72-2.10 mm across filters. The bilateral filter with 3 mm FWHM Gaussian pre-smoothing had mean DSC 0.80 ± 0.17 and mean SMASD 1.17 ± 1.51 mm displaying approximately equal performance to a 5 mm Gaussian filter with mean DSC 0.79 ± 0.18 and mean SMASD 1.27 ± 1.52 mm. Results from models fit using GEE with a binomial distribution and exchangeable correlation structure estimated the correlation between DSC values as 0.118 and 0.290 for spheres and irregular objects, respectively. The GEE approach accounts for several factors specific to the DSC metric that simpler statistical approaches do not, providing more accurate estimations of experimental effects commonly associated with nuclear medicine segmentation studies.

  14. Vital Genes That Flank Sex-Lethal, an X-Linked Sex-Determining Gene of DROSOPHILA MELANOGASTER

    PubMed Central

    Nicklas, Janice A.; Cline, Thomas W.

    1983-01-01

    The X-chromosome:autosome balance in D. melanogaster appears to control both sex determination and dosage compensation through effects on a maternally influenced sex-linked gene called Sex-lethal (Sxl; 1-19.2). To facilitate molecular and genetic analysis of Sxl, we attempted to determine the locations of all ethyl methanesulfonate (EMS)-mutable genes vital to both sexes in the region between 6E1 and 7B1. This area includes approximately 1 cM of the genetic map on each side of Sxl and was reported by C. B. Bridges to contain 26 salivary gland polytene chromosome bands. The region appears rather sparsely populated with genes vital to both sexes, since the 122 recessive lethal mutations we recovered fell into only nine complementation groups. From one to 38 alleles of each gene were recovered. There was a preponderance of embryonic lethals in this area, although the lethal periods of loss-of-function mutations included larval, pupal and adult stages as well. Since the screen required that mutations be recessive and lethal to males, our failure to recover new Sxl alleles was the result expected for a gene with a female-specific function. An attempt was made to identify recessive male-specific lethals in this region, but none were found. Precise map positions were determined for eight of the nine vital genes. An interesting feature of the map is the location of Sxl in the middle of a 0.6- to 0.7-cM interval that appears to be devoid of genes vital to both sexes. The genetic location was determined of breakpoints near Sxl for all available chromosome rearrangements. Sxl is most likely located just to the left of band 7A1. We determined the relationship of our EMS-induced mutations in these nine genes to alleles induced by others. From this we conclude that the various genes appear to differ significantly from each other in their relative sensitivity to mutation by EMS vs. X rays. PMID:17246118

  15. Stand Still, a Drosophila Gene Involved in the Female Germline for Proper Survival, Sex Determination and Differentiation

    PubMed Central

    Pennetta, G.; Pauli, D.

    1997-01-01

    We identified a new gene, stand still (stil), required in the female germline for proper survival, sex determination and differentiation. Three strong loss-of-function alleles were isolated. The strongest phenotype exhibited by ovaries dissected from adult females is the complete absence of germ cells. In other ovaries, the few surviving germ cells frequently show a morphology typical of primary spermatocytes. still is not required either for fly viability or for male germline development. The gene was cloned and found to encode a novel protein. still is strongly expressed in the female germ cells. Using P[stil(+)] transgenes, we show that stil and a closely localized gene are involved in the modification of the ovarian phenotypes of the dominant alleles of ovo caused by heterozygosity of region 49 A-D. The similarity of the mutant phenotypes of stil to that of otu and ovo suggests that the three genes function in a common or in parallel pathways necessary in the female germline for its survival, sex determination and differentiation. PMID:9093851

  16. PhyloFlu, a DNA Microarray for Determining the Phylogenetic Origin of Influenza A Virus Gene Segments and the Genomic Fingerprint of Viral Strains

    PubMed Central

    Paulin, Luis F.; Soto-Del Río, María de los D.; Sánchez, Iván; Hernández, Jesús; Gutiérrez-Ríos, Rosa M.; López-Martínez, Irma; Wong-Chew, Rosa M.; Parissi-Crivelli, Aurora; Isa, P.; López, Susana

    2014-01-01

    Recent evidence suggests that most influenza A virus gene segments can contribute to the pathogenicity of the virus. In this regard, the hemagglutinin (HA) subtype of the circulating strains has been closely surveyed, but the reassortment of internal gene segments is usually not monitored as a potential source of an increased pathogenicity. In this work, an oligonucleotide DNA microarray (PhyloFlu) designed to determine the phylogenetic origins of the eight segments of the influenza virus genome was constructed and validated. Clades were defined for each segment and also for the 16 HA and 9 neuraminidase (NA) subtypes. Viral genetic material was amplified by reverse transcription-PCR (RT-PCR) with primers specific to the conserved 5′ and 3′ ends of the influenza A virus genes, followed by PCR amplification with random primers and Cy3 labeling. The microarray unambiguously determined the clades for all eight influenza virus genes in 74% (28/38) of the samples. The microarray was validated with reference strains from different animal origins, as well as from human, swine, and avian viruses from field or clinical samples. In most cases, the phylogenetic clade of each segment defined its animal host of origin. The genomic fingerprint deduced by the combined information of the individual clades allowed for the determination of the time and place that strains with the same genomic pattern were previously reported. PhyloFlu is useful for characterizing and surveying the genetic diversity and variation of animal viruses circulating in different environmental niches and for obtaining a more detailed surveillance and follow up of reassortant events that can potentially modify virus pathogenicity. PMID:24353006

  17. Determination of the 30-year creep trend on the Ismetpaşa segment of the North Anatolian Fault using an old geodetic network

    NASA Astrophysics Data System (ADS)

    Kutoglu, H. S.; Akcin, H.

    2006-08-01

    The Ismetpaşa segment of the North Anatolian Fault was ruptured during both the 1944 (Mw = 7.2) Gerede and 1951 (Mw = 6.9) Kuršunlu earthquakes. The field studies carried out in the aftermath of these two major earthquakes showed that the Ismetpaša segment had experienced a creep movement. To monitor the surface creep, a geodetic network with six control points was established on the segment. This network was observed three times - in 1972, 1982 and 1992. Based on our evaluations of those observations, the creep on the segment was geodetically determined to be 1.02 cm/year (1972-1982) and 0.93 cm/year (1982-1992) respectively. In 1999, the North Anatolian Fault experienced two major shocks - the Mw = 7.4 Gölcük and Mw = 7.2 Düzce earthquakes - both on the western part of the Ismetpaša fault. Using the global positioning system, our surveying team observed the network one more time in 2002 to assess whether these earthquakes affected the creep of the Ismetpaša segment, or not. The evaluation of the observations revealed a creep of 0.78 cm/year for the period 1992-2002. This result reveals that the creep of the segment has decreased in a linear fashion between 1972 and 2002 and that it had not been triggered by the Gölcük and Düzce earthquakes.

  18. Evidence that Egfr contributes to cryptic genetic variation for photoreceptor determination in natural populations of Drosophila melanogaster.

    PubMed

    Dworkin, Ian; Palsson, Arnar; Birdsall, Kelli; Gibson, Greg

    2003-10-28

    One objective of quantitative genetics is to identify the nucleotide variants within genes that contribute to phenotypic variation and susceptibility [1]. In an evolutionary context, this means characterizing the molecular polymorphisms that modify the penetrance and expressivity of perturbed traits. A survey of association between 267 SNPs in almost 11 kb of the D. melanogaster Egfr and the degree of eye roughening due to a gain-of-function Egfr(E1) allele crossed into 210 isogenic wild-type lines provides evidence that a handful of synonymous substitutions supply cryptic variation for photoreceptor determination. Ten sites exceed Bonferroni threshold for association in two sets of crosses to different Egfr(E1) backgrounds including a particularly significant cluster of sites in tight linkage disequilibrium toward the 3' end of the coding region. Epistatic interaction of this cluster with one other site enhances the expressivity of this haplotype. Replication of the strongest associations with an independent sample of 302 phenotypically extreme individuals derived from 1000 crosses of Egfr(E1) to freshly trapped males was achieved using modified case-control and transmission-disequilibrium tests. A tendency for the rarer alleles to have more disrupted eye development suggests that mutation-selection balance is a possible mechanism contributing to maintaining cryptic variation for Egfr. PMID:14588245

  19. Optogenetics in Drosophila Neuroscience.

    PubMed

    Riemensperger, Thomas; Kittel, Robert J; Fiala, André

    2016-01-01

    Optogenetic techniques enable one to target specific neurons with light-sensitive proteins, e.g., ion channels, ion pumps, or enzymes, and to manipulate their physiological state through illumination. Such artificial interference with selected elements of complex neuronal circuits can help to determine causal relationships between neuronal activity and the effect on the functioning of neuronal circuits controlling animal behavior. The advantages of optogenetics can best be exploited in genetically tractable animals whose nervous systems are, on the one hand, small enough in terms of cell numbers and to a certain degree stereotypically organized, such that distinct and identifiable neurons can be targeted reproducibly. On the other hand, the neuronal circuitry and the behavioral repertoire should be complex enough to enable one to address interesting questions. The fruit fly Drosophila melanogaster is a favorable model organism in this regard. However, the application of optogenetic tools to depolarize or hyperpolarize neurons through light-induced ionic currents has been difficult in adult flies. Only recently, several variants of Channelrhodopsin-2 (ChR2) have been introduced that provide sufficient light sensitivity, expression, and stability to depolarize central brain neurons efficiently in adult Drosophila. Here, we focus on the version currently providing highest photostimulation efficiency, ChR2-XXL. We exemplify the use of this optogenetic tool by applying it to a widely used aversive olfactory learning paradigm. Optogenetic activation of a population of dopamine-releasing neurons mimics the reinforcing properties of a punitive electric shock typically used as an unconditioned stimulus. In temporal coincidence with an odor stimulus this artificially induced neuronal activity causes learning of the odor signal, thereby creating a light-induced memory. PMID:26965122

  20. Homotypic Regulatory Clusters in Drosophila

    PubMed Central

    Lifanov, Alexander P.; Makeev, Vsevolod J.; Nazina, Anna G.; Papatsenko, Dmitri A.

    2003-01-01

    Cis-regulatory modules (CRMs) are transcription regulatory DNA segments (∼1 Kb range) that control the expression of developmental genes in higher eukaryotes. We analyzed clustering of known binding motifs for transcription factors (TFs) in over 60 known CRMs from 20 Drosophila developmental genes, and we present evidence that each type of recognition motif forms significant clusters within the regulatory regions regulated by the corresponding TF. We demonstrate how a search with a single binding motif can be applied to explore gene regulatory networks and to discover coregulated genes in the genome. We also discuss the potential of the clustering method in interpreting the differential response of genes to various levels of transcriptional regulators. PMID:12670999

  1. Spatial dependence of predictions from image segmentation: a methods to determine appropriate scales for producing land-management information

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A challenge in ecological studies is defining scales of observation that correspond to relevant ecological scales for organisms or processes. Image segmentation has been proposed as an alternative to pixel-based methods for scaling remotely-sensed data into ecologically-meaningful units. However, to...

  2. Development of dendrite polarity in Drosophila neurons

    PubMed Central

    2012-01-01

    Background Drosophila neurons have dendrites that contain minus-end-out microtubules. This microtubule arrangement is different from that of cultured mammalian neurons, which have mixed polarity microtubules in dendrites. Results To determine whether Drosophila and mammalian dendrites have a common microtubule organization during development, we analyzed microtubule polarity in Drosophila dendritic arborization neuron dendrites at different stages of outgrowth from the cell body in vivo. As dendrites initially extended, they contained mixed polarity microtubules, like mammalian neurons developing in culture. Over a period of several days this mixed microtubule array gradually matured to a minus-end-out array. To determine whether features characteristic of dendrites were localized before uniform polarity was attained, we analyzed dendritic markers as dendrites developed. In all cases the markers took on their characteristic distribution while dendrites had mixed polarity. An axonal marker was also quite well excluded from dendrites throughout development, although this was perhaps more efficient in mature neurons. To confirm that dendrite character could be acquired in Drosophila while microtubules were mixed, we genetically disrupted uniform dendritic microtubule organization. Dendritic markers also localized correctly in this case. Conclusions We conclude that developing Drosophila dendrites initially have mixed microtubule polarity. Over time they mature to uniform microtubule polarity. Dendrite identity is established before the mature microtubule arrangement is attained, during the period of mixed microtubule polarity. PMID:23111238

  3. Xotch, the Xenopus homolog of Drosophila notch.

    PubMed

    Coffman, C; Harris, W; Kintner, C

    1990-09-21

    During the development of a vertebrate embryo, cell fate is determined by inductive signals passing between neighboring tissues. Such determinative interactions have been difficult to characterize fully without knowledge of the molecular mechanisms involved. Mutations of Drosophila and the nematode Caenorhabditis elegans have been isolated that define a family of related gene products involved in similar types of cellular inductions. One of these genes, the Notch gene from Drosophila, is involved with cell fate choices in the neurogenic region of the blastoderm, in the developing nervous system, and in the eye-antennal imaginal disc. Complementary DNA clones were isolated from Xenopus embryos with Notch DNA in order to investigate whether cell-cell interactions in vertebrate embryos also depend on Notch-like molecules. This approach identified a Xenopus molecule, Xotch, which is remarkably similar to Drosophila Notch in both structure and developmental expression. PMID:2402639

  4. Segmental neurofibromatosis.

    PubMed

    Galhotra, Virat; Sheikh, Soheyl; Jindal, Sanjeev; Singla, Anshu

    2014-07-01

    Segmental neurofibromatosis is a rare disorder, characterized by neurofibromas or cafι-au-lait macules limited to one region of the body. Its occurrence on the face is extremely rare and only few cases of segmental neurofibromatosis over the face have been described so far. We present a case of segmental neurofibromatosis involving the buccal mucosa, tongue, cheek, ear, and neck on the right side of the face. PMID:25565748

  5. The Drosophila visual system

    PubMed Central

    Zhu, Yan

    2013-01-01

    A compact genome and a tiny brain make Drosophila the prime model to understand the neural substrate of behavior. The neurogenetic efforts to reveal neural circuits underlying Drosophila vision started about half a century ago, and now the field is booming with sophisticated genetic tools, rich behavioral assays, and importantly, a greater number of scientists joining from different backgrounds. This review will briefly cover the structural anatomy of the Drosophila visual system, the animal’s visual behaviors, the genes involved in assembling these circuits, the new and powerful techniques, and the challenges ahead for ultimately identifying the general principles of biological computation in the brain.   A typical brain utilizes a great many compact neural circuits to collect and process information from the internal biological and external environmental worlds and generates motor commands for observable behaviors. The fruit fly Drosophila melanogaster, despite of its miniature body and tiny brain, can survive in almost any corner of the world.1 It can find food, court mate, fight rival conspecific, avoid predators, and amazingly fly without crashing into trees. Drosophila vision and its underlying neuronal machinery has been a key research model for at least half century for neurogeneticists.2 Given the efforts invested on the visual system, this animal model is likely to offer the first full understanding of how visual information is computed by a multi-cellular organism. Furthermore, research in Drosophila has revealed many genes that play crucial roles in the formation of functional brains across species. The architectural similarities between the visual systems of Drosophila and vertebrate at the molecular, cellular, and network levels suggest new principles discovered at the circuit level on the relationship between neurons and behavior in Drosophila shall also contribute greatly to our understanding of the general principles for how bigger brains work.3

  6. Drosophila Blastorderm Analysis Software

    SciTech Connect

    2006-10-25

    PointCloudMake analyzes 3D fluorescent images of whole Drosophila embryo and produces a table-style "PointCloud" file which contains the coordinates and volumes of all the nuclei, cells, their associated relative gene expression levels along with morphological features of the embryo. See: Luengo Hendrix et at 2006 3D Morphology and Gene Expression in the Drosophila Blastoderm at Cellular Resolution manuscript submitted LBNL # LBNL-60178 Knowles DW, Keranen SVE, Biggin M. Sudar S (2002) Mapping organism expression levels at cellular resolution in developing Drosophila. In: Conchello JA, Cogswell CJ, Wilson T, editors. Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX. pp. 57-64

  7. Tension, cell shape and triple-junction angle anisotropy in the Drosophila germband

    NASA Astrophysics Data System (ADS)

    Lacy, Monica; Hutson, M. Shane; Meyer, Christian; McDonald, Xena

    In the field of tissue mechanics, the embryonic development of Drosophila melanogaster offers many opportunities for study. One of Drosophila's most crucial morphogenetic stages is the retraction of an epithelial tissue called the germband. During retraction, the segments of the retracting germband, as well as the individual germband cells, elongate in response to forces from a connected tissue, the amnioserosa. Modeling of this elongation, based on tissue responses to laser wounding, has plotted the internal germband tension against the external amnioserosa stress, creating a phase space to determine points and regions corresponding to stable elongation. Although the resulting fits indicate a necessary opposition of internal and external forces, they are inconclusive regarding the exact balance. We will present results testing the model predictions by measuring cell shapes and the correlations between cell-edge directions and triple-junction angles. These measures resolve the ambiguity in pinpointing the internal-external force balance for each germband segment. Research was supported by NIH Grant Numbers 1R01GM099107 and 1R21AR068933.

  8. Segmental neurofibromatosis.

    PubMed

    Toy, Brian

    2003-10-01

    Segmental neurofibromatosis is a rare variant of neurofibromatosis in which skin lesions are confined to a circumscribed body segment. A case of a 72-year-old woman with this condition is presented. Clinical features and genetic evidence are reviewed. PMID:14594599

  9. Active Segmentation

    PubMed Central

    Mishra, Ajay; Aloimonos, Yiannis

    2009-01-01

    The human visual system observes and understands a scene/image by making a series of fixations. Every fixation point lies inside a particular region of arbitrary shape and size in the scene which can either be an object or just a part of it. We define as a basic segmentation problem the task of segmenting that region containing the fixation point. Segmenting the region containing the fixation is equivalent to finding the enclosing contour- a connected set of boundary edge fragments in the edge map of the scene - around the fixation. This enclosing contour should be a depth boundary. We present here a novel algorithm that finds this bounding contour and achieves the segmentation of one object, given the fixation. The proposed segmentation framework combines monocular cues (color/intensity/texture) with stereo and/or motion, in a cue independent manner. The semantic robots of the immediate future will be able to use this algorithm to automatically find objects in any environment. The capability of automatically segmenting objects in their visual field can bring the visual processing to the next level. Our approach is different from current approaches. While existing work attempts to segment the whole scene at once into many areas, we segment only one image region, specifically the one containing the fixation point. Experiments with real imagery collected by our active robot and from the known databases 1 demonstrate the promise of the approach. PMID:20686671

  10. In focus: spotted wing drosophila, Drosophila suzukii, across perspectives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An effective response to the invasion of spotted wing Drosophila (SWD), Drosophila suzukii, requires proper taxonomic identification at the initial phase, understanding its basic biology and phenology, developing management tools, transferring information and technology quickly to user groups, and e...

  11. A connectionist model of the Drosophila blastoderm

    SciTech Connect

    Reinitz, J. . Dept. of Biological Sciences); Mjolsness, E. . Dept. of Computer Science); Sharp, D.H. . Theoretical Div.)

    1990-11-01

    The authors present a phenomenological modeling framework for development, and apply it to the network of segmentation genes operating in the blastoderm of Drosophila. Their purpose is to provide a systematic method for discovering and expressing correlations in experimental data on gene expression and other developmental processes. The modeling framework is based on a connectionist or neural net dynamics for biochemical regulators, coupled to grammatical rules which describe certain features of the birth, growth, and death of cells, synapses and other biological entities. They present preliminary numerical results regarding regulatory interactions between the genes Kruppel and knirps that demonstrate the potential utility of the model. 14 refs., 5 figs.

  12. Segmentation of stereo terrain images

    NASA Astrophysics Data System (ADS)

    George, Debra A.; Privitera, Claudio M.; Blackmon, Theodore T.; Zbinden, Eric; Stark, Lawrence W.

    2000-06-01

    We have studied four approaches to segmentation of images: three automatic ones using image processing algorithms and a fourth approach, human manual segmentation. We were motivated toward helping with an important NASA Mars rover mission task -- replacing laborious manual path planning with automatic navigation of the rover on the Mars terrain. The goal of the automatic segmentations was to identify an obstacle map on the Mars terrain to enable automatic path planning for the rover. The automatic segmentation was first explored with two different segmentation methods: one based on pixel luminance, and the other based on pixel altitude generated through stereo image processing. The third automatic segmentation was achieved by combining these two types of image segmentation. Human manual segmentation of Martian terrain images was used for evaluating the effectiveness of the combined automatic segmentation as well as for determining how different humans segment the same images. Comparisons between two different segmentations, manual or automatic, were measured using a similarity metric, SAB. Based on this metric, the combined automatic segmentation did fairly well in agreeing with the manual segmentation. This was a demonstration of a positive step towards automatically creating the accurate obstacle maps necessary for automatic path planning and rover navigation.

  13. Genetic control of segmentation of axial structures in vertebrates

    SciTech Connect

    Mglinets, V.A.

    1995-07-01

    The processes of segmentation of axial structures in vertebrates during early embryonic development are reviewed. These processes include the formation of neuromeres, somitomeres, cranial ganglia, and branchial arches in the head and of neuromeres, somites, spinal ganglia, and motor nerves in the body of the embryo. The class of vertebrate homeobox genes Hox is described with respect to the arrangement of these genes in four clusters, the structural and functional similarity of paralogues in gene subfamilies, and the type of Hox gene expression in the head and body. A hypothesis concerning the existence of a genetic Hox code, determining the fate of individual segments in neuroectodermal and mesenchymal derivatives, is discussed. In the context of this hypothesis, phenotypic expression of the mutant Hox genes, accompanied by the loss of their function and cases of excessive and ectopic expression of Hox genes, are analyzed. Only in such cases do mutant phenotypes demonstrate symptoms of actual homeotic transformation, in which specific segmental structures are substituted by their homologues, as has been described for homeotic mutations in Drosophila. 56 refs., 1 fig.

  14. Flying Drosophila orient to sky polarization.

    PubMed

    Weir, Peter T; Dickinson, Michael H

    2012-01-10

    Insects maintain a constant bearing across a wide range of spatial scales. Monarch butterflies and locusts traverse continents [1, 2], and foraging bees and ants travel hundreds of meters to return to their nests [1, 3, 4], whereas many other insects fly straight for only a few centimeters before changing direction. Despite this variation in spatial scale, the brain region thought to underlie long-distance navigation is remarkably conserved [5, 6], suggesting that the use of a celestial compass is a general and perhaps ancient capability of insects. Laboratory studies of Drosophila have identified a local search mode in which short, straight segments are interspersed with rapid turns [7, 8]. However, this flight mode is inconsistent with measured gene flow between geographically separated populations [9-11], and individual Drosophila can travel 10 km across desert terrain in a single night [9, 12, 13]-a feat that would be impossible without prolonged periods of straight flight. To directly examine orientation behavior under outdoor conditions, we built a portable flight arena in which a fly viewed the natural sky through a liquid crystal device that could experimentally rotate the polarization angle. Our findings indicate that Drosophila actively orient using the sky's natural polarization pattern. PMID:22177905

  15. Identification of common excitatory motoneurons in Drosophila melanogaster larvae.

    PubMed

    Takizawa, Eiji; Komatsu, Akira; Tsujimura, Hidenobu

    2007-05-01

    In insects, four types of motoneurons have long been known, including fast motoneurons, slow motoneurons, common inhibitory motoneurons, and DUM neurons. They innervate the same muscle and control its contraction together. Recent studies in Drosophila have suggested the existence of another type of motoneuron, the common excitatory motoneuron. Here, we found that shakB-GAL4 produced by labels this type of motoneuron in Drosophila larvae. We found that Drosophila larvae have two common excitatory motoneurons in each abdominal segment, RP2 for dorsal muscles and MNSNb/d-Is for ventral muscles. They innervate most of the internal longitudinal or oblique muscles on the dorsal or ventral body wall with type-Is terminals and use glutamate as a transmitter. Electrophysiological recording indicated that stimulation of the RP2 axon evoked excitatory junctional potential in a dorsal muscle. PMID:17867850

  16. A novel, tissue-specific, Drosophila homeobox gene.

    PubMed Central

    Barad, M; Jack, T; Chadwick, R; McGinnis, W

    1988-01-01

    The homeobox gene family of Drosophila appears to control a variety of position-specific patterning decisions during embryonic and imaginal development. Most of these patterning decisions determine groups of cells on the anterior-posterior axis of the Drosophila germ band. We have isolated a novel homeobox gene from Drosophila, designated H2.0. H2.0 has the most diverged homeobox so far characterized in metazoa, and, in contrast to all previously isolated homeobox genes, H2.0 exhibits a tissue-specific pattern of expression. The cells that accumulate transcripts for this novel gene correspond to the visceral musculature and its anlagen. Images PMID:2901348

  17. The segment polarity network is a robust developmental module

    NASA Astrophysics Data System (ADS)

    von Dassow, George; Meir, Eli; Munro, Edwin M.; Odell, Garrett M.

    2000-07-01

    All insects possess homologous segments, but segment specification differs radically among insect orders. In Drosophila, maternal morphogens control the patterned activation of gap genes, which encode transcriptional regulators that shape the patterned expression of pair-rule genes. This patterning cascade takes place before cellularization. Pair-rule gene products subsequently `imprint' segment polarity genes with reiterated patterns, thus defining the primordial segments. This mechanism must be greatly modified in insect groups in which many segments emerge only after cellularization. In beetles and parasitic wasps, for instance, pair-rule homologues are expressed in patterns consistent with roles during segmentation, but these patterns emerge within cellular fields. In contrast, although in locusts pair-rule homologues may not control segmentation, some segment polarity genes and their interactions are conserved. Perhaps segmentation is modular, with each module autonomously expressing a characteristic intrinsic behaviour in response to transient stimuli. If so, evolution could rearrange inputs to modules without changing their intrinsic behaviours. Here we suggest, using computer simulations, that the Drosophila segment polarity genes constitute such a module, and that this module is resistant to variations in the kinetic constants that govern its behaviour.

  18. Molecular characterization of bsg25D: a blastoderm-specific locus of Drosophila melanogaster.

    PubMed Central

    Boyer, P D; Mahoney, P A; Lengyel, J A

    1987-01-01

    The blastoderm stage of Drosophila embryogenesis is a time of crucial transitions in RNA transcription, the cell cycle and segment determination. We have previously identified three loci encoding RNAs specific to this stage (Roark et al., Dev. Biol. 109, 476-488, 1985). We present here the complete nucleotide sequence of one of these loci, bsg25D, which encodes a 2.7 kb blastoderm-specific RNA. The primary structure of this RNA, and that of an overlapping 4.5 kb RNA, has been determined. The amino acid sequence of the predicted bsg25D protein has been compared to the NBRF protein database. Structural similarities between domains in the bsg25D, fos, and tropomyosin proteins, and their possible significance for early embryogenesis are discussed. Images PMID:3104878

  19. Quantitative imaging of gene expression in Drosophila embryos.

    PubMed

    Surkova, Svetlana; Myasnikova, Ekaterina; Kozlov, Konstantin N; Pisarev, Andrei; Reinitz, John; Samsonova, Maria

    2013-06-01

    Quantitative measurements derived using sophisticated microscopy techniques are essential for understanding the basic principles that control the behavior of biological systems. Here we describe a data pipeline developed to extract quantitative data on segmentation gene expression from confocal images of gene expression patterns in Drosophila. The pipeline consists of image segmentation, background removal, temporal characterization of an embryo, data registration, and data averaging. This pipeline has been successfully applied to obtain quantitative gene expression data at cellular resolution in space and at 6.5-min resolution in time. It has also enabled the construction of a spatiotemporal atlas of segmentation gene expression. We describe the software used to construct a workflow for extracting quantitative data on segmentation gene expression and the BREReA package, which implements the methods for background removal and registration of segmentation gene expression patterns. PMID:23734022

  20. Holocene left-slip rate determined by cosmogenic surface dating on the Xidatan segment of the Kunlun Fault (Qinghai, Chin

    SciTech Connect

    Guoguang, Z; Caffee, M; Finkel, R; G,; demer, Y; Meriaux, A S; Qunlu,; Ryerson, F J; Tapponnier, P; Van der Woerd, J

    1998-09-01

    Cosmogenic dating, using in-situ 26A1 and 10Be in quartz pebbles from alluvial terrace surfaces, constrains the late Holocene slip rate on the Xidatan segment of the Kuniun fault in northeastern Tibet. Two terrace risers offset by 24 ± 3 and 33 f± 4m, having respective ages of 1788 ± 388 and 2914 ± 471 yr, imply a slip rate of 12.1 ± 2.6 mm/yr. The full range of ages obtained ((less than or equal to) 22.8 k.y., most of them between 6.7 and 1.4 k.y.) confirm that terrace deposition and incision, hence landform evolution, are modulated by post-glacial climate change. Coupled with minimum offsets of 9-12 m, this slip rate implies that great earthquakes (M-8) with a recurrence time of 800-1000 yr. rupture the Kunlun fault n

  1. Material Parameter Determination of an L4-L5 Motion Segment Finite Element Model Under High Loading Rates.

    PubMed

    Pyles, C O; Zhang, J; Demetropoulos, C K; Bradfield, C A; Ott, K A; Armiger, R S; Merkle, A C

    2015-01-01

    Underbody blast (UBB) events impart vertical loads through a victim’s lumbar spine, resulting in fracture, paralysis, and disc rupture. Validated biofidelic lumbar models allow characterization of injury mechanisms and development of personal protective equipment. Previous studies have focused on lumbar mechanics under quasi-static loading. However, it is unclear how the role and response of individual spinal components of the lumbar spine change under dynamic loading. The present study leverages high-rate impacts of progressively dissected two-vertebra lumbar motion segments and Split-Hopkinson pressure bar tissue characterization to identify and validate material properties of a high-fidelity lumbar spine finite element model for UBB. The annulus fibrosus was modeled as a fiber-reinforced Mooney-Rivlin material, while ligaments were represented by nonlinear spring elements. Optimization and evaluation of material parameters was achieved by minimizing the root-mean-square (RMS) of compressive displacement and sagittal rotation for selected experimental conditions. Applying dynamic based material models and parameters resulted in a 0.42% difference between predicted and experiment axial compression during impact loading. This dynamically optimized lumbar model is suited for cross validation against whole-lumbar loading scenarios, and prediction of injury during UBB and other dynamic events. PMID:25996719

  2. Drosophila Blastorderm Analysis Software

    Energy Science and Technology Software Center (ESTSC)

    2006-10-25

    PointCloudMake analyzes 3D fluorescent images of whole Drosophila embryo and produces a table-style "PointCloud" file which contains the coordinates and volumes of all the nuclei, cells, their associated relative gene expression levels along with morphological features of the embryo. See: Luengo Hendrix et at 2006 3D Morphology and Gene Expression in the Drosophila Blastoderm at Cellular Resolution manuscript submitted LBNL # LBNL-60178 Knowles DW, Keranen SVE, Biggin M. Sudar S (2002) Mapping organism expression levelsmore » at cellular resolution in developing Drosophila. In: Conchello JA, Cogswell CJ, Wilson T, editors. Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX. pp. 57-64« less

  3. Identifying Benefit Segments among College Students.

    ERIC Educational Resources Information Center

    Brown, Joseph D.

    1991-01-01

    Using concept of market segmentation (dividing market into distinct groups requiring different product benefits), surveyed 398 college students to determine benefit segments among students selecting a college to attend and factors describing each benefit segment. Identified one major segment of students (classroomers) plus three minor segments…

  4. The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes.

    PubMed Central

    Spradling, A C; Stern, D; Beaton, A; Rhem, E J; Laverty, T; Mozden, N; Misra, S; Rubin, G M

    1999-01-01

    A fundamental goal of genetics and functional genomics is to identify and mutate every gene in model organisms such as Drosophila melanogaster. The Berkeley Drosophila Genome Project (BDGP) gene disruption project generates single P-element insertion strains that each mutate unique genomic open reading frames. Such strains strongly facilitate further genetic and molecular studies of the disrupted loci, but it has remained unclear if P elements can be used to mutate all Drosophila genes. We now report that the primary collection has grown to contain 1045 strains that disrupt more than 25% of the estimated 3600 Drosophila genes that are essential for adult viability. Of these P insertions, 67% have been verified by genetic tests to cause the associated recessive mutant phenotypes, and the validity of most of the remaining lines is predicted on statistical grounds. Sequences flanking >920 insertions have been determined to exactly position them in the genome and to identify 376 potentially affected transcripts from collections of EST sequences. Strains in the BDGP collection are available from the Bloomington Stock Center and have already assisted the research community in characterizing >250 Drosophila genes. The likely identity of 131 additional genes in the collection is reported here. Our results show that Drosophila genes have a wide range of sensitivity to inactivation by P elements, and provide a rationale for greatly expanding the BDGP primary collection based entirely on insertion site sequencing. We predict that this approach can bring >85% of all Drosophila open reading frames under experimental control. PMID:10471706

  5. Segmented combustor

    NASA Technical Reports Server (NTRS)

    Halila, Ely E. (Inventor)

    1994-01-01

    A combustor liner segment includes a panel having four sidewalls forming a rectangular outer perimeter. A plurality of integral supporting lugs are disposed substantially perpendicularly to the panel and extend from respective ones of the four sidewalls. A plurality of integral bosses are disposed substantially perpendicularly to the panel and extend from respective ones of the four sidewalls, with the bosses being shorter than the lugs. In one embodiment, the lugs extend through supporting holes in an annular frame for mounting the liner segments thereto, with the bosses abutting the frame for maintaining a predetermined spacing therefrom.

  6. Evidence for horizontal transfer of Wolbachia by a Drosophila mite.

    PubMed

    Brown, Amy N; Lloyd, Vett K

    2015-07-01

    Mites are common ectoparasites of Drosophila and have been implicated in bacterial and mobile element invasion of Drosophila stocks. The obligate endobacterium, Wolbachia, has widespread effects on gene expression in their arthropod hosts and alters host reproduction to enhance its survival and propagation, often with deleterious effects in Drosophila hosts. To determine whether Wolbachia could be transferred between Drosophila melanogaster laboratory stocks by the mite Tyrophagus putrescentiae, mites were introduced to Wolbachia-infected Drosophila vials. These vials were kept adjacent to mite-free and Wolbachia-uninfected Drosophila stock vials. The Wolbachia infection statuses of the infected and uninfected flies were checked from generation 1 to 5. Results indicate that Wolbachia DNA could be amplified from mites infesting Wolbachia-infected fly stocks and infection in the previously uninfected stocks arose within generation 1 or 2, concomitant with invasion of mites from the Wolbachia-infected stock. A possible mechanism for the transfer of Wolbachia from flies to mites and vice versa, can be inferred from time-lapse photography of fly and mite interactions. We demonstrated that mites ingest Drosophila corpses, including Wolbachia-infected corpses, and Drosophila larva ingest mites, providing possible sources of Wolbachia infection and transfer. This research demonstrated that T. putrescentiae white mites can facilitate Wolbachia transfer between Drosophila stocks and that this may occur by ingestion of infected corpses. Mite-vectored Wolbachia transfer allows for rapid establishment of Wolbachia infection within a new population. This mode of Wolbachia introduction may be relevant in nature as well as in the laboratory, and could have a variety of biological consequences. PMID:25921489

  7. A sensitive LC-MS/MS method for the simultaneous determination of amoxicillin and ambroxol in human plasma with segmental monitoring.

    PubMed

    Dong, Xin; Ding, Li; Cao, Xiaomei; Jiang, Liyuan; Zhong, Shuisheng

    2013-04-01

    Amoxicillin (AMO) degrades in plasma at room temperature and readily undergoes hydrolysis by the plasma amidase. In this paper, a novel, rapid and sensitive LC-MS/MS method operated in segmental and multiple reaction monitoring has been developed for the simultaneous determination of amoxicillin and ambroxol in human plasma. The degradation of amoxicillin in plasma was well prevented by immediate addition of 20 μL glacial acetic acid to 200 μL aliquot of freshly collected plasma samples before storage at -80°C. The sensitivity of the method was improved with segmental monitoring of the analytes, and lower limits of quantitation of 0.5 ng/mL for ambroxol and 5 ng/mL for amoxicillin were obtained. The sensitivity of our method was five times better than those of the existing methods. Furthermore, the mass response saturation problem with amoxicillin was avoided by diluting the deproteinized plasma samples with water before injection into the LC-MS/MS system. The method was successfully employed in a pharmacokinetic study of the compound amoxicillin and ambroxol hydrochloride tablets. PMID:23027506

  8. A Drosophila Model of Epidermolysis Bullosa Simplex.

    PubMed

    Bohnekamp, Jens; Cryderman, Diane E; Paululat, Achim; Baccam, Gabriel C; Wallrath, Lori L; Magin, Thomas M

    2015-08-01

    The blistering skin disorder epidermolysis bullosa simplex (EBS) results from dominant mutations in keratin 5 (K5) or keratin 14 (K14) genes, encoding the intermediate filament (IF) network of basal epidermal keratinocytes. The mechanisms governing keratin network formation and collapse due to EBS mutations remain incompletely understood. Drosophila lacks cytoplasmic IFs, providing a 'null' environment to examine the formation of keratin networks and determine mechanisms by which mutant keratins cause pathology. Here, we report that ubiquitous co-expression of transgenes encoding wild-type human K14 and K5 resulted in the formation of extensive keratin networks in Drosophila epithelial and non-epithelial tissues, causing no overt phenotype. Similar to mammalian cells, treatment of transgenic fly tissues with phosphatase inhibitors caused keratin network collapse, validating Drosophila as a genetic model system to investigate keratin dynamics. Co-expression of K5 and a K14(R125C) mutant that causes the most severe form of EBS resulted in widespread formation of EBS-like cytoplasmic keratin aggregates in epithelial and non-epithelial fly tissues. Expression of K14(R125C)/K5 caused semi-lethality; adult survivors developed wing blisters and were flightless due to a lack of intercellular adhesion during wing heart development. This Drosophila model of EBS is valuable for the identification of pathways altered by mutant keratins and for the development of EBS therapies. PMID:25830653

  9. The 5S genes of Drosophila melanogaster.

    PubMed

    Artavanis-Tsakonas, S; Schedl, P; Tschudi, C; Pirrotta, V; Steward, R; Gehring, W J

    1977-12-01

    We have cloned embryonic Drosophila DNA using the poly (dA-DT) connector method (Lobban and Kaiser, 1973) and the ampicillin-resistant plasmid pSF2124 (So, Gill and Falkow, 1975) as a cloning vehicle. Two clones, containing hybrid plasmids with sequences complementary to a 5S RNA probe isolated from Drosophila tissue culture cells, were identified by the Grunstein and Hogness (1975) colony hybridization procedure. One hybrid plasmid has a Drosophila insert which is comprised solely of tandem repeats of the 5S gene plus spacer sequences. The other plasmid contains an insert which has about 20 tandem 5S repeat units plus an additional 4 kilobases of adjacent sequences. The size of the 5S repeat unit was determined by gel electrophoresis and was found to be approximately 375 base pairs. We present a restriction map of both plasmids, and a detailed map of of the5S repeat unit. The 5S repat unit shows slight length and sequence heterogeneity. We present evidence suggesting that the 5S genes in Drosophila melanogaster may be arranged in a single continuous cluster. PMID:413625

  10. Corn leaf nitrate reductase - A nontoxic alternative to cadmium for photometric nitrate determinations in water samples by air-segmented continuous-flow analysis

    USGS Publications Warehouse

    Patton, C.J.; Fischer, A.E.; Campbell, W.H.; Campbell, E.R.

    2002-01-01

    Development, characterization, and operational details of an enzymatic, air-segmented continuous-flow analytical method for colorimetric determination of nitrate + nitrite in natural-water samples is described. This method is similar to U.S. Environmental Protection Agency method 353.2 and U.S. Geological Survey method 1-2545-90 except that nitrate is reduced to nitrite by soluble nitrate reductase (NaR, EC 1.6.6.1) purified from corn leaves rather than a packed-bed cadmium reactor. A three-channel, air-segmented continuous-flow analyzer-configured for simultaneous determination of nitrite (0.020-1.000 mg-N/L) and nitrate + nitrite (0.05-5.00 mg-N/L) by the nitrate reductase and cadmium reduction methods-was used to characterize analytical performance of the enzymatic reduction method. At a sampling rate of 90 h-1, sample interaction was less than 1% for all three methods. Method detection limits were 0.001 mg of NO2- -N/L for nitrite, 0.003 mg of NO3-+ NO2- -N/L for nitrate + nitrite by the cadmium-reduction method, and 0.006 mg of NO3- + NO2- -N/L for nitrate + nitrite by the enzymatic-reduction method. Reduction of nitrate to nitrite by both methods was greater than 95% complete over the entire calibration range. The difference between the means of nitrate + nitrite concentrations in 124 natural-water samples determined simultaneously by the two methods was not significantly different from zero at the p = 0.05 level.

  11. Heritable Endosymbionts of Drosophila

    PubMed Central

    Mateos, Mariana; Castrezana, Sergio J.; Nankivell, Becky J.; Estes, Anne M.; Markow, Therese A.; Moran, Nancy A.

    2006-01-01

    Although heritable microorganisms are increasingly recognized as widespread in insects, no systematic screens for such symbionts have been conducted in Drosophila species (the primary insect genetic models for studies of evolution, development, and innate immunity). Previous efforts screened relatively few Drosophila lineages, mainly for Wolbachia. We conducted an extensive survey of potentially heritable endosymbionts from any bacterial lineage via PCR screens of mature ovaries in 181 recently collected fly strains representing 35 species from 11 species groups. Due to our fly sampling methods, however, we are likely to have missed fly strains infected with sex ratio-distorting endosymbionts. Only Wolbachia and Spiroplasma, both widespread in insects, were confirmed as symbionts. These findings indicate that in contrast to some other insect groups, other heritable symbionts are uncommon in Drosophila species, possibly reflecting a robust innate immune response that eliminates many bacteria. A more extensive survey targeted these two symbiont types through diagnostic PCR in 1225 strains representing 225 species from 32 species groups. Of these, 19 species were infected by Wolbachia while only 3 species had Spiroplasma. Several new strains of Wolbachia and Spiroplasma were discovered, including ones divergent from any reported to date. The phylogenetic distribution of Wolbachia and Spiroplasma in Drosophila is discussed. PMID:16783009

  12. Method 366.0 Determination of Dissolved Silicate in Estuarine and Coastal Watersby Gas Segmented Continuous Flow Colorimetric Analysis

    EPA Science Inventory

    This method provides a procedure for the determination of dissolved silicate concentration in estuarine and coastal waters. The dissolved silicate is mainly in the form of silicic acid, H SiO , in estuarine and 4 4 coastal waters. All soluble silicate, including colloidal silici...

  13. Study of accuracy in the position determination with SALSA, a γ-scanning system for the characterization of segmented HPGe detectors

    NASA Astrophysics Data System (ADS)

    Hernandez-Prieto, A.; Quintana, B.; Martìn, S.; Domingo-Pardo, C.

    2016-07-01

    Accurate characterization of the electric response of segmented high-purity germanium (HPGe) detectors as a function of the interaction position is one of the current goals of the Nuclear Physics community seeking to perform γ-ray tracking or even imaging with these detectors. For this purpose, scanning devices must be developed to achieve the signal-position association with the highest precision. With a view to studying the accuracy achieved with SALSA, the SAlamanca Lyso-based Scanning Array, here we report a detailed study on the uncertainty sources and their effect in the position determination inside the HPGe detector to be scanned. The optimization performed on the design of SALSA, aimed at minimizing the effect of the uncertainty sources, afforded an intrinsic uncertainty of ∼2 mm for large coaxial detectors and ∼1 mm for planar ones.

  14. Pyrotechnic hazards classification and evaluation program. Phase 2, segment 3: Test plan for determining hazards associated with pyrotechnic manufacturing processes

    NASA Technical Reports Server (NTRS)

    1971-01-01

    A comprehensive test plan for determining the hazards associated with pyrotechnic manufacturing processes is presented. The rationale for each test is based on a systematic analysis of historical accounts of accidents and a detailed study of the characteristics of each manufacturing process. The most hazardous manufacturing operations have been determined to be pressing, mixing, reaming, and filling. The hazard potential of a given situation is evaluated in terms of the probabilities of initiation, communication, and transition to detonation (ICT). The characteristics which affect the ICT probabilities include the ignition mechanisms which are present either in normal or abnormal operation, the condition and properties of the pyrotechnic material, and the configuration of the processing equipment. Analytic expressions are derived which describe the physical conditions of the system, thus permitting a variety of processes to be evaluated in terms of a small number of experiments.

  15. The fruitfly Drosophila melanogaster contains a novel charged adipokinetic-hormone-family peptide.

    PubMed Central

    Schaffer, M H; Noyes, B E; Slaughter, C A; Thorne, G C; Gaskell, S J

    1990-01-01

    A member of the RPCH/AKH (red-pigment-concentrating hormone/adipokinetic hormone) family of arthropod neuropeptides was identified in the fruitfly Drosophila melanogaster, and its structure was determined by automated Edman degradation and m.s. using fast-atom-bombardment ionization and a tandem hybrid instrument capable of high sensitivity. The sequence of this peptide, which we call 'DAKH', is pGlu-Leu-Thr-Phe-Ser-Pro-Asp-Trp-NH2 (where pGlu is pyroglutamic acid and Trp-NH2 is tryptophan carboxyamide). H.p.l.c. analyses of extracts of the three body segments revealed that more than 80% of the peptide is contained in the thorax. Although DAKH is typical of family members in its general structure and distribution in the animal, it is unique in containing a residue which is charged under physiological conditions. The evolutionary significance of this change is considered. PMID:2117437

  16. The bacterial communities of Drosophila suzukii collected from undamaged cherries

    PubMed Central

    James, Pamela M.; Jospin, Guillaume; Lang, Jenna M.

    2014-01-01

    Drosophila suzukii is an introduced pest insect that feeds on undamaged, attached fruit. This diet is distinct from the fallen, discomposing fruits utilized by most other species of Drosophila. Since the bacterial microbiota of Drosophila, and of many other animals, is affected by diet, we hypothesized that the bacteria associated with D. suzukii are distinct from that of other Drosophila. Using 16S rDNA PCR and Illumina sequencing, we characterized the bacterial communities of larval and adult D. suzukii collected from undamaged, attached cherries in California, USA. We find that the bacterial communities associated with these samples of D. suzukii contain a high frequency of Tatumella. Gluconobacter and Acetobacter, two taxa with known associations with Drosophila, were also found, although at lower frequency than Tatumella in four of the five samples examined. Sampling D. suzukii from different locations and/or while feeding on different fruits is needed to determine the generality of the results determined by these samples. Nevertheless this is, to our knowledge, the first study characterizing the bacterial communities of this ecologically unique and economically important species of Drosophila. PMID:25101226

  17. R wave amplitude: a new determinant of failure of patients with coronary heart disease to manifest ST segment depression during exercise

    SciTech Connect

    Hakki, A.H.; Iskandrian, A.S.; Kutalek, S.; Hare, T.W.; Sokoloff, N.M.

    1984-05-01

    Patients with coronary artery disease may not manifest ST segment depression during exercise. Inadequate stress, mild coronary artery disease and severe left ventricular dysfunction have been postulated as mechanisms. The purpose of this study was to determine the influence of exercise R wave amplitude on ST segment depression in 81 patients with coronary artery disease (50% or greater diameter narrowing of one or more vessels). All patients underwent symptom-limited treadmill exercise testing and 71 patients (88%) had concomitant thallium-201 imaging. In 26 patients, the exercise R wave amplitude in electrocardiographic lead V5 was less than 11 mm (Group I), and in 55 patients it was 11 mm or greater (Group II). The two groups were similar with regard to age, sex, propranolol administration and left ventricular function. There was a significant difference in the incidence of positive exercise electrocardiograms in the two groups (2 patients (8%) in Group I and 27 patients (49%) in Group II; p . 0.002), despite similar exercise heart rate and extent of coronary artery disease. Myocardial ischemia, manifested by exercise-induced angina or exercise-induced thallium-201 perfusion defects, was similar in both groups. Thallium-201 imaging showed perfusion defects in 73% of patients in Group I and in 76% of patients in Group II (p . not significant). Thus, R wave amplitude is a new determinant of failure to develop ST depression during exercise. A low R wave amplitude (less than 11 mm) is rarely associated with ST depression, even in patients with multivessel coronary artery disease. Exercise thallium-201 imaging is a valuable diagnostic tool in patients with low R wave amplitude.

  18. The FHA domain determines Drosophila Chk2/Mnk localization to key mitotic structures and is essential for early embryonic DNA damage responses.

    PubMed

    Takada, Saeko; Collins, Eric R; Kurahashi, Kayo

    2015-05-15

    DNA damage responses, including mitotic centrosome inactivation, cell-cycle delay in mitosis, and nuclear dropping from embryo cortex, maintain genome integrity in syncytial Drosophila embryos. A conserved signaling kinase, Chk2, known as Mnk/Loki, is essential for the responses. Here we demonstrate that functional EGFP-Mnk expressed from a transgene localizes to the nucleus, centrosomes, interkinetochore/centromere region, midbody, and pseudocleavage furrows without DNA damage and in addition forms numerous foci/aggregates on mitotic chromosomes upon DNA damage. We expressed EGFP-tagged Mnk deletion or point mutation variants and investigated domain functions of Mnk in vivo. A triple mutation in the phosphopeptide-binding site of the forkhead-associated (FHA) domain disrupted normal Mnk localization except to the nucleus. The mutation also disrupted Mnk foci formation on chromosomes upon DNA damage. FHA mutations and deletion of the SQ/TQ-cluster domain (SCD) abolished Mnk transphosphorylations and autophosphorylations, indicative of kinase activation after DNA damage. A potent NLS was found at the C-terminus, which is required for normal Mnk function. We propose that the FHA domain in Mnk plays essential dual functions in mediating embryonic DNA damage responses by means of its phosphopeptide-binding ability: activating Mnk in the nucleus upon DNA damage and recruiting Mnk to multiple subcellular structures independently of DNA damage. PMID:25808488

  19. The FHA domain determines Drosophila Chk2/Mnk localization to key mitotic structures and is essential for early embryonic DNA damage responses

    PubMed Central

    Takada, Saeko; Collins, Eric R.; Kurahashi, Kayo

    2015-01-01

    DNA damage responses, including mitotic centrosome inactivation, cell-cycle delay in mitosis, and nuclear dropping from embryo cortex, maintain genome integrity in syncytial Drosophila embryos. A conserved signaling kinase, Chk2, known as Mnk/Loki, is essential for the responses. Here we demonstrate that functional EGFP-Mnk expressed from a transgene localizes to the nucleus, centrosomes, interkinetochore/centromere region, midbody, and pseudocleavage furrows without DNA damage and in addition forms numerous foci/aggregates on mitotic chromosomes upon DNA damage. We expressed EGFP-tagged Mnk deletion or point mutation variants and investigated domain functions of Mnk in vivo. A triple mutation in the phosphopeptide-binding site of the forkhead-associated (FHA) domain disrupted normal Mnk localization except to the nucleus. The mutation also disrupted Mnk foci formation on chromosomes upon DNA damage. FHA mutations and deletion of the SQ/TQ-cluster domain (SCD) abolished Mnk transphosphorylations and autophosphorylations, indicative of kinase activation after DNA damage. A potent NLS was found at the C-terminus, which is required for normal Mnk function. We propose that the FHA domain in Mnk plays essential dual functions in mediating embryonic DNA damage responses by means of its phosphopeptide-binding ability: activating Mnk in the nucleus upon DNA damage and recruiting Mnk to multiple subcellular structures independently of DNA damage. PMID:25808488

  20. [Segmental neurofibromatosis].

    PubMed

    Zulaica, A; Peteiro, C; Pereiro, M; Pereiro Ferreiros, M; Quintas, C; Toribio, J

    1989-01-01

    Four cases of segmental neurofibromatosis (SNF) are reported. It is a rare entity considered to be a localized variant of neurofibromatosis (NF)-Riccardi's type V. Two cases are male and two female. The lesions are located to the head in a patient and the other three cases in the trunk. No family history nor transmission to progeny were manifested. The rest of the organs are undamaged. PMID:2502696

  1. LUSH odorant-binding protein mediates chemosensory responses to alcohols in Drosophila melanogaster.

    PubMed Central

    Kim, M S; Repp, A; Smith, D P

    1998-01-01

    The molecular mechanisms mediating chemosensory discrimination in insects are unknown. Using the enhancer trapping approach, we identified a new Drosophila mutant, lush, with odorant-specific defects in olfactory behavior. lush mutant flies are abnormally attracted to high concentrations of ethanol, propanol, and butanol but have normal chemosensory responses to other odorants. We show that wild-type flies have an active olfactory avoidance mechanism to prevent attraction to concentrated alcohol, and this response is defective in lush mutants. This suggests that the defective olfactory behavior associated with the lush mutation may result from a specific defect in chemoavoidance. lush mutants have a 3-kb deletion that produces a null allele of a new member of the invertebrate odorant-binding protein family, LUSH. LUSH is normally expressed exclusively in a subset of trichoid chemosensory sensilla located on the ventral-lateral surface of the third antennal segment. LUSH is secreted from nonneuronal support cells into the sensillum lymph that bathes the olfactory neurons within these sensilla. Reintroduction of a cloned wild-type copy of lush into the mutant background completely restores wild-type olfactory behavior, demonstrating that this odorant-binding protein is required in a subset of sensilla for normal chemosensory behavior to a subset of odorants. These findings provide direct evidence that odorant-binding proteins are required for normal chemosensory behavior in Drosophila and may partially determine the chemical specificity of olfactory neurons in vivo. PMID:9755202

  2. Effect of inversion polymorphism on the neutral nucleotide variability of linked chromosomal regions in Drosophila.

    PubMed Central

    Navarro, A; Barbadilla, A; Ruiz, A

    2000-01-01

    Recombination is a main factor determining nucleotide variability in different regions of the genome. Chromosomal inversions, which are ubiquitous in the genus Drosophila, are known to reduce and redistribute recombination, and thus their specific effect on nucleotide variation may be of major importance as an explanatory factor for levels of DNA variation. Here, we use the coalescent approach to study this effect. First, we develop analytical expressions to predict nucleotide variability in old inversion polymorphisms that have reached mutation-drift-flux equilibrium. The effects on nucleotide variability of a new arrangement appearing in the population and reaching a stable polymorphism are then studied by computer simulation. We show that inversions modulate nucleotide variability in a complex way. The establishment of an inversion polymorphism involves a partial selective sweep that eliminates part of the variability in the population. This is followed by a slow convergence to the equilibrium values. During this convergence, regions close to the breakpoints exhibit much lower variability than central regions. However, at equilibrium, regions close to the breakpoints have higher levels of variability and differentiation between arrangements than regions in the middle of the inverted segment. The implications of these findings for overall variability levels during the evolution of Drosophila species are discussed. PMID:10835391

  3. Aging Studies in Drosophila melanogaster

    PubMed Central

    Sun, Yaning; Yolitz, Jason; Wang, Cecilia; Spangler, Edward; Zhan, Ming; Zou, Sige

    2015-01-01

    Summary Drosophila is a genetically tractable system ideal for investigating the mechanisms of aging and developing interventions for promoting healthy aging. Here we describe methods commonly used in Drosophila aging research. These include basic approaches for preparation of diets and measurements of lifespan, food intake and reproductive output. We also describe some commonly used assays to measure changes in physiological and behavioral functions of Drosophila in aging, such as stress resistance and locomotor activity. PMID:23929099

  4. The use of segmented cathodes to determine the spoke current density distribution in high power impulse magnetron sputtering plasmas

    SciTech Connect

    Poolcharuansin, Phitsanu; Estrin, Francis Lockwood; Bradley, James W.

    2015-04-28

    The localized target current density associated with quasi-periodic ionization zones (spokes) has been measured in a high power impulse magnetron sputtering (HiPIMS) discharge using an array of azimuthally separated and electrical isolated probes incorporated into a circular aluminum target. For a particular range of operating conditions (pulse energies up to 2.2 J and argon pressures from 0.2 to 1.9 Pa), strong oscillations in the probe current density are seen with amplitudes up to 52% above a base value. These perturbations, identified as spokes, travel around the discharge above the target in the E×B direction. Using phase information from the angularly separated probes, the spoke drift speeds, angular frequencies, and mode number have been determined. Generally, at low HiPIMS pulse energies E{sub p} < 0.8 J, spokes appear to be chaotic in nature (with random arrival times), however as E{sub p} increases, coherent spokes are observed with velocities between 6.5 and 10 km s{sup −1} and mode numbers m = 3 or above. At E{sub p} > 1.8 J, the plasma becomes spoke-free. The boundaries between chaotic, coherent, and no-spoke regions are weakly dependent on pressure. During each HiPIMS pulse, the spoke velocities increase by about 50%. Such an observation is explained by considering spoke velocities to be determined by the critical ionization velocity, which changes as the plasma composition changes during the pulse. From the shape of individual current density oscillations, it appears that the leading edge of the spoke is associated with a slow increase in local current density to the target and the rear with a more rapid decrease. The measurements show that the discharge current density associated with individual spokes is broadly spread over a wide region of the target.

  5. Rediscovering market segmentation.

    PubMed

    Yankelovich, Daniel; Meer, David

    2006-02-01

    In 1964, Daniel Yankelovich introduced in the pages of HBR the concept of nondemographic segmentation, by which he meant the classification of consumers according to criteria other than age, residence, income, and such. The predictive power of marketing studies based on demographics was no longer strong enough to serve as a basis for marketing strategy, he argued. Buying patterns had become far better guides to consumers' future purchases. In addition, properly constructed nondemographic segmentations could help companies determine which products to develop, which distribution channels to sell them in, how much to charge for them, and how to advertise them. But more than 40 years later, nondemographic segmentation has become just as unenlightening as demographic segmentation had been. Today, the technique is used almost exclusively to fulfill the needs of advertising, which it serves mainly by populating commercials with characters that viewers can identify with. It is true that psychographic types like "High-Tech Harry" and "Joe Six-Pack" may capture some truth about real people's lifestyles, attitudes, self-image, and aspirations. But they are no better than demographics at predicting purchase behavior. Thus they give corporate decision makers very little idea of how to keep customers or capture new ones. Now, Daniel Yankelovich returns to these pages, with consultant David Meer, to argue the case for a broad view of nondemographic segmentation. They describe the elements of a smart segmentation strategy, explaining how segmentations meant to strengthen brand identity differ from those capable of telling a company which markets it should enter and what goods to make. And they introduce their "gravity of decision spectrum", a tool that focuses on the form of consumer behavior that should be of the greatest interest to marketers--the importance that consumers place on a product or product category. PMID:16485810

  6. Fault rupture segmentation

    NASA Astrophysics Data System (ADS)

    Cleveland, Kenneth Michael

    A critical foundation to earthquake study and hazard assessment is the understanding of controls on fault rupture, including segmentation. Key challenges to understanding fault rupture segmentation include, but are not limited to: What determines if a fault segment will rupture in a single great event or multiple moderate events? How is slip along a fault partitioned between seismic and seismic components? How does the seismicity of a fault segment evolve over time? How representative are past events for assessing future seismic hazards? In order to address the difficult questions regarding fault rupture segmentation, new methods must be developed that utilize the information available. Much of the research presented in this study focuses on the development of new methods for attacking the challenges of understanding fault rupture segmentation. Not only do these methods exploit a broader band of information within the waveform than has traditionally been used, but they also lend themselves to the inclusion of even more seismic phases providing deeper understandings. Additionally, these methods are designed to be fast and efficient with large datasets, allowing them to utilize the enormous volume of data available. Key findings from this body of work include demonstration that focus on fundamental earthquake properties on regional scales can provide general understanding of fault rupture segmentation. We present a more modern, waveform-based method that locates events using cross-correlation of the Rayleigh waves. Additionally, cross-correlation values can also be used to calculate precise earthquake magnitudes. Finally, insight regarding earthquake rupture directivity can be easily and quickly exploited using cross-correlation of surface waves.

  7. DNA G-segment bending is not the sole determinant of topology simplification by type II DNA topoisomerases.

    PubMed

    Thomson, Neil H; Santos, Sergio; Mitchenall, Lesley A; Stuchinskaya, Tanya; Taylor, James A; Maxwell, Anthony

    2014-01-01

    DNA topoisomerases control the topology of DNA. Type II topoisomerases exhibit topology simplification, whereby products of their reactions are simplified beyond that expected based on thermodynamic equilibrium. The molecular basis for this process is unknown, although DNA bending has been implicated. To investigate the role of bending in topology simplification, the DNA bend angles of four enzymes of different types (IIA and IIB) were measured using atomic force microscopy (AFM). The enzymes tested were Escherichia coli topo IV and yeast topo II (type IIA enzymes that exhibit topology simplification), and Methanosarcina mazei topo VI and Sulfolobus shibatae topo VI (type IIB enzymes, which do not). Bend angles were measured using the manual tangent method from topographical AFM images taken with a novel amplitude-modulated imaging mode: small amplitude small set-point (SASS), which optimises resolution for a given AFM tip size and minimises tip convolution with the sample. This gave improved accuracy and reliability and revealed that all 4 topoisomerases bend DNA by a similar amount: ~120° between the DNA entering and exiting the enzyme complex. These data indicate that DNA bending alone is insufficient to explain topology simplification and that the 'exit gate' may be an important determinant of this process. PMID:25142513

  8. Volumetric Semantic Segmentation using Pyramid Context Features

    PubMed Central

    Barron, Jonathan T.; Arbeláez, Pablo; Keränen, Soile V. E.; Biggin, Mark D.; Knowles, David W.; Malik, Jitendra

    2015-01-01

    We present an algorithm for the per-voxel semantic segmentation of a three-dimensional volume. At the core of our algorithm is a novel “pyramid context” feature, a descriptive representation designed such that exact per-voxel linear classification can be made extremely efficient. This feature not only allows for efficient semantic segmentation but enables other aspects of our algorithm, such as novel learned features and a stacked architecture that can reason about self-consistency. We demonstrate our technique on 3D fluorescence microscopy data of Drosophila embryos for which we are able to produce extremely accurate semantic segmentations in a matter of minutes, and for which other algorithms fail due to the size and high-dimensionality of the data, or due to the difficulty of the task. PMID:26029008

  9. Premating isolation is determined by larval rearing substrates in cactophilic Drosophila mojavensis. X. Age-specific dynamics of adult epicuticular hydrocarbon expression in response to different host plants

    PubMed Central

    Etges, William J; de Oliveira, Cassia C

    2014-01-01

    Analysis of sexual selection and sexual isolation in Drosophila mojavensis and its relatives has revealed a pervasive role of rearing substrates on adult courtship behavior when flies were reared on fermenting cactus in preadult stages. Here, we assessed expression of contact pheromones comprised of epicuticular hydrocarbons (CHCs) from eclosion to 28 days of age in adults from two populations reared on fermenting tissues of two host cacti over the entire life cycle. Flies were never exposed to laboratory food and showed significant reductions in average CHC amounts consistent with CHCs of wild-caught flies. Overall, total hydrocarbon amounts increased from eclosion to 14–18 days, well past age at sexual maturity, and then declined in older flies. Most flies did not survive past 4 weeks. Baja California and mainland populations showed significantly different age-specific CHC profiles where Baja adults showed far less age-specific changes in CHC expression. Adults from populations reared on the host cactus typically used in nature expressed more CHCs than on the alternate host. MANCOVA with age as the covariate for the first six CHC principal components showed extensive differences in CHC composition due to age, population, cactus, sex, and age × population, age × sex, and age × cactus interactions. Thus, understanding variation in CHC composition as adult D. mojavensis age requires information about population and host plant differences, with potential influences on patterns of mate choice, sexual selection, and sexual isolation, and ultimately how these pheromones are expressed in natural populations. Studies of drosophilid aging in the wild are badly needed. PMID:25360246

  10. Developmental roles of 21 Drosophila transcription factors are determined by quantitative differences in binding to an overlapping set of thousands of genomic regions

    SciTech Connect

    MacArthur, Stewart; Li, Xiao-Yong; Li, Jingyi; Brown, James B.; Chu, Hou Cheng; Zeng, Lucy; Grondona, Brandi P.; Hechmer, Aaron; Simirenko, Lisa; Keranen, Soile V.E.; Knowles, David W.; Stapleton, Mark; Bickel, Peter; Biggin, Mark D.; Eisen, Michael B.

    2009-05-15

    BACKGROUND: We previously established that six sequence-specific transcription factors that initiate anterior/posterior patterning in Drosophila bind to overlapping sets of thousands of genomic regions in blastoderm embryos. While regions bound at high levels include known and probable functional targets, more poorly bound regions are preferentially associated with housekeeping genes and/or genes not transcribed in the blastoderm, and are frequently found in protein coding sequences or in less conserved non-coding DNA, suggesting that many are likely non-functional. RESULTS: Here we show that an additional 15 transcription factors that regulate other aspects of embryo patterning show a similar quantitative continuum of function and binding to thousands of genomic regions in vivo. Collectively, the 21 regulators show a surprisingly high overlap in the regions they bind given that they belong to 11 DNA binding domain families, specify distinct developmental fates, and can act via different cis-regulatory modules. We demonstrate, however, that quantitative differences in relative levels of binding to shared targets correlate with the known biological and transcriptional regulatory specificities of these factors. CONCLUSIONS: It is likely that the overlap in binding of biochemically and functionally unrelated transcription factors arises from the high concentrations of these proteins in nuclei, which, coupled with their broad DNA binding specificities, directs them to regions of open chromatin. We suggest that most animal transcription factors will be found to show a similar broad overlapping pattern of binding in vivo, with specificity achieved by modulating the amount, rather than the identity, of bound factor.

  11. Drosophila by the dozen

    SciTech Connect

    Celniker, Susan E.; Hoskins, Roger A.

    2007-07-13

    This year's conference on Drosophila research illustratedwell the current focus of Drosophila genomics on the comprehensiveidentification of functional elements in the genome sequence, includingmRNA transcripts arising from multiple alternative start sites and splicesites, a multiplicity of noncoding transcripts and small RNAs,identification of binding sites for transcription factors, sequenceconservation in related species and sequence variation within species.Resources and technologies for genetics and functional genomics aresteadily being improved, including the building of collections oftransposon insertion mutants and hairpin constructs for RNA interference(RNAi). The conference also highlighted progress in the use of genomicinformation by many laboratories to study diverse aspects of biology andmodels of human disease. Here we will review a few highlights of especialinterest to readers of Genome Biology.

  12. The Drosophila Auditory System

    PubMed Central

    Boekhoff-Falk, Grace; Eberl, Daniel F.

    2013-01-01

    Development of a functional auditory system in Drosophila requires specification and differentiation of the chordotonal sensilla of Johnston’s organ (JO) in the antenna, correct axonal targeting to the antennal mechanosensory and motor center (AMMC) in the brain, and synaptic connections to neurons in the downstream circuit. Chordotonal development in JO is functionally complicated by structural, molecular and functional diversity that is not yet fully understood, and construction of the auditory neural circuitry is only beginning to unfold. Here we describe our current understanding of developmental and molecular mechanisms that generate the exquisite functions of the Drosophila auditory system, emphasizing recent progress and highlighting important new questions arising from research on this remarkable sensory system. PMID:24719289

  13. Segmental neurofibromatosis.

    PubMed

    Sobjanek, Michał; Dobosz-Kawałko, Magdalena; Michajłowski, Igor; Pęksa, Rafał; Nowicki, Roman

    2014-12-01

    Segmental neurofibromatosis or type V neurofibromatosis is a rare genodermatosis characterized by neurofibromas, café-au-lait spots and neurofibromas limited to a circumscribed body region. The disease may be associated with systemic involvement and malignancies. The disorder has not been reported yet in the Polish medical literature. A 63-year-old Caucasian woman presented with a 20-year history of multiple, flesh colored, dome-shaped, soft to firm nodules situated in the right lumbar region. A histopathologic evaluation of three excised tumors revealed neurofibromas. No neurological and ophthalmologic symptoms of neurofibromatosis were diagnosed. PMID:25610358

  14. Segmental neurofibromatosis.

    PubMed

    Adigun, Chris G; Stein, Jennifer

    2011-01-01

    A 59-year-old man presented for evaluation and excision of non-tender, fleshy nodules that were arranged in a dermatomal distribution from the left side of the chest to the left axilla. A biopsy specimen of a nodule was consistent with a neurofibroma. Owing to the lack of other cutaneous findings, the lack of a family history of neurofibromatosis, and the dermatomal distribution of the neurofibromas, this patient met the criteria for a diagnosis of segmental neurofibromatosis (SNF) according to Riccardi's definition of SNF and classification of neurofibromatosis. Because the patient has no complications of neurofibromatosis 1 no medical treatment is required. PMID:22031651

  15. Segmental neurofibromatosis

    PubMed Central

    Dobosz-Kawałko, Magdalena; Michajłowski, Igor; Pęksa, Rafał; Nowicki, Roman

    2014-01-01

    Segmental neurofibromatosis or type V neurofibromatosis is a rare genodermatosis characterized by neurofibromas, café-au-lait spots and neurofibromas limited to a circumscribed body region. The disease may be associated with systemic involvement and malignancies. The disorder has not been reported yet in the Polish medical literature. A 63-year-old Caucasian woman presented with a 20-year history of multiple, flesh colored, dome-shaped, soft to firm nodules situated in the right lumbar region. A histopathologic evaluation of three excised tumors revealed neurofibromas. No neurological and ophthalmologic symptoms of neurofibromatosis were diagnosed. PMID:25610358

  16. Uniqueness in the determination of vibration sources in rectangular Germain-Lagrange plates using displacement measurements over line segments with arbitrary small length

    NASA Astrophysics Data System (ADS)

    Kawano, Alexandre

    2013-08-01

    The theme of this work is related to the field of vibration and source detection, which is important in naval, aerospace and civil engineering industries. The detection of unexpected vibration sources, in general, signals malfunctioning, or even an undesired presence in the case of defense systems. The focus will be on thin plates, which are among the basic building blocks of large complex structures. Here, we consider loads acting on a rectangular plate R of the product form g(t)Q(x), where the function of time g has a continuous first derivative and the spatial load distribution Q is a square-integrable function over R. We prove that the observation of the displacement of a line segment with arbitrary length parallel to one of the sides of the plate is enough for the determination of Q, provided that the interval of time is long enough. We also prove that the normal derivative along a side of the rectangle measured for an arbitrarily small interval of time is sufficient to determine the spatial load distribution Q. The method used to obtain the results is based on the series decomposition of the dynamic response and an analysis of the almost periodic distribution that arises from it.

  17. Optimal segmentation and packaging process

    DOEpatents

    Kostelnik, Kevin M.; Meservey, Richard H.; Landon, Mark D.

    1999-01-01

    A process for improving packaging efficiency uses three dimensional, computer simulated models with various optimization algorithms to determine the optimal segmentation process and packaging configurations based on constraints including container limitations. The present invention is applied to a process for decontaminating, decommissioning (D&D), and remediating a nuclear facility involving the segmentation and packaging of contaminated items in waste containers in order to minimize the number of cuts, maximize packaging density, and reduce worker radiation exposure. A three-dimensional, computer simulated, facility model of the contaminated items are created. The contaminated items are differentiated. The optimal location, orientation and sequence of the segmentation and packaging of the contaminated items is determined using the simulated model, the algorithms, and various constraints including container limitations. The cut locations and orientations are transposed to the simulated model. The contaminated items are actually segmented and packaged. The segmentation and packaging may be simulated beforehand. In addition, the contaminated items may be cataloged and recorded.

  18. Effect of sterol metabolism in the yeast-Drosophila system on the frequency of radiation-induced aneuploidy in the Drosophila melanogaster oocytes

    SciTech Connect

    Savitskii, V.V.; Luchnikova, E.M.; Inge-Vechtomov, S.G.

    1986-01-01

    The effect of sterol metabolism on induced mutagenesis of Drosophila melanogaster was studied in the ecogenetic system of yeast-Drosophila. Sterol deficiency was created in Drosophila by using the biomass of live cells of Saccharomyces cerevisiae strain 9-2-P712 till mutation in locus nys/sup r1/ blocking the synthesis of ergosterol as the food. It was found that rearing of Drosophila females on the mutant yeast increases the frequency of loss and nondisjunction of X chromosomes induced in mature oocytes by X rays (1000 R). Addition of 0.1% of cholesterol solution in 10% ethanol to the yeast biomass restores the resistance of oocyte to X irradiation to the control level. The possible hormonal effect on membrane leading to increased radiation-induced aneuploidy in Drosophila and the role of sterol metabolism in determining the resistance to various damaging factors are discussed.

  19. The genome sequence of Drosophila melanogaster.

    SciTech Connect

    2000-03-24

    The fly Drosophila melanogaster is one of the most intensively studied organisms in biology and serves as a model system for the investigation of many developmental and cellular processes common to higher eukaryotes, including humans. We have determined the nucleotide sequence of nearly all of the {approximately}120-megabase euchromatic portion of the Drosophila genome using a whole-genome shotgun sequencing strategy supported by extensive clone-based sequence and a high-quality bacterial artificial chromosome physical map. Efforts are under way to close the remaining gaps; however, the sequence is of sufficient accuracy and contiguity to be declared substantially complete and to support an initial analysis of genome structure and preliminary gene annotation and interpretation. The genome encodes {approximately}13,600 genes, somewhat fewer than the smaller Caenorhabditis elegans genome, but with comparable functional diversity.

  20. dachshund Potentiates Hedgehog Signaling during Drosophila Retinogenesis

    PubMed Central

    Aerts, Stein; Casares, Fernando; Janody, Florence

    2016-01-01

    Proper organ patterning depends on a tight coordination between cell proliferation and differentiation. The patterning of Drosophila retina occurs both very fast and with high precision. This process is driven by the dynamic changes in signaling activity of the conserved Hedgehog (Hh) pathway, which coordinates cell fate determination, cell cycle and tissue morphogenesis. Here we show that during Drosophila retinogenesis, the retinal determination gene dachshund (dac) is not only a target of the Hh signaling pathway, but is also a modulator of its activity. Using developmental genetics techniques, we demonstrate that dac enhances Hh signaling by promoting the accumulation of the Gli transcription factor Cubitus interruptus (Ci) parallel to or downstream of fused. In the absence of dac, all Hh-mediated events associated to the morphogenetic furrow are delayed. One of the consequences is that, posterior to the furrow, dac- cells cannot activate a Roadkill-Cullin3 negative feedback loop that attenuates Hh signaling and which is necessary for retinal cells to continue normal differentiation. Therefore, dac is part of an essential positive feedback loop in the Hh pathway, guaranteeing the speed and the accuracy of Drosophila retinogenesis. PMID:27442438

  1. Compatibility of segmented thermoelectric generators

    NASA Technical Reports Server (NTRS)

    Snyder, J.; Ursell, T.

    2002-01-01

    It is well known that power generation efficiency improves when materials with appropriate properties are combined either in a cascaded or segmented fashion across a temperature gradient. Past methods for determining materials used in segmentation weremainly concerned with materials that have the highest figure of merit in the temperature range. However, the example of SiGe segmented with Bi2Te3 and/or various skutterudites shows a marked decline in device efficiency even though SiGe has the highest figure of merit in the temperature range. The origin of the incompatibility of SiGe with other thermoelectric materials leads to a general definition of compatibility and intrinsic efficiency. The compatibility factor derived as = (Jl+zr - 1) a is a function of only intrinsic material properties and temperature, which is represented by a ratio of current to conduction heat. For maximum efficiency the compatibility factor should not change with temperature both within a single material, and in the segmented leg as a whole. This leads to a measure of compatibility not only between segments, but also within a segment. General temperature trends show that materials are more self compatible at higher temperatures, and segmentation is more difficult across a larger -T. The compatibility factor can be used as a quantitative guide for deciding whether a material is better suited for segmentation orcascading. Analysis of compatibility factors and intrinsic efficiency for optimal segmentation are discussed, with intent to predict optimal material properties, temperature interfaces, and/or currentheat ratios.

  2. Short, long, and beyond: molecular and embryological approaches to insect segmentation.

    PubMed

    Davis, Gregory K; Patel, Nipam H

    2002-01-01

    Over the past dozen years, studies comparing the expression of orthologues of the Drosophila segmentation genes among various insects have served to broaden our view of the ways in which insects make segments. The molecular data suggest that, although the overall genetic mechanisms of segmentation during embryogenesis have been conserved, the details of this process vary both within and between various insect orders. Here we summarize comparative gene expression data relevant to segmentation with an emphasis on understanding the extent of molecular patterning prior to gastrulation. These results are discussed in embryological context with an eye toward understanding the evolution of segmentation within insects. PMID:11729088

  3. Genetic control of Drosophila nerve cord development

    NASA Technical Reports Server (NTRS)

    Skeath, James B.; Thor, Stefan

    2003-01-01

    The Drosophila ventral nerve cord has been a central model system for studying the molecular genetic mechanisms that control CNS development. Studies show that the generation of neural diversity is a multistep process initiated by the patterning and segmentation of the neuroectoderm. These events act together with the process of lateral inhibition to generate precursor cells (neuroblasts) with specific identities, distinguished by the expression of unique combinations of regulatory genes. The expression of these genes in a given neuroblast restricts the fate of its progeny, by activating specific combinations of downstream genes. These genes in turn specify the identity of any given postmitotic cell, which is evident by its cellular morphology and choice of neurotransmitter.

  4. Hedgehog signaling pathway function conserved in Tribolium segmentation.

    PubMed

    Farzana, Laila; Brown, Susan J

    2008-04-01

    In Drosophila, maintenance of parasegmental boundaries and formation of segmental grooves depend on interactions between segment polarity genes. Wingless and Engrailed appear to have similar roles in both short and long germ segmentation, but relatively little is known about the extent to which Hedgehog signaling is conserved. In a companion study to the Tribolium genome project, we analyzed the expression and function of hedgehog, smoothened, patched, and cubitus interruptus orthologs during segmentation in Tribolium. Their expression was largely conserved between Drosophila and Tribolium. Parental RNAi analysis of positive regulators of the pathway (Tc-hh, Tc-smo, or Tc-ci) resulted in small spherical cuticles with little or no evidence of segmental grooves. Segmental Engrailed expression in these embryos was initiated but not maintained. Wingless-independent Engrailed expression in the CNS was maintained and became highly compacted during germ band retraction, providing evidence that derivatives from every segment were present in these small spherical embryos. On the other hand, RNAi analysis of a negative regulator (Tc-ptc) resulted in embryos with ectopic segmental grooves visible during germband elongation but not discernible in the first instar larval cuticles. These transient grooves formed adjacent to Engrailed expressing cells that encircled wider than normal wg domains in the Tc-ptc RNAi embryos. These results suggest that the en-wg-hh gene circuit is functionally conserved in the maintenance of segmental boundaries during germ band retraction and groove formation in Tribolium and that the segment polarity genes form a robust genetic regulatory module in the segmentation of this short germ insect. PMID:18392879

  5. Quantitative analysis of gene function in the Drosophila embryo.

    PubMed Central

    Tracey, W D; Ning, X; Klingler, M; Kramer, S G; Gergen, J P

    2000-01-01

    The specific functions of gene products frequently depend on the developmental context in which they are expressed. Thus, studies on gene function will benefit from systems that allow for manipulation of gene expression within model systems where the developmental context is well defined. Here we describe a system that allows for genetically controlled overexpression of any gene of interest under normal physiological conditions in the early Drosophila embryo. This regulated expression is achieved through the use of Drosophila lines that express a maternal mRNA for the yeast transcription factor GAL4. Embryos derived from females that express GAL4 maternally activate GAL4-dependent UAS transgenes at uniform levels throughout the embryo during the blastoderm stage of embryogenesis. The expression levels can be quantitatively manipulated through the use of lines that have different levels of maternal GAL4 activity. Specific phenotypes are produced by expression of a number of different developmental regulators with this system, including genes that normally do not function during Drosophila embryogenesis. Analysis of the response to overexpression of runt provides evidence that this pair-rule segmentation gene has a direct role in repressing transcription of the segment-polarity gene engrailed. The maternal GAL4 system will have applications both for the measurement of gene activity in reverse genetic experiments as well as for the identification of genetic factors that have quantitative effects on gene function in vivo. PMID:10628987

  6. Gene expression profiles uncover individual identities of gnathal neuroblasts and serial homologies in the embryonic CNS of Drosophila

    PubMed Central

    Urbach, Rolf; Jussen, David; Technau, Gerhard M.

    2016-01-01

    The numbers and types of progeny cells generated by neural stem cells in the developing CNS are adapted to its region-specific functional requirements. In Drosophila, segmental units of the CNS develop from well-defined patterns of neuroblasts. Here we constructed comprehensive neuroblast maps for the three gnathal head segments. Based on the spatiotemporal pattern of neuroblast formation and the expression profiles of 46 marker genes (41 transcription factors), each neuroblast can be uniquely identified. Compared with the thoracic ground state, neuroblast numbers are progressively reduced in labial, maxillary and mandibular segments due to smaller sizes of neuroectodermal anlagen and, partially, to suppression of neuroblast formation and induction of programmed cell death by the Hox gene Deformed. Neuroblast patterns are further influenced by segmental modifications in dorsoventral and proneural gene expression. With the previously published neuroblast maps and those presented here for the gnathal region, all neuroectodermal neuroblasts building the CNS of the fly (ventral nerve cord and brain, except optic lobes) are now individually identified (in total 2×567 neuroblasts). This allows, for the first time, a comparison of the characteristics of segmental populations of stem cells and to screen for serially homologous neuroblasts throughout the CNS. We show that approximately half of the deutocerebral and all of the tritocerebral (posterior brain) and gnathal neuroblasts, but none of the protocerebral (anterior brain) neuroblasts, display serial homology to neuroblasts in thoracic/abdominal neuromeres. Modifications in the molecular signature of serially homologous neuroblasts are likely to determine the segment-specific characteristics of their lineages. PMID:27095493

  7. The Drosophila anatomy ontology

    PubMed Central

    2013-01-01

    Background Anatomy ontologies are query-able classifications of anatomical structures. They provide a widely-used means for standardising the annotation of phenotypes and expression in both human-readable and programmatically accessible forms. They are also frequently used to group annotations in biologically meaningful ways. Accurate annotation requires clear textual definitions for terms, ideally accompanied by images. Accurate grouping and fruitful programmatic usage requires high-quality formal definitions that can be used to automate classification and check for errors. The Drosophila anatomy ontology (DAO) consists of over 8000 classes with broad coverage of Drosophila anatomy. It has been used extensively for annotation by a range of resources, but until recently it was poorly formalised and had few textual definitions. Results We have transformed the DAO into an ontology rich in formal and textual definitions in which the majority of classifications are automated and extensive error checking ensures quality. Here we present an overview of the content of the DAO, the patterns used in its formalisation, and the various uses it has been put to. Conclusions As a result of the work described here, the DAO provides a high-quality, queryable reference for the wild-type anatomy of Drosophila melanogaster and a set of terms to annotate data related to that anatomy. Extensive, well referenced textual definitions make it both a reliable and useful reference and ensure accurate use in annotation. Wide use of formal axioms allows a large proportion of classification to be automated and the use of consistency checking to eliminate errors. This increased formalisation has resulted in significant improvements to the completeness and accuracy of classification. The broad use of both formal and informal definitions make further development of the ontology sustainable and scalable. The patterns of formalisation used in the DAO are likely to be useful to developers of other

  8. Calcium and egg activation in Drosophila.

    PubMed

    Sartain, Caroline V; Wolfner, Mariana F

    2013-01-01

    In many animals, a rise in intracellular calcium levels is the trigger for egg activation, the process by which an arrested mature oocyte transitions to prepare for embryogenesis. In nearly all animals studied to date, this calcium rise, and thus egg activation, is triggered by the fertilizing sperm. However in the insects that have been examined, fertilization is not necessary to activate their oocytes. Rather, these insects' eggs activate as they transit through the female's reproductive tract, regardless of male contribution. Recent studies in Drosophila have shown that egg activation nevertheless requires calcium and that the downstream events and molecules of egg activation are also conserved, despite the difference in initial trigger. Genetic studies have uncovered essential roles for the calcium-dependent enzyme calcineurin and its regulator calcipressin, and have hinted at roles for calmodulin, in Drosophila egg activation. Physiological and in vitro studies have led to a model in which mechanical forces that impact the Drosophila oocyte as it moves through the reproductive tract triggers the influx of calcium from the external environment, thereby initiating egg activation. Future research will aim to test this model, as well as to determine the spatiotemporal dynamics of cytoplasmic calcium flux and mode of signal propagation in this unique system. PMID:23218670

  9. Imprinted control of gene activity in Drosophila.

    PubMed

    Golic, K G; Golic, M M; Pimpinelli, S

    1998-11-19

    Genetic imprinting is defined as a reversible, differential marking of genes or chromosomes that is determined by the sex of the parent from whom the genetic material is inherited [1]. Imprinting was first observed in insects where, in some species, most notably among the coccoids (scale insects and allies), the differential marking of paternally and maternally transmitted chromosome sets leads to inactivation or elimination of paternal chromosomes [2]. Imprinting is also widespread in plants and mammals [3,4], in which paternally and maternally inherited alleles may be differentially expressed. Despite imprinting having been discovered in insects, clear examples of parental imprinting are scarce in the model insect species Drosophila melanogaster. We describe a case of imprint-mediated control of gene expression in Drosophila. The imprinted gene - the white+ eye-color gene - is expressed at a low level when transmitted by males, and at a high level when transmitted by females. Thus, in common with coccoids, Drosophila is capable of generating an imprint, and can respond to that imprint by silencing the paternal allele. PMID:9822579

  10. Queuine metabolism and cadmium toxicity in Drosophila

    SciTech Connect

    Farkas, W.R.; Siard, T. ); Jacobson, K.B. )

    1991-03-11

    Queuine is a derivative of guanine found in the first position of the anticodon of the transfer RNAs for Asp, Asn, His and Tyr. The transcripts of these tRNAs contain a guanine in this position. This guanine is enzymatically excised and replaced by queuine. The ratio of queuine-containing or (q+) tRNA to its precursor or (q{minus}) tRNA changes throughout the Drosophila life cycle. in the egg 10% of the tRNA is (q+). During the three larval stages this ratio drops to zero. In the one day old adult it is about 10%. It has previously been shown that when flies are selected for the ability to grow in the presence of cadmium, the tolerant flies had 100% (q+) tRNA at the first day after pupation instead of 10%. However, it was not known whether the elevated level of (q+) tRNA was a coincidence or if the elevated levels of (q+) tRNA was protective. The authors explored this problem using germfree Drosophila. The first thing was to determine if Drosophila can synthesize queuine. Sterilized eggs were seeded onto sterile chemically defined medium. The flies were grown to the adult stage. This study showed that Drosophila like mammals cannot synthesize queuine. A second result of this research was the demonstration that the authors could alter the ratio of (q+) to (q{minus}) tRNA by adding exogenous queuine to the medium e.g. at 0.008 mM queuine the (q+) tRNA was 95% instead of {lt} 5% in the last instar stage. Finally, the authors investigated whether or not queuine gave protection against cadmium. The results were that when the flies were grown in the presence of 0.2 mM cadmium queuine at 0.008 mM gave a statistically significant increase in the number of survivors.

  11. Sexual circuitry in Drosophila.

    PubMed

    Auer, Thomas O; Benton, Richard

    2016-06-01

    The sexual behavior of Drosophila melanogaster is an outstanding paradigm to understand the molecular and neuronal basis of sophisticated animal actions. We discuss recent advances in our knowledge of the genetic hardwiring of the underlying neuronal circuitry, and how pertinent sensory cues are differentially detected and integrated in the male and female brain. We also consider how experience influences these circuits over short timescales, and the evolution of these pathways over longer timescales to endow species-specific sexual displays and responses. PMID:26851712

  12. Initial neurogenesis in Drosophila

    PubMed Central

    Hartenstein, Volker; Wodarz, Andreas

    2014-01-01

    Early neurogenesis comprises the phase of nervous system development during which neural progenitor cells are born. In early development, the embryonic ectoderm is subdivided by a conserved signaling mechanism into two main domains, the epidermal ectoderm and the neurectoderm. Subsequently, cells of the neurectoderm are internalized and form a cell layer of proliferating neural progenitors. In vertebrates, the entire neurectoderm folds into the embryo to give rise to the neural tube. In Drosophila and many other invertebrates, a subset of neurectodermal cells, called neuroblasts (NBs), delaminates and forms the neural primordium inside the embryo where they divide in an asymmetric, stem cell-like mode. The remainder of the neuroectodermal cells that stay behind at the surface loose their neurogenic potential and later give rise to the ventral part of the epidermis. The genetic and molecular analysis of the mechanisms controlling specification and proliferation of NBs in the Drosophila embryo, which played a significant part in pioneering the field of modern developmental neurobiology, represents the topic of this review. PMID:24014455

  13. Initial neurogenesis in Drosophila.

    PubMed

    Hartenstein, Volker; Wodarz, Andreas

    2013-01-01

    Early neurogenesis comprises the phase of nervous system development during which neural progenitor cells are born. In early development, the embryonic ectoderm is subdivided by a conserved signaling mechanism into two main domains, the epidermal ectoderm and the neurectoderm. Subsequently, cells of the neurectoderm are internalized and form a cell layer of proliferating neural progenitors. In vertebrates, the entire neurectoderm folds into the embryo to give rise to the neural tube. In Drosophila and many other invertebrates, a subset of neurectodermal cells, called neuroblasts (NBs), delaminates and forms the neural primordium inside the embryo where they divide in an asymmetric, stem cell-like mode. The remainder of the neurectodermal cells that stay behind at the surface loose their neurogenic potential and later give rise to the ventral part of the epidermis. The genetic and molecular analysis of the mechanisms controlling specification and proliferation of NBs in the Drosophila embryo, which played a significant part in pioneering the field of modern developmental neurobiology, represents the topic of this review. PMID:24014455

  14. Density dynamics of diverse Spiroplasma strains naturally infecting different species of Drosophila

    PubMed Central

    Haselkorn, Tamara S; Watts, Thomas D; Markow, Therese A

    2013-01-01

    Facultative heritable bacterial endosymbionts can have dramatic effects on their hosts, ranging from mutualistic to parasitic. Within-host bacterial endosymbiont density plays a critical role in maintenance of a symbiotic relationship, as it can affect levels of vertical transmission and expression of phenotypic effects, both of which influence the infection prevalence in host populations. Species of genus Drosophila are infected with Spiroplasma, whose characterized phenotypic effects range from that of a male-killing reproductive parasite to beneficial defensive endosymbiont. For many strains of Spiroplasma infecting at least 17 species of Drosophila, however, the phenotypic effects are obscure. The infection prevalence of these Spiroplasma vary within and among Drosophila species, and little is known about the within-host density dynamics of these diverse strains. To characterize the patterns of Spiroplasma density variation among Drosophila we used quantitative PCR to assess bacterial titer at various life stages of three species of Drosophila naturally-infected with two different types of Spiroplasma. For naturally infected Drosophila species we found that non-male-killing infections had consistently lower densities than the male-killing infection. The patterns of Spiroplasma titer change during aging varied among Drosophila species infected with different Spiroplasma strains. Bacterial density varied within and among populations of Drosophila, with individuals from the population with the highest prevalence of infection having the highest density. This density variation underscores the complex interaction of Spiroplasma strain and host genetic background in determining endosymbiont density. PMID:23846301

  15. Transmembrane segments of complement receptor 3 do not participate in cytotoxic activities but determine receptor structure required for action of Bordetella adenylate cyclase toxin.

    PubMed

    Wald, Tomas; Osickova, Adriana; Masin, Jiri; Liskova, Petra M; Petry-Podgorska, Inga; Matousek, Tomas; Sebo, Peter; Osicka, Radim

    2016-04-01

    Adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) of the whooping cough agent Bordetella pertussis penetrates phagocytes expressing the integrin complement receptor 3 (CR3, CD11b/CD18, α(M)β(2) or Mac-1). CyaA translocates its adenylate cyclase (AC) enzyme domain into cell cytosol and catalyzes unregulated conversion of ATP to cAMP, thereby subverting cellular signaling. In parallel, CyaA forms small cation-selective membrane pores that permeabilize cells for potassium efflux, contributing to cytotoxicity of CyaA and eventually provoking colloid-osmotic cell lysis. To investigate whether the single-pass α-helical transmembrane segments of CR3 subunits CD11b and CD18 do directly participate in AC domain translocation and/or pore formation by the toxin, we expressed in CHO cells variants of CR3 that contained artificial transmembrane segments, or lacked the transmembrane segment(s) at all. The results demonstrate that the transmembrane segments of CR3 are not directly involved in the cytotoxic activities of CyaA but serve for maintaining CR3 in a conformation that is required for efficient toxin binding and action. PMID:26802078

  16. Vibration damping for the Segmented Mirror Telescope

    NASA Astrophysics Data System (ADS)

    Maly, Joseph R.; Yingling, Adam J.; Griffin, Steven F.; Agrawal, Brij N.; Cobb, Richard G.; Chambers, Trevor S.

    2012-09-01

    The Segmented Mirror Telescope (SMT) at the Naval Postgraduate School (NPS) in Monterey is a next-generation deployable telescope, featuring a 3-meter 6-segment primary mirror and advanced wavefront sensing and correction capabilities. In its stowed configuration, the SMT primary mirror segments collapse into a small volume; once on location, these segments open to the full 3-meter diameter. The segments must be very accurately aligned after deployment and the segment surfaces are actively controlled using numerous small, embedded actuators. The SMT employs a passive damping system to complement the actuators and mitigate the effects of low-frequency (<40 Hz) vibration modes of the primary mirror segments. Each of the six segments has three or more modes in this bandwidth, and resonant vibration excited by acoustics or small disturbances on the structure can result in phase mismatches between adjacent segments thereby degrading image quality. The damping system consists of two tuned mass dampers (TMDs) for each of the mirror segments. An adjustable TMD with passive magnetic damping was selected to minimize sensitivity to changes in temperature; both frequency and damping characteristics can be tuned for optimal vibration mitigation. Modal testing was performed with a laser vibrometry system to characterize the SMT segments with and without the TMDs. Objectives of this test were to determine operating deflection shapes of the mirror and to quantify segment edge displacements; relative alignment of λ/4 or better was desired. The TMDs attenuated the vibration amplitudes by 80% and reduced adjacent segment phase mismatches to acceptable levels.

  17. Inertial properties of equine limb segments

    PubMed Central

    Nauwelaerts, Sandra; Allen, Whitney A; Lane, Jasmine M; Clayton, Hilary M

    2011-01-01

    Quantifying the dynamics of limb movements requires knowledge of the mass distribution between and within limb segments. We measured segment masses, positions of segmental center of mass and moments of inertia of the fore and hind limb segments for 38 horses of different breeds and sizes. After disarticulation by dissections, segments were weighed and the position of the center of mass was determined by suspension. Moment of inertia was measured using a trifilar pendulum. We found that mass distribution does not change with size for animals under 600 kg and report ratios of segmental masses to total body mass. For all segments, the scaling relationship between segmental mass and moment of inertia was predicted equally well or better by a 5/3 power fit than by the more classic mass multiplied by segmental length squared fit. Average values taken from previous studies generally confirmed our data but scaling relationships often needed to be revised. We did not detect an effect of morphotype on segment inertial properties. Differences in segmental inertial properties between published studies may depend more on segmental segmentation techniques than on size or body type of the horse. PMID:21355866

  18. Review: Thermal preference in Drosophila

    PubMed Central

    Dillon, Michael E.; Wang, George; Garrity, Paul A.; Huey, Raymond B.

    2009-01-01

    Environmental temperature strongly affects physiology of ectotherms. Small ectotherms, like Drosophila, cannot endogenously regulate body temperature so must rely on behavior to maintain body temperature within a physiologically permissive range. Here we review what is known about Drosophila thermal preference. Work on thermal behavior in this group is particularly exciting because it provides the opportunity to connect genes to neuromolecular mechanisms to behavior to fitness in the wild. PMID:20161211

  19. A kinetic analysis of Drosophila melanogaster dopa decarboxylase.

    PubMed

    Black, B C; Smarrelli, J

    1986-03-01

    The kinetic mechanism of dopa decarboxylase (3,4-dihydroxy-L-phenylalanine carboxy-lyase, EC 4.1.1.28) was investigated in Drosophila melanogaster. Based on initial velocity and product inhibition studies, an ordered reaction is proposed for dopa decarboxylase. This kinetic mechanism is interpreted in the context of measured enzyme activities and the catecholamine pools in Drosophila. The 1(2)amd gene is immediately adjacent to the gene coding for dopa decarboxylase (Ddc) and determines hypersensitivity to alpha-methyldopa in Drosophila. Dopa decarboxylase does not decarboxylate alpha-methyldopa and hence does not generate a toxic product capable of inhibiting 1(2)amd gene function. We propose that the 1(2)amd gene is involved with an unknown catecholamine pathway involving dopa but not dopamine. PMID:3081033

  20. A novel basic helix-loop-helix protein is expressed in muscle attachment sites of the Drosophila epidermis.

    PubMed Central

    Armand, P; Knapp, A C; Hirsch, A J; Wieschaus, E F; Cole, M D

    1994-01-01

    We have found that a novel basic helix-loop-helix (bHLH) protein is expressed almost exclusively in the epidermal attachments sites for the somatic muscles of Drosophila melanogaster. A Drosophila cDNA library was screened with radioactively labeled E12 protein, which can dimerize with many HLH proteins. One clone that emerged from this screen encoded a previously unknown protein of 360 amino acids, named delilah, that contains both basic and HLH domains, similar to a group of cellular transcription factors implicated in cell type determination. Delilah protein formed heterodimers with E12 that bind to the muscle creatine kinase promoter. In situ hybridization with the delilah cDNA localized the expression of the gene to a subset of cells in the epidermis which form a distinct pattern involving both the segmental boundaries and intrasegmental clusters. This pattern was coincident with the known sites of attachment of the somatic muscles to tendon cells in the epidermis. delilah expression persists in snail mutant embryos which lack mesoderm, indicating that expression of the gene was not induced by attachment of the underlying muscles. The similarity of this gene to other bHLH genes suggests that it plays an important role in the differentiation of epidermal cells into muscle attachment sites. Images PMID:8196652

  1. Segmental dynamics of the cytoplasmic domain of erythrocyte band 3 determined by time-resolved fluorescence anisotropy: sensitivity to pH and ligand binding.

    PubMed

    Thevenin, B J; Periasamy, N; Shohet, S B; Verkman, A S

    1994-03-01

    Interactions between the erythrocyte membrane and its skeleton are mediated primarily by binding of cytoskeletal components to a conformationally sensitive structure, the cytoplasmic domain of band 3 (cdb3). To examine the nanosecond segmental motions of cdb3, band 3 was labeled selectively by fluorescein maleimide at Cys-201 near the proposed hinge in cdb3 about which pH-dependent conformational changes occur. Time-resolved anisotropy of labeled cdb3 in isolated form and in stripped erythrocyte membranes was measured by parallel-acquisition frequency-domain microfluorimetry. Samples had a single-component fluorescein lifetime of approximately 4 ns. Multifrequency phase and modulation data (5-200 MHz) fitted well to a segmental motion model containing two correlation times (tau 1c and tau 2c) and two limiting anisotropies (r1infinity and r2infinity). Measurements in protease-cleaved and denatured samples indicated that tau 1c (100-150 ps) corresponded to rapid rotation of bound fluorescein and tau 2c (2-5 ns) corresponded to segmental motion of cdb3. Both motions were hindered as quantified by nonzero r1infinity and r2infinity. The strong pH dependence of segmental motion correlated with that of cdb3 conformation measured by intrinsic tryptophan fluorescence. Significant changes in cdb3 segmental motion occurred upon interactions with the small ligands 2,3-bisphosphoglycerate and calcium and several glycolytic enzymes known to bind to the N terminus of band 3. These time-resolved fluorescence measurements of the nanosecond segmental dynamics of a labeled membrane protein provide evidence for the sensitivity of cdb3 conformation to ligand binding and suggest long-range structural communication through cdb3. PMID:8127875

  2. Associations of Yeasts with Spotted-Wing Drosophila (Drosophila suzukii; Diptera: Drosophilidae) in Cherries and Raspberries

    PubMed Central

    Hernández, Alejandro; Zalom, Frank G.

    2012-01-01

    A rich history of investigation documents various Drosophila-yeast mutualisms, suggesting that Drosophila suzukii similarly has an association with a specific yeast species or community. To discover candidate yeast species, yeasts were isolated from larval frass, adult midguts, and fruit hosts of D. suzukii. Terminal restriction fragment length polymorphism (TRFLP) technology and decimal dilution plating were used to identify and determine the relative abundance of yeast species present in fruit juice samples that were either infested with D. suzukii or not infested. Yeasts were less abundant in uninfested than infested samples. A total of 126 independent yeast isolates were cultivated from frass, midguts, and fruit hosts of D. suzukii, representing 28 species of yeasts, with Hanseniaspora uvarum predominating. This suggests an association between D. suzukii and H. uvarum that could be utilized for pest management of the highly pestiferous D. suzukii. PMID:22582060

  3. Associations of yeasts with spotted-wing Drosophila (Drosophila suzukii; Diptera: Drosophilidae) in cherries and raspberries.

    PubMed

    Hamby, Kelly A; Hernández, Alejandro; Boundy-Mills, Kyria; Zalom, Frank G

    2012-07-01

    A rich history of investigation documents various Drosophila-yeast mutualisms, suggesting that Drosophila suzukii similarly has an association with a specific yeast species or community. To discover candidate yeast species, yeasts were isolated from larval frass, adult midguts, and fruit hosts of D. suzukii. Terminal restriction fragment length polymorphism (TRFLP) technology and decimal dilution plating were used to identify and determine the relative abundance of yeast species present in fruit juice samples that were either infested with D. suzukii or not infested. Yeasts were less abundant in uninfested than infested samples. A total of 126 independent yeast isolates were cultivated from frass, midguts, and fruit hosts of D. suzukii, representing 28 species of yeasts, with Hanseniaspora uvarum predominating. This suggests an association between D. suzukii and H. uvarum that could be utilized for pest management of the highly pestiferous D. suzukii. PMID:22582060

  4. Efficient graph-cut tattoo segmentation

    NASA Astrophysics Data System (ADS)

    Kim, Joonsoo; Parra, Albert; Li, He; Delp, Edward J.

    2015-03-01

    Law enforcement is interested in exploiting tattoos as an information source to identify, track and prevent gang-related crimes. Many tattoo image retrieval systems have been described. In a retrieval system tattoo segmentation is an important step for retrieval accuracy since segmentation removes background information in a tattoo image. Existing segmentation methods do not extract the tattoo very well when the background includes textures and color similar to skin tones. In this paper we describe a tattoo segmentation approach by determining skin pixels in regions near the tattoo. In these regions graph-cut segmentation using a skin color model and a visual saliency map is used to find skin pixels. After segmentation we determine which set of skin pixels are connected with each other that form a closed contour including a tattoo. The regions surrounded by the closed contours are considered tattoo regions. Our method segments tattoos well when the background includes textures and color similar to skin.

  5. P element excision in drosophila melanogaster and related drosophilids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The frequency of P element excision and the structure of the resulting excision products were determined in three drosophilid species, Drosophila melanogaster, D. virilis, and Chymomyza procnemis. A transient P element mobility assay was conducted in the cells of developing insect embryos, but unlik...

  6. Multiatlas segmentation as nonparametric regression.

    PubMed

    Awate, Suyash P; Whitaker, Ross T

    2014-09-01

    This paper proposes a novel theoretical framework to model and analyze the statistical characteristics of a wide range of segmentation methods that incorporate a database of label maps or atlases; such methods are termed as label fusion or multiatlas segmentation. We model these multiatlas segmentation problems as nonparametric regression problems in the high-dimensional space of image patches. We analyze the nonparametric estimator's convergence behavior that characterizes expected segmentation error as a function of the size of the multiatlas database. We show that this error has an analytic form involving several parameters that are fundamental to the specific segmentation problem (determined by the chosen anatomical structure, imaging modality, registration algorithm, and label-fusion algorithm). We describe how to estimate these parameters and show that several human anatomical structures exhibit the trends modeled analytically. We use these parameter estimates to optimize the regression estimator. We show that the expected error for large database sizes is well predicted by models learned on small databases. Thus, a few expert segmentations can help predict the database sizes required to keep the expected error below a specified tolerance level. Such cost-benefit analysis is crucial for deploying clinical multiatlas segmentation systems. PMID:24802528

  7. The developmental transcriptome of Drosophila melanogaster

    SciTech Connect

    University of Connecticut; Graveley, Brenton R.; Brooks, Angela N.; Carlson, Joseph W.; Duff, Michael O.; Landolin, Jane M.; Yang, Li; Artieri, Carlo G.; van Baren, Marijke J.; Boley, Nathan; Booth, Benjamin W.; Brown, James B.; Cherbas, Lucy; Davis, Carrie A.; Dobin, Alex; Li, Renhua; Lin, Wei; Malone, John H.; Mattiuzzo, Nicolas R.; Miller, David; Sturgill, David; Tuch, Brian B.; Zaleski, Chris; Zhang, Dayu; Blanchette, Marco; Dudoit, Sandrine; Eads, Brian; Green, Richard E.; Hammonds, Ann; Jiang, Lichun; Kapranov, Phil; Langton, Laura; Perrimon, Norbert; Sandler, Jeremy E.; Wan, Kenneth H.; Willingham, Aarron; Zhang, Yu; Zou, Yi; Andrews, Justen; Bicke, Peter J.; Brenner, Steven E.; Brent, Michael R.; Cherbas, Peter; Gingeras, Thomas R.; Hoskins, Roger A.; Kaufman, Thomas C.; Oliver, Brian; Celniker, Susan E.

    2010-12-02

    . Whereas, 20% of Drosophila genes are annotated as encoding alternatively spliced premRNAs, splice-junction microarray experiments indicate that this number is at least 40% (ref. 7). Determining the diversity of mRNAs generated by alternative promoters, alternative splicing and RNA editing will substantially increase the inferred protein repertoire. Non-coding RNA genes (ncRNAs) including short interfering RNAs (siRNAs) and microRNAS (miRNAs) (reviewed in ref. 10), and longer ncRNAs such as bxd (ref. 11) and rox (ref. 12), have important roles in gene regulation, whereas others such as small nucleolar RNAs (snoRNAs)and small nuclear RNAs (snRNAs) are important components of macromolecular machines such as the ribosome and spliceosome. The transcription and processing of these ncRNAs must also be fully documented and mapped. As part of the modENCODE project to annotate the functional elements of the D. melanogaster and Caenorhabditis elegans genomes, we used RNA-Seq and tiling microarrays to sample the Drosophila transcriptome at unprecedented depth throughout development from early embryo to ageing male and female adults. We report on a high-resolution view of the discovery, structure and dynamic expression of the D. melanogaster transcriptome.

  8. Effect of Exercise Intensity on Percent Body Fat Determined by Leg-to-Leg and Segmental Bioelectrical Impedance Analyses in Adults

    ERIC Educational Resources Information Center

    Andreacci, Joseph L.; Nagle, Trisha; Fitzgerald, Elise; Rawson, Eric S.; Dixon, Curt B.

    2013-01-01

    Purpose: We examined the impact that cycle ergometry exercise had on percent body fat (%BF) estimates when assessed using either leg-to-leg or segmental bioelectrical impedance analysis (LBIA; SBIA) and whether the intensity of the exercise bout impacts the %BF magnitude of change. Method: Seventy-four college-aged adults participated in this…

  9. On the determination of the effective transmission factor for stainless steel ovoid shielding segments and estimation of their shielding efficacy for the clinical situation.

    PubMed

    Verellen, D; De Neve, W; Van den Heuvel, F; Storme, G; Coen, V; Coghe, M

    1994-11-01

    Commercially available ovoid tubes for gynecological applications used in conjunction with the microSelectron-HDR (Nucletron International B.V., Waardegelder 1, 3905 TH Veenendaal, The Netherlands) for 192Ir sources, allow for shielding. Publications concerning the transmission properties of these 4.5-mm thick stainless steel (AISI number 303/304) shielding segments are scarce and not compatible for implementation in treatment planning. Therefore the effect of shielding on dose distribution is unknown. The effective transmission factor has been measured and implemented in the planning computations. Screening efficacy was evaluated on 20 actual treatment plans, analyzing dose reduction to critical tissue and comparing dose distribution in planes relevant for this particular application. Due to high transmission (effective transmission factor = 0.85), stainless steel screening segments only provide low, local dose reductions of maximum 15%. A new approach with regard to optimization and source configuration is needed to reduce dose to vulnerable tissue, exploiting the screening segments to a maximum extent. Better shielding, especially at the midline (plane bisecting the ovoids) could be expected by using shielding segments with other geometrical characteristics. PMID:7891627

  10. Mapping Linked Genes in "Drosophila Melanogaster" Using Data from the F2 Generation of a Dihybrid Cross

    ERIC Educational Resources Information Center

    Marshall, Pamela A.

    2008-01-01

    "Drosophila melanogaster" is a commonly utilized organism for testing hypotheses about inheritance of traits. Students in both high school and university labs study the genetics of inheritance by analyzing offspring of appropriate "Drosophila" crosses to determine inheritance patterns, including gene linkage. However, most genetics investigations…

  11. The open for business model of the bithorax complex in Drosophila.

    PubMed

    Maeda, Robert K; Karch, François

    2015-09-01

    After nearly 30 years of effort, Ed Lewis published his 1978 landmark paper in which he described the analysis of a series of mutations that affect the identity of the segments that form along the anterior-posterior (AP) axis of the fly (Lewis 1978). The mutations behaved in a non-canonical fashion in complementation tests, forming what Ed Lewis called a "pseudo-allelic" series. Because of this, he never thought that the mutations represented segment-specific genes. As all of these mutations were grouped to a particular area of the Drosophila third chromosome, the locus became known of as the bithorax complex (BX-C). One of the key findings of Lewis' article was that it revealed for the first time, to a wide scientific audience, that there was a remarkable correlation between the order of the segment-specific mutations along the chromosome and the order of the segments they affected along the AP axis. In Ed Lewis' eyes, the mutants he discovered affected "segment-specific functions" that were sequentially activated along the chromosome as one moves from anterior to posterior along the body axis (the colinearity concept now cited in elementary biology textbooks). The nature of the "segment-specific functions" started to become clear when the BX-C was cloned through the pioneering chromosomal walk initiated in the mid 1980s by the Hogness and Bender laboratories (Bender et al. 1983a; Karch et al. 1985). Through this molecular biology effort, and along with genetic characterizations performed by Gines Morata's group in Madrid (Sanchez-Herrero et al. 1985) and Robert Whittle's in Sussex (Tiong et al. 1985), it soon became clear that the whole BX-C encoded only three protein-coding genes (Ubx, abd-A, and Abd-B). Later, immunostaining against the Ubx protein hinted that the segment-specific functions could, in fact, be cis-regulatory elements regulating the expression of the three protein-coding genes. In 1987, Peifer, Karch, and Bender proposed a comprehensive model of

  12. Estimating divergence dates and substitution rates in the Drosophila phylogeny.

    PubMed

    Obbard, Darren J; Maclennan, John; Kim, Kang-Wook; Rambaut, Andrew; O'Grady, Patrick M; Jiggins, Francis M

    2012-11-01

    An absolute timescale for evolution is essential if we are to associate evolutionary phenomena, such as adaptation or speciation, with potential causes, such as geological activity or climatic change. Timescales in most phylogenetic studies use geologically dated fossils or phylogeographic events as calibration points, but more recently, it has also become possible to use experimentally derived estimates of the mutation rate as a proxy for substitution rates. The large radiation of drosophilid taxa endemic to the Hawaiian islands has provided multiple calibration points for the Drosophila phylogeny, thanks to the "conveyor belt" process by which this archipelago forms and is colonized by species. However, published date estimates for key nodes in the Drosophila phylogeny vary widely, and many are based on simplistic models of colonization and coalescence or on estimates of island age that are not current. In this study, we use new sequence data from seven species of Hawaiian Drosophila to examine a range of explicit coalescent models and estimate substitution rates. We use these rates, along with a published experimentally determined mutation rate, to date key events in drosophilid evolution. Surprisingly, our estimate for the date for the most recent common ancestor of the genus Drosophila based on mutation rate (25-40 Ma) is closer to being compatible with independent fossil-derived dates (20-50 Ma) than are most of the Hawaiian-calibration models and also has smaller uncertainty. We find that Hawaiian-calibrated dates are extremely sensitive to model choice and give rise to point estimates that range between 26 and 192 Ma, depending on the details of the model. Potential problems with the Hawaiian calibration may arise from systematic variation in the molecular clock due to the long generation time of Hawaiian Drosophila compared with other Drosophila and/or uncertainty in linking island formation dates with colonization dates. As either source of error will

  13. Estimating Divergence Dates and Substitution Rates in the Drosophila Phylogeny

    PubMed Central

    Obbard, Darren J.; Maclennan, John; Kim, Kang-Wook; Rambaut, Andrew; O’Grady, Patrick M.; Jiggins, Francis M.

    2012-01-01

    An absolute timescale for evolution is essential if we are to associate evolutionary phenomena, such as adaptation or speciation, with potential causes, such as geological activity or climatic change. Timescales in most phylogenetic studies use geologically dated fossils or phylogeographic events as calibration points, but more recently, it has also become possible to use experimentally derived estimates of the mutation rate as a proxy for substitution rates. The large radiation of drosophilid taxa endemic to the Hawaiian islands has provided multiple calibration points for the Drosophila phylogeny, thanks to the "conveyor belt" process by which this archipelago forms and is colonized by species. However, published date estimates for key nodes in the Drosophila phylogeny vary widely, and many are based on simplistic models of colonization and coalescence or on estimates of island age that are not current. In this study, we use new sequence data from seven species of Hawaiian Drosophila to examine a range of explicit coalescent models and estimate substitution rates. We use these rates, along with a published experimentally determined mutation rate, to date key events in drosophilid evolution. Surprisingly, our estimate for the date for the most recent common ancestor of the genus Drosophila based on mutation rate (25–40 Ma) is closer to being compatible with independent fossil-derived dates (20–50 Ma) than are most of the Hawaiian-calibration models and also has smaller uncertainty. We find that Hawaiian-calibrated dates are extremely sensitive to model choice and give rise to point estimates that range between 26 and 192 Ma, depending on the details of the model. Potential problems with the Hawaiian calibration may arise from systematic variation in the molecular clock due to the long generation time of Hawaiian Drosophila compared with other Drosophila and/or uncertainty in linking island formation dates with colonization dates. As either source of error will

  14. Assaying Blood Cell Populations of the Drosophila melanogaster Larva

    PubMed Central

    Petraki, Sophia; Alexander, Brandy; Brückner, Katja

    2015-01-01

    In vertebrates, hematopoiesis is regulated by inductive microenvironments (niches). Likewise, in the invertebrate model organism Drosophila melanogaster, inductive microenvironments known as larval Hematopoietic Pockets (HPs) have been identified as anatomical sites for the development and regulation of blood cells (hemocytes), in particular of the self-renewing macrophage lineage. HPs are segmentally repeated pockets between the epidermis and muscle layers of the larva, which also comprise sensory neurons of the peripheral nervous system. In the larva, resident (sessile) hemocytes are exposed to anti-apoptotic, adhesive and proliferative cues from these sensory neurons and potentially other components of the HPs, such as the lining muscle and epithelial layers. During normal development, gradual release of resident hemocytes from the HPs fuels the population of circulating hemocytes, which culminates in the release of most of the resident hemocytes at the beginning of metamorphosis. Immune assaults, physical injury or mechanical disturbance trigger the premature release of resident hemocytes into circulation. The switch of larval hemocytes between resident locations and circulation raises the need for a common standard/procedure to selectively isolate and quantify these two populations of blood cells from single Drosophila larvae. Accordingly, this protocol describes an automated method to release and quantify the resident and circulating hemocytes from single larvae. The method facilitates ex vivo approaches, and may be adapted to serve a variety of developmental stages of Drosophila and other invertebrate organisms. PMID:26650404

  15. The morphogen Decapentaplegic employs a two-tier mechanism to activate target retinal determining genes during ectopic eye formation in Drosophila.

    PubMed

    Aggarwal, Poonam; Gera, Jayati; Mandal, Lolitika; Mandal, Sudip

    2016-01-01

    Understanding the role of morphogen in activating its target genes, otherwise epigenetically repressed, during change in cell fate specification is a very fascinating yet relatively unexplored domain. Our in vivo loss-of-function genetic analyses reveal that specifically during ectopic eye formation, the morphogen Decapentaplegic (Dpp), in conjunction with the canonical signaling responsible for transcriptional activation of retinal determining (RD) genes, triggers another signaling cascade. Involving dTak1 and JNK, this pathway down-regulates the expression of polycomb group of genes to do away with their repressive role on RD genes. Upon genetic inactivation of members of this newly identified pathway, the canonical Dpp signaling fails to trigger RD gene expression beyond a threshold, critical for ectopic photoreceptor differentiation. Moreover, the drop in ectopic RD gene expression and subsequent reduction in ectopic photoreceptor differentiation resulting from inactivation of dTak1 can be rescued by down-regulating the expression of polycomb group of genes. Our results unravel an otherwise unknown role of morphogen in coordinating simultaneous transcriptional activation and de-repression of target genes implicating its importance in cellular plasticity. PMID:27270790

  16. The morphogen Decapentaplegic employs a two-tier mechanism to activate target retinal determining genes during ectopic eye formation in Drosophila

    PubMed Central

    Aggarwal, Poonam; Gera, Jayati; Mandal, Lolitika; Mandal, Sudip

    2016-01-01

    Understanding the role of morphogen in activating its target genes, otherwise epigenetically repressed, during change in cell fate specification is a very fascinating yet relatively unexplored domain. Our in vivo loss-of-function genetic analyses reveal that specifically during ectopic eye formation, the morphogen Decapentaplegic (Dpp), in conjunction with the canonical signaling responsible for transcriptional activation of retinal determining (RD) genes, triggers another signaling cascade. Involving dTak1 and JNK, this pathway down-regulates the expression of polycomb group of genes to do away with their repressive role on RD genes. Upon genetic inactivation of members of this newly identified pathway, the canonical Dpp signaling fails to trigger RD gene expression beyond a threshold, critical for ectopic photoreceptor differentiation. Moreover, the drop in ectopic RD gene expression and subsequent reduction in ectopic photoreceptor differentiation resulting from inactivation of dTak1 can be rescued by down-regulating the expression of polycomb group of genes. Our results unravel an otherwise unknown role of morphogen in coordinating simultaneous transcriptional activation and de-repression of target genes implicating its importance in cellular plasticity. PMID:27270790

  17. A model for shape generation by strain and cell-cell adhesion in the epithelium of an arthropod leg segment.

    PubMed

    Mittenthal, J E; Mazo, R M

    1983-02-01

    We present a model for the energetic factors determining the most stable shape of a tubular epithelium such as the hypodermis of an arthropod leg segment. The model uses the analysis by Steinberg (1963) of rearrangement of cells in aggregates under the influence of differential adhesion, combining this analysis with the assumption that the epithelium behaves as an elastic sheet. The epithelium is assumed to consist of blocks of cells with different adhesive affinities, which remain unmixed in a quilt pattern. Rearrangement of cells within each block can adjust the shape of the tube by changing the shapes of the blocks. By means of such rearrangements the tube develops that shape which minimizes a free energy. The free energy is the difference between the energy of mechanical strain due to bending of the epithelium and the work of adhesion among cells. Minimization of the free energy for a cylindrical segment yields a scaling relation involving the length and radius of the segment. Leg segments of Drosophila conformed approximately to this relation, with deviations which suggest that a whole-limb pattern of adhesive affinities modulates the shaping effects of an adhesive pattern repeated in each leg segment. The model also predicts a transient deformation in an epithelium following a grafting operation. For example, deleting a slab of tissue from a tubular segment and reuniting the cut ends should produce a constriction of the tube at the host-graft junction. We propose that patterns of strain and adhesion can provide positional information which regulates subsequent development. Local increases in strain or adhesive disparity may stimulate mitoses; the resulting changes in distribution of cells will affect morphogenesis. PMID:6834865

  18. Pre-analytical and analytical variation of drug determination in segmented hair using ultra-performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Nielsen, Marie Katrine Klose; Johansen, Sys Stybe; Linnet, Kristian

    2014-01-01

    Assessment of total uncertainty of analytical methods for the measurements of drugs in human hair has mainly been derived from the analytical variation. However, in hair analysis several other sources of uncertainty will contribute to the total uncertainty. Particularly, in segmental hair analysis pre-analytical variations associated with the sampling and segmentation may be significant factors in the assessment of the total uncertainty budget. The aim of this study was to develop and validate a method for the analysis of 31 common drugs in hair using ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) with focus on the assessment of both the analytical and pre-analytical sampling variations. The validated method was specific, accurate (80-120%), and precise (CV≤20%) across a wide linear concentration range from 0.025-25 ng/mg for most compounds. The analytical variation was estimated to be less than 15% for almost all compounds. The method was successfully applied to 25 segmented hair specimens from deceased drug addicts showing a broad pattern of poly-drug use. The pre-analytical sampling variation was estimated from the genuine duplicate measurements of two bundles of hair collected from each subject after subtraction of the analytical component. For the most frequently detected analytes, the pre-analytical variation was estimated to be 26-69%. Thus, the pre-analytical variation was 3-7 folds larger than the analytical variation (7-13%) and hence the dominant component in the total variation (29-70%). The present study demonstrated the importance of including the pre-analytical variation in the assessment of the total uncertainty budget and in the setting of the 95%-uncertainty interval (±2CVT). Excluding the pre-analytical sampling variation could significantly affect the interpretation of results from segmental hair analysis. PMID:24378297

  19. A dynamic deep sleep stage in Drosophila.

    PubMed

    van Alphen, Bart; Yap, Melvyn H W; Kirszenblat, Leonie; Kottler, Benjamin; van Swinderen, Bruno

    2013-04-17

    How might one determine whether simple animals such as flies sleep in stages? Sleep in mammals is a dynamic process involving different stages of sleep intensity, and these are typically associated with measurable changes in brain activity (Blake and Gerard, 1937; Rechtschaffen and Kales, 1968; Webb and Agnew, 1971). Evidence for different sleep stages in invertebrates remains elusive, even though it has been well established that many invertebrate species require sleep (Campbell and Tobler, 1984; Hendricks et al., 2000; Shaw et al., 2000; Sauer et al., 2003). Here we used electrophysiology and arousal-testing paradigms to show that the fruit fly, Drosophila melanogaster, transitions between deeper and lighter sleep within extended bouts of inactivity, with deeper sleep intensities after ∼15 and ∼30 min of inactivity. As in mammals, the timing and intensity of these dynamic sleep processes in flies is homeostatically regulated and modulated by behavioral experience. Two molecules linked to synaptic plasticity regulate the intensity of the first deep sleep stage. Optogenetic upregulation of cyclic adenosine monophosphate during the day increases sleep intensity at night, whereas loss of function of a molecule involved in synaptic pruning, the fragile-X mental retardation protein, increases sleep intensity during the day. Our results show that sleep is not homogenous in insects, and suggest that waking behavior and the associated synaptic plasticity mechanisms determine the timing and intensity of deep sleep stages in Drosophila. PMID:23595750

  20. Why Drosophila to Study Phototransduction?

    PubMed Central

    Pak, William L.

    2010-01-01

    This review recounts the early history of Drosophila phototransduction genetics, covering the period between approximately 1966 to 1979. Early in this period, the author felt that there was an urgent need for a new approach in phototransduction research. Through inputs from a number of colleagues, he was led to consider isolating Drosophila mutants that are defective in the electroretinogram. Thanks to the efforts of dedicated associates and technical staff, by the end of this period, he was able to accumulate a large number of such mutants. Particularly important in this effort was the use of the mutant assay protocol based on the “prolonged depolarizing afterpotential.” This collection of mutants formed the basis of the subsequent intensive investigations of the Drosophila phototransduction cascade by many investigators. PMID:20536286

  1. Micromechanics of Drosophila Audition

    NASA Astrophysics Data System (ADS)

    Göpfert, M. C.; Robert, D.

    2003-02-01

    An analysis is presented of the auditory micromechanics of the fruit fly Drosophila melanogaster. In this animal, the distal part of the antenna constitutes a resonantly tuned sound receiver, the vibrations of which are transduced by a chordotonal sense organ in the antenna's base. Analyzing the mechanical behavior of the antennal receiver by means of microscanning laser Doppler vibrometry, we show that the auditory system of wild-type flies exhibits a hardening stiffness nonlinearity and spontaneously generates oscillations in the absence of external stimuli. According to the deprivation of these mechanical properties in mechanosensory mutants, the receiver's nonlinearity and oscillation activity are introduced by chordotonal auditory neurons. Requiring the mechanoreceptor-specific extracellular linker protein No-mechanoreceptor-potential-A (NompA), NompC mechanosensory transduction channels, Beethoven (Btv), and Touch-insensitive-larva-B (TilB), nonlinearity and oscillation activity of the fly's antennal receiver depend on prominent components of the auditory transduction machinery and seem to originate from motility of auditory receptor cilia.

  2. Retinal differentiation in Drosophila.

    PubMed

    Treisman, Jessica E

    2013-07-01

    Drosophila eye development has been extensively studied, due to the ease of genetic screens for mutations disrupting this process. The eye imaginal disc is specified during embryonic and larval development by the Pax6 homolog Eyeless and a network of downstream transcription factors. Expression of these factors is regulated by signaling molecules and also indirectly by growth of the eye disc. Differentiation of photoreceptor clusters initiates in the third larval instar at the posterior of the eye disc and progresses anteriorly, driven by the secreted protein Hedgehog. Within each cluster, the combined activities of Hedgehog signaling and Notch-mediated lateral inhibition induce and refine the expression of the transcription factor Atonal, which specifies the founding R8 photoreceptor of each ommatidium. Seven additional photoreceptors, followed by cone and pigment cells, are successively recruited by the signaling molecules Spitz, Delta, and Bride of sevenless. Combinations of these signals and of intrinsic transcription factors give each ommatidial cell its specific identity. During the pupal stages, rhodopsins are expressed, and the photoreceptors and accessory cells take on their final positions and morphologies to form the adult retina. Over the past few decades, the genetic analysis of this small number of cell types arranged in a repetitive structure has allowed a remarkably detailed understanding of the basic mechanisms controlling cell differentiation and morphological rearrangement. PMID:24014422

  3. Genetic basis of sex-specific color pattern variation in Drosophila malerkotliana.

    PubMed

    Ng, Chen Siang; Hamilton, Andrew M; Frank, Amanda; Barmina, Olga; Kopp, Artyom

    2008-09-01

    Pigmentation is a rapidly evolving trait that can play important roles in mimicry, sexual selection, thermoregulation, and other adaptive processes in many groups of animals. In Drosophila, pigmentation can differ dramatically among closely related taxa, presenting a good opportunity to dissect the genetic changes underlying species divergence. In this report, we investigate the genetic basis of color pattern variation between two allopatric subspecies of Drosophila malerkotliana, a widespread member of the ananassae species subgroup. In D. malerkotliana malerkotliana, the last three abdominal segments are darkly pigmented in males but not in females, while in D. malerkotliana pallens both sexes lack dark pigmentation. Composite interval mapping in F2 hybrid progeny shows that this difference is largely controlled by three quantitative trait loci (QTL) located on the 2L chromosome arm, which is homologous to the 3R of D. melanogaster (Muller element E). Using highly recombinant introgression strains produced by repeated backcrossing and phenotypic selection, we show that these QTL do not correspond to any of the candidate genes known to be involved in pigment patterning and synthesis in Drosophila. These results, in combination with similar analyses in other Drosophila species, indicate that different genetic and molecular changes are responsible for the evolution of similar phenotypic traits in different lineages. This feature makes Drosophila color patterns a powerful model for investigating how the genetic basis of trait evolution is influenced by the intrinsic organization of regulatory pathways controlling the development of these traits. PMID:18723880

  4. RSRM Segment Train Derailment and Recovery

    NASA Technical Reports Server (NTRS)

    Taylor Jr., Robert H.; McConnaugghey, Paul K.; Beaman, David E.; Moore, Dennis R.; Reed, Harry

    2008-01-01

    On May 2, 2007, a freight train carrying segments of the space shuttle's solid rocket boosters derailed in Myrtlewood, Alabama, after a rail trestle collapsed. The train was carrying Reusable Solid Rocket Motors (RSRM) 98 center and forward segments (STS-120) and RSRM 99 aft segments (STS-122). Initially, it was not known if the segments had been seriously damaged. Four segments dropped approximately 10 feet when the trestle collapsed and one of those four rolled off the track onto its side. The exit cones and the other four segments, not yet on the trestle, remained on solid ground. ATK and NASA immediately dispatched an investigation and recovery team to determine the safety of the situation and eventually the usability of the segments and exit cones for flight. Instrumentation on each segment provided invaluable data to determine the acceleration loads imparted into each loaded segment and exit cone. This paper details the incident, recovery plan and the team work that created a success story that ended with the safe launch of STS120 using the four center segments and the launch of STS122 using the Aft exit cones assemblies.

  5. Segment alignment control system

    NASA Technical Reports Server (NTRS)

    Aubrun, JEAN-N.; Lorell, Ken R.

    1988-01-01

    The segmented primary mirror for the LDR will require a special segment alignment control system to precisely control the orientation of each of the segments so that the resulting composite reflector behaves like a monolith. The W.M. Keck Ten Meter Telescope will utilize a primary mirror made up of 36 actively controlled segments. Thus the primary mirror and its segment alignment control system are directly analogous to the LDR. The problems of controlling the segments in the face of disturbances and control/structures interaction, as analyzed for the TMT, are virtually identical to those for the LDR. The two systems are briefly compared.

  6. Genetic and Environmental Control of Neurodevelopmental Robustness in Drosophila

    PubMed Central

    Mellert, David J.; Williamson, W. Ryan; Shirangi, Troy R.; Card, Gwyneth M.; Truman, James W.

    2016-01-01

    Interindividual differences in neuronal wiring may contribute to behavioral individuality and affect susceptibility to neurological disorders. To investigate the causes and potential consequences of wiring variation in Drosophila melanogaster, we focused on a hemilineage of ventral nerve cord interneurons that exhibits morphological variability. We find that late-born subclasses of the 12A hemilineage are highly sensitive to genetic and environmental variation. Neurons in the second thoracic segment are particularly variable with regard to two developmental decisions, whereas its segmental homologs are more robust. This variability “hotspot” depends on Ultrabithorax expression in the 12A neurons, indicating variability is cell-intrinsic and under genetic control. 12A development is more variable and sensitive to temperature in long-established laboratory strains than in strains recently derived from the wild. Strains with a high frequency of one of the 12A variants also showed a high frequency of animals with delayed spontaneous flight initiation, whereas other wing-related behaviors did not show such a correlation and were thus not overtly affected by 12A variation. These results show that neurodevelopmental robustness is variable and under genetic control in Drosophila and suggest that the fly may serve as a model for identifying conserved gene pathways that stabilize wiring in stressful developmental environments. Moreover, some neuronal lineages are variation hotspots and thus may be more amenable to evolutionary change. PMID:27223118

  7. Optimal segmentation and packaging process

    DOEpatents

    Kostelnik, K.M.; Meservey, R.H.; Landon, M.D.

    1999-08-10

    A process for improving packaging efficiency uses three dimensional, computer simulated models with various optimization algorithms to determine the optimal segmentation process and packaging configurations based on constraints including container limitations. The present invention is applied to a process for decontaminating, decommissioning (D and D), and remediating a nuclear facility involving the segmentation and packaging of contaminated items in waste containers in order to minimize the number of cuts, maximize packaging density, and reduce worker radiation exposure. A three-dimensional, computer simulated, facility model of the contaminated items are created. The contaminated items are differentiated. The optimal location, orientation and sequence of the segmentation and packaging of the contaminated items is determined using the simulated model, the algorithms, and various constraints including container limitations. The cut locations and orientations are transposed to the simulated model. The contaminated items are actually segmented and packaged. The segmentation and packaging may be simulated beforehand. In addition, the contaminated items may be cataloged and recorded. 3 figs.

  8. Proprioceptive feedback determines visuomotor gain in Drosophila.

    PubMed

    Bartussek, Jan; Lehmann, Fritz-Olaf

    2016-01-01

    Multisensory integration is a prerequisite for effective locomotor control in most animals. Especially, the impressive aerial performance of insects relies on rapid and precise integration of multiple sensory modalities that provide feedback on different time scales. In flies, continuous visual signalling from the compound eyes is fused with phasic proprioceptive feedback to ensure precise neural activation of wing steering muscles (WSM) within narrow temporal phase bands of the stroke cycle. This phase-locked activation relies on mechanoreceptors distributed over wings and gyroscopic halteres. Here we investigate visual steering performance of tethered flying fruit flies with reduced haltere and wing feedback signalling. Using a flight simulator, we evaluated visual object fixation behaviour, optomotor altitude control and saccadic escape reflexes. The behavioural assays show an antagonistic effect of wing and haltere signalling on visuomotor gain during flight. Compared with controls, suppression of haltere feedback attenuates while suppression of wing feedback enhances the animal's wing steering range. Our results suggest that the generation of motor commands owing to visual perception is dynamically controlled by proprioception. We outline a potential physiological mechanism based on the biomechanical properties of WSM and sensory integration processes at the level of motoneurons. Collectively, the findings contribute to our general understanding how moving animals integrate sensory information with dynamically changing temporal structure. PMID:26909184

  9. Proprioceptive feedback determines visuomotor gain in Drosophila

    PubMed Central

    Bartussek, Jan; Lehmann, Fritz-Olaf

    2016-01-01

    Multisensory integration is a prerequisite for effective locomotor control in most animals. Especially, the impressive aerial performance of insects relies on rapid and precise integration of multiple sensory modalities that provide feedback on different time scales. In flies, continuous visual signalling from the compound eyes is fused with phasic proprioceptive feedback to ensure precise neural activation of wing steering muscles (WSM) within narrow temporal phase bands of the stroke cycle. This phase-locked activation relies on mechanoreceptors distributed over wings and gyroscopic halteres. Here we investigate visual steering performance of tethered flying fruit flies with reduced haltere and wing feedback signalling. Using a flight simulator, we evaluated visual object fixation behaviour, optomotor altitude control and saccadic escape reflexes. The behavioural assays show an antagonistic effect of wing and haltere signalling on visuomotor gain during flight. Compared with controls, suppression of haltere feedback attenuates while suppression of wing feedback enhances the animal’s wing steering range. Our results suggest that the generation of motor commands owing to visual perception is dynamically controlled by proprioception. We outline a potential physiological mechanism based on the biomechanical properties of WSM and sensory integration processes at the level of motoneurons. Collectively, the findings contribute to our general understanding how moving animals integrate sensory information with dynamically changing temporal structure. PMID:26909184

  10. Fluctuation-Driven Neural Dynamics Reproduce Drosophila Locomotor Patterns

    PubMed Central

    Cruchet, Steeve; Gustafson, Kyle; Benton, Richard; Floreano, Dario

    2015-01-01

    The neural mechanisms determining the timing of even simple actions, such as when to walk or rest, are largely mysterious. One intriguing, but untested, hypothesis posits a role for ongoing activity fluctuations in neurons of central action selection circuits that drive animal behavior from moment to moment. To examine how fluctuating activity can contribute to action timing, we paired high-resolution measurements of freely walking Drosophila melanogaster with data-driven neural network modeling and dynamical systems analysis. We generated fluctuation-driven network models whose outputs—locomotor bouts—matched those measured from sensory-deprived Drosophila. From these models, we identified those that could also reproduce a second, unrelated dataset: the complex time-course of odor-evoked walking for genetically diverse Drosophila strains. Dynamical models that best reproduced both Drosophila basal and odor-evoked locomotor patterns exhibited specific characteristics. First, ongoing fluctuations were required. In a stochastic resonance-like manner, these fluctuations allowed neural activity to escape stable equilibria and to exceed a threshold for locomotion. Second, odor-induced shifts of equilibria in these models caused a depression in locomotor frequency following olfactory stimulation. Our models predict that activity fluctuations in action selection circuits cause behavioral output to more closely match sensory drive and may therefore enhance navigation in complex sensory environments. Together these data reveal how simple neural dynamics, when coupled with activity fluctuations, can give rise to complex patterns of animal behavior. PMID:26600381

  11. Analysis of murine HOXA-2 activity in Drosophila melanogaster.

    PubMed

    Percival-Smith, A; Bondy, J A

    1999-01-01

    The murine HOXA-2 protein shares amino acid sequence similarity with Drosophila Proboscipedia (PB). In this paper, we test whether HOXA-2 and PB are functionally equivalent in Drosophila. In Drosophila, PB inhibits SCR activity required for larval T1 beard formation and adult tarsus formation and is required for maxillary palp and proboscis formation. HOXA-2 expressed from a heat-shock promoter weakly suppressed SCR activity required for T1 beard formation. But interestingly neither PB nor HOXA-2 expressed from a heat-shock promoter suppressed murine HOXA-5 activity, the murine SCR homologue, from inducing ectopic T1 beards in T2 and T3, indicating that HOXA-5 does not interact with PB. HOXA-2 activity expressed from the Tubulin alpha 1 promoter modified the pb null phenotype resulting in a proboscis-to-arista transformation, indicating that HOXA-2 was able to suppress SCR activity required for tarsus formation. However, HOXA-2 expressed from a Tubulin alpha 1 promoter was unable to direct maxillary palp determination when either ectopically expressed in the antenna or in the maxillary palp primordia of a pb null mutant. HOXA-2 was also unable to rescue pseudotrachea formation in a pb null mutant. These results indicate that the only activity that PB and HOXA-2 weakly share is the inhibition of SCR activity, and that murine HOXA-5 and Drosophila SCR do not share inhibition by PB activity. PMID:10322642

  12. Gr33a modulates Drosophila male courtship preference.

    PubMed

    Hu, Yujia; Han, Yi; Shao, Yingyao; Wang, Xingjun; Ma, Yeqing; Ling, Erjun; Xue, Lei

    2015-01-01

    In any gamogenetic species, attraction between individuals of the opposite sex promotes reproductive success that guarantees their thriving. Consequently, mate determination between two sexes is effortless for an animal. However, choosing a spouse from numerous attractive partners of the opposite sex needs deliberation. In Drosophila melanogaster, both younger virgin females and older ones are equally liked options to males; nevertheless, when given options, males prefer younger females to older ones. Non-volatile cuticular hydrocarbons, considered as major pheromones in Drosophila, constitute females' sexual attraction that act through males' gustatory receptors (Grs) to elicit male courtship. To date, only a few putative Grs are known to play roles in male courtship. Here we report that loss of Gr33a function or abrogating the activity of Gr33a neurons does not disrupt male-female courtship, but eliminates males' preference for younger mates. Furthermore, ectopic expression of human amyloid precursor protein (APP) in Gr33a neurons abolishes males' preference behavior. Such function of APP is mediated by the transcription factor forkhead box O (dFoxO). These results not only provide mechanistic insights into Drosophila male courtship preference, but also establish a novel Drosophila model for Alzheimer's disease (AD). PMID:25586066

  13. Gr33a Modulates Drosophila Male Courtship Preference

    PubMed Central

    Hu, Yujia; Han, Yi; Shao, Yingyao; Wang, Xingjun; Ma, Yeqing; Ling, Erjun; Xue, Lei

    2015-01-01

    In any gamogenetic species, attraction between individuals of the opposite sex promotes reproductive success that guarantees their thriving. Consequently, mate determination between two sexes is effortless for an animal. However, choosing a spouse from numerous attractive partners of the opposite sex needs deliberation. In Drosophila melanogaster, both younger virgin females and older ones are equally liked options to males; nevertheless, when given options, males prefer younger females to older ones. Non-volatile cuticular hydrocarbons, considered as major pheromones in Drosophila, constitute females' sexual attraction that act through males' gustatory receptors (Grs) to elicit male courtship. To date, only a few putative Grs are known to play roles in male courtship. Here we report that loss of Gr33a function or abrogating the activity of Gr33a neurons does not disrupt male-female courtship, but eliminates males' preference for younger mates. Furthermore, ectopic expression of human amyloid precursor protein (APP) in Gr33a neurons abolishes males' preference behavior. Such function of APP is mediated by the transcription factor forkhead box O (dFoxO). These results not only provide mechanistic insights into Drosophila male courtship preference, but also establish a novel Drosophila model for Alzheimer's disease (AD). PMID:25586066

  14. Substrate-borne vibratory communication during courtship in Drosophila melanogaster.

    PubMed

    Fabre, Caroline C G; Hedwig, Berthold; Conduit, Graham; Lawrence, Peter A; Goodwin, Stephen F; Casal, José

    2012-11-20

    Courtship in Drosophila melanogaster has become an iconic example of an innate and interactive series of behaviors. The female signals her acceptance of copulation by becoming immobile in response to a male's display of stereotyped actions. The male and female communicate via vision, air-borne sounds, and pheromones, but what triggers the female's immobility is undetermined. Here, we describe an overlooked and important component of Drosophila courtship. Video recordings and laser vibrometry show that the male abdomen shakes ("quivers"), generating substrate-borne vibrations at about six pulses per second. We present evidence that the female becomes receptive and stops walking because she senses these vibrations, rather than as a response to air-borne songs produced by the male fluttering the wings. We also present evidence that the neural circuits expressing the sex-determination genes fruitless and doublesex drive quivering behavior. These abdominal quivers and associated vibrations, as well as their effect on female receptivity, are conserved in other Drosophila species. Substrate-borne vibrations are an ancient form of communication that is widespread in animals. Our findings in Drosophila open a door to study the neuromuscular circuitry responsible for these signals and the sensory systems needed for their reception. PMID:23103187

  15. Unraveling Selection in the Mitochondrial Genome of Drosophila

    PubMed Central

    Ballard, JWO.; Kreitman, M.

    1994-01-01

    We examine mitochondrial DNA variation at the cytochrome b locus within and between three species of Drosophila to determine whether patterns of variation conform to the predictions of neutral molecular evolution. The entire 1137-bp cytochrome b locus was sequenced in 16 lines of Drosophila melanogaster, 18 lines of Drosophila simulans and 13 lines of Drosophila yakuba. Patterns of variation depart from neutrality by several test criteria. Analysis of the evolutionary clock hypothesis shows unequal rates of change along D. simulans lineages. A comparison within and between species of the ratio of amino acid replacement change to synonymous change reveals a relative excess of amino acid replacement polymorphism compared to the neutral prediction, suggestive of slightly deleterious or diversifying selection. There is evidence for excess homozygosity in our world wide sample of D. melanogaster and D. simulans alleles, as well as a reduction in the number of segregating sites in D. simulans, indicative of selective sweeps. Furthermore, a test of neutrality for codon usage shows the direction of mutations at third positions differs among different topological regions of the gene tree. The analyses indicate that molecular variation and evolution of mtDNA are governed by many of the same selective forces that have been shown to govern nuclear genome evolution and suggest caution be taken in the use of mtDNA as a ``neutral'' molecular marker. PMID:7851772

  16. Sipunculans and segmentation

    PubMed Central

    Kristof, Alen; Brinkmann, Nora

    2009-01-01

    Comparative molecular, developmental and morphogenetic analyses show that the three major segmented animal groups—Lophotrochozoa, Ecdysozoa and Vertebrata—use a wide range of ontogenetic pathways to establish metameric body organization. Even in the life history of a single specimen, different mechanisms may act on the level of gene expression, cell proliferation, tissue differentiation and organ system formation in individual segments. Accordingly, in some polychaete annelids the first three pairs of segmental peripheral neurons arise synchronously, while the metameric commissures of the ventral nervous system form in anterior-posterior progression. Contrary to traditional belief, loss of segmentation may have occurred more often than commonly assumed, as exemplified in the sipunculans, which show remnants of segmentation in larval stages but are unsegmented as adults. The developmental plasticity and potential evolutionary lability of segmentation nourishes the controversy of a segmented bilaterian ancestor versus multiple independent evolution of segmentation in respective metazoan lineages. PMID:19513266

  17. Segmented trapped vortex cavity

    NASA Technical Reports Server (NTRS)

    Grammel, Jr., Leonard Paul (Inventor); Pennekamp, David Lance (Inventor); Winslow, Jr., Ralph Henry (Inventor)

    2010-01-01

    An annular trapped vortex cavity assembly segment comprising includes a cavity forward wall, a cavity aft wall, and a cavity radially outer wall there between defining a cavity segment therein. A cavity opening extends between the forward and aft walls at a radially inner end of the assembly segment. Radially spaced apart pluralities of air injection first and second holes extend through the forward and aft walls respectively. The segment may include first and second expansion joint features at distal first and second ends respectively of the segment. The segment may include a forward subcomponent including the cavity forward wall attached to an aft subcomponent including the cavity aft wall. The forward and aft subcomponents include forward and aft portions of the cavity radially outer wall respectively. A ring of the segments may be circumferentially disposed about an axis to form an annular segmented vortex cavity assembly.

  18. [Bilateral segmental neurofibromatosis].

    PubMed

    Rose, I; Vakilzadeh, F

    1991-12-01

    Segmental neurofibromatosis is a rare type of neurofibromatosis. We report a case of bilateral manifestation, review the literature on this extremely uncommon variant, and discuss the possible causative mechanisms and the genetic risk of segmental neurofibromatosis. PMID:1765491

  19. Station Tour: Russian Segment

    NASA Video Gallery

    Expedition 33 Commander Suni Williams concludes her tour of the International Space Station with a visit to the Russian segment, which includes Zarya, the first segment of the station launched in 1...

  20. Ultrastructural correlates of left ventricular contraction abnormalities in patients with chronic ischemic heart disease: determinants of reversible segmental asynergy postrevascularization surgery.

    PubMed

    Flameng, W; Suy, R; Schwarz, F; Borgers, M; Piessens, J; Thone, F; Van Ermen, H; De Geest, H

    1981-11-01

    The relationships between structural alterations and left ventricular (LV) contraction abnormalities were studied in patients with coronary artery disease (CAD). Transmural biopsies of the LV anterior free wall were taken during aortocoronary bypass surgery (CABG) in 62 patients. When preoperative anterior wall motion (AWM) was reduced, significant myocardial cell degeneration was found in patients with as well as without previous anterior infarction (MI). The amount of myocardial fibrosis was increased only in patients with ECG evidence of previous anterior MI (p less than 0.001). In a second series of 139 CAD patients, cineventriculograms performed before and 8 months after CABG were examined. In patients with patent grafts to the LV anterior wall not previously infarcted, reduced AWM became normal. In patients with previous anterior MI the outcome of AWM was unpredictable (usually unimproved). Thus the histologic correlate of reduced AWM in segments not previously infarcted was progressive loss of contractile material in otherwise viable myocardial cells. Some reversibility was suggested by restoration of resting function after CABG. Unpredictable results in segments associated with pathologic Q waves appear related to the fibrous component of these previously infarcted areas. PMID:6975559

  1. Turing-Child field underlies spatial periodicity in Drosophila and planarians.

    PubMed

    Schiffmann, Yoram

    2011-05-01

    The regular spatial periodicity manifested in Drosophila gene expression has been considered as a test case for the involvement of a Turing system in biology. It was expected--if such involvement exists--to find a spatially periodic protein distribution where the proteins are Turing morphogens. The failure to find such a periodic distribution of Turing proteins, and the experimental findings of the involvement of different combinations of regulatory proteins and different binding sites for the different stripes of a periodic gene expression, has resulted in the dismissal of the involvement of a Turing system in Drosophila periodicity and segmentation. But if one is willing to allow a Turing system in the level of post-translational modification of proteins instead of in the protein level, one can explain the regular spatial periodicity of gene expression. The source of the spatial periodicity of gene expression does not lie in the regulatory proteins, but in the spatially periodic post-translational modification of these broadly distributed upstream regulatory proteins. The post-translational modification provides the missing spatial information for the regular pattern of 14 stripes. We report that such a field with segmental spatial periodicity that can affect downstream proteins and modify them post-translationally and periodically has been observed. This is the Turing-Child (TC) field. We explain the recent observation in Drosophila of phosphorylated transcription factor distributed with segmental periodicity, the disappearance of the spatially periodic gene expression when the regulatory protein loses its normal ability to be phosphorylated, and the spatially periodic segmental groove formation. Just as the reduction of Turing wavelength causes the appearance of 14 stripes in Drosophila so it causes the appearance of bipolar 2-headed Planaria. PMID:21187110

  2. Segmented Target Design

    NASA Astrophysics Data System (ADS)

    Merhi, Abdul Rahman; Frank, Nathan; Gueye, Paul; Thoennessen, Michael; MoNA Collaboration

    2013-10-01

    A proposed segmented target would improve decay energy measurements of neutron-unbound nuclei. Experiments like this have been performed at the National Superconducting Cyclotron Laboratory (NSCL) located at Michigan State University. Many different nuclei are produced in such experiments, some of which immediately decay into a charged particle and neutron. The charged particles are bent by a large magnet and measured by a suite of charged particle detectors. The neutrons are measured by the Modular Neutron Array (MoNA) and Large Multi-Institutional Scintillation Array (LISA). With the current target setup, a nucleus in a neutron-unbound state is produced with a radioactive beam impinged upon a beryllium target. The resolution of these measurements is very dependent on the target thickness since the nuclear interaction point is unknown. In a segmented target using alternating layers of silicon detectors and Be-targets, the Be-target in which the nuclear reaction takes place would be determined. Thus the experimental resolution would improve. This poster will describe the improvement over the current target along with the status of the design. Work supported by Augustana College and the National Science Foundation grant #0969173.

  3. Possible and Impossible Segments.

    ERIC Educational Resources Information Center

    Walker, Rachel; Pullum, Geoffrey K.

    1999-01-01

    Examines the relationship between phonetic possibility and phonological permissibility of segment types. Specific focus is on whether there are any phonetically impossible segments phonologically permissible, and whether there are any phonetically possible segments phonologically impermissable. Examines the case of nasality spreading in Sudanese…

  4. MUTAGENESIS SCREENING OF PESTICIDES 'DROSOPHILA'

    EPA Science Inventory

    Drosophila melanogaster males were exposed by feeding (plus contact and possibly inhalation). The genetic test found most sensitive and appropriate was the sex-linked recessive lethal test. For this, males of the Canton-S wild type stock were exposed. They were mated individually...

  5. A DNA Virus of Drosophila

    PubMed Central

    Unckless, Robert L.

    2011-01-01

    Little is known about the viruses infecting most species. Even in groups as well-studied as Drosophila, only a handful of viruses have been well-characterized. A viral metagenomic approach was used to explore viral diversity in 83 wild-caught Drosophila innubila, a mushroom feeding member of the quinaria group. A single fly that was injected with, and died from, Drosophila C Virus (DCV) was added to the sample as a control. Two-thirds of reads in the infected sample had DCV as the best BLAST hit, suggesting that the protocol developed is highly sensitive. In addition to the DCV hits, several sequences had Oryctes rhinoceros Nudivirus, a double-stranded DNA virus, as a best BLAST hit. The virus associated with these sequences was termed Drosophila innubila Nudivirus (DiNV). PCR screens of natural populations showed that DiNV was both common and widespread taxonomically and geographically. Electron microscopy confirms the presence of virions in fly fecal material similar in structure to other described Nudiviruses. In 2 species, D. innubila and D. falleni, the virus is associated with a severe (∼80–90%) loss of fecundity and significantly decreased lifespan. PMID:22053195

  6. Antibody Staining in Drosophila Germaria.

    PubMed

    Lie-Jensen, Anette; Haglund, Kaisa

    2016-01-01

    Drosophila oogenesis is a powerful model for studying a wide spectrum of cellular and developmental processes in vivo. Oogenesis starts in a specialized structure called the germarium, which harbors the stem cells for both germ and somatic cells. The germarium produces egg chambers, each of which will develop into an egg. Active areas of research in Drosophila germaria include stem cell self-renewal, division, and maintenance, cell cycle control and differentiation, oocyte specification, intercellular communication, and signaling, among others. The solid knowledge base, the genetic tractability of the Drosophila model, as well as the availability and fast development of tools and imaging techniques for oogenesis research ensure that studies in this model will keep being instrumental for novel discoveries within cell and developmental biology also in the future. This chapter focuses on antibody staining in Drosophila germaria and provides a protocol for immunostaining as well as an overview of commonly used antibodies for visualization of different cell types and cellular structures. The protocol is well-suited for subsequent confocal microscopy analyses, and in addition we present key adaptations of the protocol that are useful when performing structured illumination microscopy (SIM) super-resolution imaging. PMID:27557571

  7. Iron Absorption in Drosophila melanogaster

    PubMed Central

    Mandilaras, Konstantinos; Pathmanathan, Tharse; Missirlis, Fanis

    2013-01-01

    The way in which Drosophila melanogaster acquires iron from the diet remains poorly understood despite iron absorption being of vital significance for larval growth. To describe the process of organismal iron absorption, consideration needs to be given to cellular iron import, storage, export and how intestinal epithelial cells sense and respond to iron availability. Here we review studies on the Divalent Metal Transporter-1 homolog Malvolio (iron import), the recent discovery that Multicopper Oxidase-1 has ferroxidase activity (iron export) and the role of ferritin in the process of iron acquisition (iron storage). We also describe what is known about iron regulation in insect cells. We then draw upon knowledge from mammalian iron homeostasis to identify candidate genes in flies. Questions arise from the lack of conservation in Drosophila for key mammalian players, such as ferroportin, hepcidin and all the components of the hemochromatosis-related pathway. Drosophila and other insects also lack erythropoiesis. Thus, systemic iron regulation is likely to be conveyed by different signaling pathways and tissue requirements. The significance of regulating intestinal iron uptake is inferred from reports linking Drosophila developmental, immune, heat-shock and behavioral responses to iron sequestration. PMID:23686013

  8. Iron absorption in Drosophila melanogaster.

    PubMed

    Mandilaras, Konstantinos; Pathmanathan, Tharse; Missirlis, Fanis

    2013-05-01

    The way in which Drosophila melanogaster acquires iron from the diet remains poorly understood despite iron absorption being of vital significance for larval growth. To describe the process of organismal iron absorption, consideration needs to be given to cellular iron import, storage, export and how intestinal epithelial cells sense and respond to iron availability. Here we review studies on the Divalent Metal Transporter-1 homolog Malvolio (iron import), the recent discovery that Multicopper Oxidase-1 has ferroxidase activity (iron export) and the role of ferritin in the process of iron acquisition (iron storage). We also describe what is known about iron regulation in insect cells. We then draw upon knowledge from mammalian iron homeostasis to identify candidate genes in flies. Questions arise from the lack of conservation in Drosophila for key mammalian players, such as ferroportin, hepcidin and all the components of the hemochromatosis-related pathway. Drosophila and other insects also lack erythropoiesis. Thus, systemic iron regulation is likely to be conveyed by different signaling pathways and tissue requirements. The significance of regulating intestinal iron uptake is inferred from reports linking Drosophila developmental, immune, heat-shock and behavioral responses to iron sequestration. PMID:23686013

  9. Drosophila Photoreceptors and Signaling Mechanisms

    PubMed Central

    Katz, Ben; Minke, Baruch

    2009-01-01

    Fly eyes have been a useful biological system in which fundamental principles of sensory signaling have been elucidated. The physiological optics of the fly compound eye, which was discovered in the Musca, Calliphora and Drosophila flies, has been widely exploited in pioneering genetic and developmental studies. The detailed photochemical cycle of bistable photopigments has been elucidated in Drosophila using the genetic approach. Studies of Drosophila phototransduction using the genetic approach have led to the discovery of novel proteins crucial to many biological processes. A notable example is the discovery of the inactivation no afterpotential D scaffold protein, which binds the light-activated channel, its activator the phospholipase C and it regulator protein kinase C. An additional protein discovered in the Drosophila eye is the light-activated channel transient receptor potential (TRP), the founding member of the diverse and widely spread TRP channel superfamily. The fly eye has thus played a major role in the molecular identification of processes and proteins with prime importance. PMID:19623243

  10. Antioxidants, metabolic rate and aging in Drosophila

    NASA Technical Reports Server (NTRS)

    Miquel, J.; Fleming, J.; Economos, A. C.

    1982-01-01

    The metabolic rate-of-living theory of aging was investigated by determining the effect of several life-prolonging antioxidants on the metabolic rate and life span of Drosophila. The respiration rate of groups of continuously agitated flies was determined in a Gilson respirometer. Vitamin E, 2,4-dinitrophenol, nordihydroguaiaretic acid, and thiazolidine carboxylic acid were employed as antioxidants. Results show that all of these antioxidants reduced the oxygen consumption rate and increased the mean life span, and a significant negative linear correlation was found between the mean life span and the metabolic rate. It is concluded that these findings indicate that some antioxidants may inhibit respiration rate in addition to their protective effect against free radical-induced cellular damage.

  11. Mechanisms and constraints shaping the evolution of body plan segmentation.

    PubMed

    Ten Tusscher, K H W J

    2013-05-01

    Segmentation of the major body axis into repeating units is arguably one of the major inventions in the evolution of animal body plan pattering. It is found in current day vertebrates, annelids and arthropods. Most segmented animals seem to use a clock-and-wavefront type mechanism in which oscillations emanating from a posterior growth zone become transformed into an anterior posterior sequence of segments. In contrast, few animals such as Drosophila use a complex gene regulatory hierarchy to simultaneously subdivide their entire body axis into segments. Here I discuss how in silico models simulating the evolution of developmental patterning can be used to investigate the forces and constraints that helped shape these two developmental modes. I perform an analysis of a series of previous simulation studies, exploiting the similarities and differences in their outcomes in relation to model characteristics to elucidate the circumstances and constraints likely to have been important for the evolution of sequential and simultaneous segmentation modes. The analysis suggests that constraints arising from the involved growth process and spatial patterning signal--posterior elongation producing a propagating wavefront versus a tissue wide morphogen gradient--and the evolutionary history--ancestral versus derived segmentation mode--strongly shaped both segmentation mechanisms. Furthermore, this implies that these patterning types are to be expected rather than random evolutionary outcomes and supports the likelihood of multiple parallel evolutionary origins. PMID:23708840

  12. Formation of the insect head involves lateral contribution of the intercalary segment, which depends on Tc-labial function.

    PubMed

    Posnien, Nico; Bucher, Gregor

    2010-02-01

    The insect head is composed of several segments. During embryonic development, the segments fuse to form a rigid head capsule where obvious segmental boundaries are lacking. Hence, the assignment of regions of the insect head to specific segments is hampered, especially with respect to dorsal (vertex) and lateral (gena) parts. We show that upon Tribolium labial (Tc-lab) knock down, the intercalary segment is deleted but not transformed. Furthermore, we find that the intercalary segment contributes to lateral parts of the head cuticle in Tribolium. Based on several additional mutant and RNAi phenotypes that interfere with gnathal segment development, we show that these segments do not contribute to the dorsal head capsule apart from the dorsal ridge. Opposing the classical view but in line with findings in the vinegar fly Drosophila melanogaster and the milkweed bug Oncopeltus fasciatus, we propose a "bend and zipper" model for insect head capsule formation. PMID:19913530

  13. P element homing to the Drosophila bithorax complex.

    PubMed

    Bender, W; Hudson, A

    2000-09-01

    P elements containing a 7 kb DNA fragment from the middle of the Drosophila bithorax complex insert preferentially into the bithorax complex or into the adjacent chromosome regions. This 'homing' property is similar to that reported for the engrailed promoter (Hama, C., Ali, Z. and Kornberg, T. B. (1990) Genes Dev. 4, 1079-1093). The 7 kb fragment does not contain any known promoter, but it acts as a boundary element separating adjacent segmental domains. An enhancer-trap P element was constructed with the homing fragment and the selectable marker flanked by FRT sites. P insertions can be trimmed down by Flp-mediated recombination to just the lacZ reporter, so that the (beta)-galactosidase pattern is not influenced by sequences inside the P element. Twenty insertions into the bithorax complex express (beta)-galactosidase in segmentally limited patterns, reflecting the segmental domains of the bithorax complex where the elements reside. The mapping of segmental domains has now been revised, with enlargement of the abx/bx, bxd/pbx, and the iab-3 domains. The FRT sites in the P elements permit recombination between pairs of elements on opposite chromosomes, to generate duplications or deletions of the DNA between the two insertion sites. Using this technique, the length of the Ultrabithorax transcription unit was varied from 37 to 138 kb, but there was surprisingly little effect on Ultrabithorax function. PMID:10952896

  14. Labor Market Segmentation and Librarian Salaries.

    ERIC Educational Resources Information Center

    Van House, Nancy A.

    1987-01-01

    Segmented labor market theory is used to explain how the structure of the library labor market may determine salary differences by type of library. Evidence that segmentation exists at intraoccupational levels and the possibility that comparing entire occupations may obscure results are also reported. (Author/CLB)

  15. Probes of chromatin accessibility in the Drosophila bithorax complex respond differently to Polycomb-mediated repression.

    PubMed Central

    McCall, K; Bender, W

    1996-01-01

    The Polycomb group (PcG) of genes are required for maintenance of the repressed state of the homeotic genes in Drosophila. There are similarities between the PcG repression and mating-type silencing in yeast or heterochromatic position effect in Drosophila, which suggest that PcG repression may involve a highly compacted chromatin structure. To test for such a structure, heterologous DNA- binding proteins were used as probes for DNA accessibility in Drosophila embryos. Binding sites for the yeast transcriptional activator GAL4 and for bacteriophage T7 RNA polymerase were inserted into the bithorax (bx) regulatory region of the endogenous Ultrabithorax (Ubx) gene, which is regulated by the PcG. Ubiquitously expressed GAL4 protein directs transcription through its binding sites only in the posterior segments where the bx region is active. The block to GAL4 activation in the more anterior segments is dependent on Polycomb (Pc) function. In contrast, T7 RNA polymerase can transcribe from its target promoter in all segments of the embryo. Thus, Pc-mediated repression blocks activated polymerase II transcription, but does not simply exclude all proteins. Images PMID:8599940

  16. Distribution Metrics and Image Segmentation

    PubMed Central

    Georgiou, Tryphon; Michailovich, Oleg; Rathi, Yogesh; Malcolm, James; Tannenbaum, Allen

    2007-01-01

    The purpose of this paper is to describe certain alternative metrics for quantifying distances between distributions, and to explain their use and relevance in visual tracking. Besides the theoretical interest, such metrics may be used to design filters for image segmentation, that is for solving the key visual task of separating an object from the background in an image. The segmenting curve is represented as the zero level set of a signed distance function. Most existing methods in the geometric active contour framework perform segmentation by maximizing the separation of intensity moments between the interior and the exterior of an evolving contour. Here one can use the given distributional metric to determine a flow which minimizes changes in the distribution inside and outside the curve. PMID:18769529

  17. Evolutionary Origin of Body Axis Segmentation in Annelids and Arthropods

    NASA Technical Reports Server (NTRS)

    Shankland, S. Martin

    2003-01-01

    During the period of this report, we have made a number of important discoveries. To date this work has led to 4 peer-reviewed publications in primary research journals plus 1 minireview and 1 chapter in the proceedings of a meeting. Publications resulting from this grant support are enumerated at the end of the report. Two additional, on-going studies also described. 1. Using laser cell ablation, we have obtained evidence that an annelid - the leech Helobdella robusta - patterns the anteroposterior (AP) polarity of its nascent segment primordia independent of cell interactions oriented along the AP axis. 2. We cloned a Helobdella homologue (hro-hh) of the Drosophila segment polarity gene hedgehog, and used in situ hybridization and northern blots to characterize its expression in the embryo. 3. We have used laser cell ablations to examine the possible role of cell interactions during the developmental patterning of the 4 rostralmost "head" segments of the leech Helobdella robusta.

  18. Multi-segment detector

    NASA Technical Reports Server (NTRS)

    George, Peter K. (Inventor)

    1978-01-01

    A plurality of stretcher detector segments are connected in series whereby detector signals generated when a bubble passes thereby are added together. Each of the stretcher detector segments is disposed an identical propagation distance away from passive replicators wherein bubbles are replicated from a propagation path and applied, simultaneously, to the stretcher detector segments. The stretcher detector segments are arranged to include both dummy and active portions thereof which are arranged to permit the geometry of both the dummy and active portions of the segment to be substantially matched.

  19. A Systematic Cell-Based Analysis of Localization of Predicted Drosophila Peroxisomal Proteins.

    PubMed

    Baron, Matthew N; Klinger, Christen M; Rachubinski, Richard A; Simmonds, Andrew J

    2016-05-01

    Peroxisomes are membrane-bound organelles found in almost all eukaryotic cells. They perform specialized biochemical functions that vary with organism, tissue or cell type. Mutations in human genes required for the assembly of peroxisomes result in a spectrum of diseases called the peroxisome biogenesis disorders. A previous sequence-based comparison of the predicted proteome of Drosophila melanogaster (the fruit fly) to human proteins identified 82 potential homologues of proteins involved in peroxisomal biogenesis, homeostasis or metabolism. However, the subcellular localization of these proteins relative to the peroxisome was not determined. Accordingly, we tested systematically the localization and selected functions of epitope-tagged proteins in Drosophila Schneider 2 cells to determine the subcellular localization of 82 potential Drosophila peroxisomal protein homologues. Excluding the Pex proteins, 34 proteins localized primarily to the peroxisome, 8 showed dual localization to the peroxisome and other structures, and 26 localized exclusively to organelles other than the peroxisome. Drosophila is a well-developed laboratory animal often used for discovery of gene pathways, including those linked to human disease. Our work establishes a basic understanding of peroxisome protein localization in Drosophila. This will facilitate use of Drosophila as a genetically tractable, multicellular model system for studying key aspects of human peroxisome disease. PMID:26865094

  20. Heuristic segmentation of a nonstationary time series

    NASA Astrophysics Data System (ADS)

    Fukuda, Kensuke; Eugene Stanley, H.; Nunes Amaral, Luís A.

    2004-02-01

    Many phenomena, both natural and human influenced, give rise to signals whose statistical properties change under time translation, i.e., are nonstationary. For some practical purposes, a nonstationary time series can be seen as a concatenation of stationary segments. However, the exact segmentation of a nonstationary time series is a hard computational problem which cannot be solved exactly by existing methods. For this reason, heuristic methods have been proposed. Using one such method, it has been reported that for several cases of interest—e.g., heart beat data and Internet traffic fluctuations—the distribution of durations of these stationary segments decays with a power-law tail. A potential technical difficulty that has not been thoroughly investigated is that a nonstationary time series with a (scalefree) power-law distribution of stationary segments is harder to segment than other nonstationary time series because of the wider range of possible segment lengths. Here, we investigate the validity of a heuristic segmentation algorithm recently proposed by Bernaola-Galván et al. [Phys. Rev. Lett. 87, 168105 (2001)] by systematically analyzing surrogate time series with different statistical properties. We find that if a given nonstationary time series has stationary periods whose length is distributed as a power law, the algorithm can split the time series into a set of stationary segments with the correct statistical properties. We also find that the estimated power-law exponent of the distribution of stationary-segment lengths is affected by (i) the minimum segment length and (ii) the ratio R≡σɛ/σx¯, where σx¯ is the standard deviation of the mean values of the segments and σɛ is the standard deviation of the fluctuations within a segment. Furthermore, we determine that the performance of the algorithm is generally not affected by uncorrelated noise spikes or by weak long-range temporal correlations of the fluctuations within segments.

  1. Color image segmentation

    NASA Astrophysics Data System (ADS)

    McCrae, Kimberley A.; Ruck, Dennis W.; Rogers, Steven K.; Oxley, Mark E.

    1994-03-01

    The most difficult stage of automated target recognition is segmentation. Current segmentation problems include faces and tactical targets; previous efforts to segment these objects have used intensity and motion cues. This paper develops a color preprocessing scheme to be used with the other segmentation techniques. A neural network is trained to identify the color of a desired object, eliminating all but that color from the scene. Gabor correlations and 2D wavelet transformations will be performed on stationary images; and 3D wavelet transforms on multispectral data will incorporate color and motion detection into the machine visual system. The paper will demonstrate that color and motion cues can enhance a computer segmentation system. Results from segmenting faces both from the AFIT data base and from video taped television are presented; results from tactical targets such as tanks and airplanes are also given. Color preprocessing is shown to greatly improve the segmentation in most cases.

  2. Method 353.4 Determination of Nitrate and Nitrite in Estuarine and Coastal Waters by Gas Segmented Continuous Flow Colorimetric Analysis

    EPA Science Inventory

    This method provides a procedure for determining nitrate and nitrite concentrations in estuarine and coastal waters. Nitrate is reduced to nitrite by cadmium,1-3 and the resulting nitrite determined by formation of an azo dye.4-6

  3. How Drosophila melanogaster Forms its Mechanoreceptors

    PubMed Central

    Furman, D.P; Bukharina, T.A

    2008-01-01

    A strictly determined number of external sensory organs, macrochaetes, acting as mechanoreceptors, are orderly located on drosophila head and body. Totally, they form the bristle pattern, which is a species-specific characteristic of drosophila. Each mechanoreceptor comprises four specialized cells derived from the single sensory organ precursor (SOP) cell. The conserved bristle pattern combined with a comparatively simple structure of each mechanosensory organ makes macrochaetes a convenient model for studying the formation spatial structures with a fixed number of elements at certain positions and the mechanism underlying cell differentiation. The macrochaete morphogenesis consists of three stages. At the first stage, the proneural clusters segregate from the massive of ectodermal cells of the wing imaginal disc. At the second stage, the SOP cell is determined and its position in the cluster is specified. At the third stage, the SOP cell undergoes two asymmetric divisions, and the daughter cells differentiate into the components of mechanoreceptor: shaft, socket, bipolar neuron, and sheath. The critical factor determining the neural pathway of cell development is the content of proneural proteins, products of the achaete-scute (AS-C) gene complex, reaching its maximum in the SOP cell. The experimental data on the main genes and their products involved in the control of bristle pattern formation are systematized. The roles of achaete-scute complex, EGFR and Notch signaling pathways, and selector genes in these processes are considered. An integral scheme describing the functioning of the system controlling macrochaete development in D. melanogaster is proposed based on analysis of literature data. PMID:19471605

  4. Development and organization of glial cells in Drosophila melanogaster.

    PubMed

    Giangrande, A

    1996-10-01

    Glial cells constitute a crucial component of the nervous system. They wrap the neuronal somata and axons and play a number of roles during normal neuronal development and activity as well as during axonal regeneration after wounding. The availability of cellular markers and genetic tools have made it possible in Drosophila to start identifying the genes and the cell-cell interactions leading to glial cell differentiation. The existence of multipotent precursor cells in the nervous system, the requirement for master genes determining the glial cell fate, the migratory abilities of fly glial cells and the existence of neuron-glial cell interactions during development are some of the features revealed by these approaches. These findings also indicate an evolutionary conservation in the developmental mechanisms between invertebrates and vertebrates. Finally, Drosophila is an ideal model system to determine in vivo the precise roles of glial cells and to study the etiology of pathologies associated with abnormal glial differentiation. PMID:8946240

  5. Non-destructive species identification of Drosophila obscura and D. subobscura (Diptera) using near-infrared spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The vinegar flies Drosophila subobscura and D. obscura frequently serve as study organisms for evolutionary biology. Their high morphological similarity renders traditional species determination difficult, especially when living specimens for setting up laboratory populations need to be identified. ...

  6. Functional Evolution of cis-Regulatory Modules at a Homeotic Gene in Drosophila

    PubMed Central

    Schiller, Benjamin J.; Bae, Esther; Tran, Diana A.; Shur, Andrey S.; Allen, John M.; Rau, Christoph; Bender, Welcome; Fisher, William W.; Celniker, Susan E.; Drewell, Robert A.

    2009-01-01

    It is a long-held belief in evolutionary biology that the rate of molecular evolution for a given DNA sequence is inversely related to the level of functional constraint. This belief holds true for the protein-coding homeotic (Hox) genes originally discovered in Drosophila melanogaster. Expression of the Hox genes in Drosophila embryos is essential for body patterning and is controlled by an extensive array of cis-regulatory modules (CRMs). How the regulatory modules functionally evolve in different species is not clear. A comparison of the CRMs for the Abdominal-B gene from different Drosophila species reveals relatively low levels of overall sequence conservation. However, embryonic enhancer CRMs from other Drosophila species direct transgenic reporter gene expression in the same spatial and temporal patterns during development as their D. melanogaster orthologs. Bioinformatic analysis reveals the presence of short conserved sequences within defined CRMs, representing gap and pair-rule transcription factor binding sites. One predicted binding site for the gap transcription factor KRUPPEL in the IAB5 CRM was found to be altered in Superabdominal (Sab) mutations. In Sab mutant flies, the third abdominal segment is transformed into a copy of the fifth abdominal segment. A model for KRUPPEL-mediated repression at this binding site is presented. These findings challenge our current understanding of the relationship between sequence evolution at the molecular level and functional activity of a CRM. While the overall sequence conservation at Drosophila CRMs is not distinctive from neighboring genomic regions, functionally critical transcription factor binding sites within embryonic enhancer CRMs are highly conserved. These results have implications for understanding mechanisms of gene expression during embryonic development, enhancer function, and the molecular evolution of eukaryotic regulatory modules. PMID:19893611

  7. kuzbanian-mediated cleavage of Drosophila Notch

    PubMed Central

    Lieber, Toby; Kidd, Simon; Young, Michael W.

    2002-01-01

    Loss of Kuzbanian, a member of the ADAM family of metalloproteases, produces neurogenic phenotypes in Drosophila. It has been suggested that this results from a requirement for kuzbanian-mediated cleavage of the Notch ligand Delta. Using transgenic Drosophila expressing transmembrane Notch proteins, we show that kuzbanian, independent of any role in Delta processing, is required for the cleavage of Notch. We show that Kuzbanian can physically associate with Notch and that removal of kuzbanian activity by RNA-mediated interference in Drosophila tissue culture cells eliminates processing of ligand-independent transmembrane Notch molecules. Our data suggest that in Drosophila, kuzbanian can mediate S2 cleavage of Notch. PMID:11799064

  8. A segmentation clock operating in blastoderm and germband stages of Tribolium development

    PubMed Central

    El-Sherif, Ezzat; Averof, Michalis; Brown, Susan J.

    2012-01-01

    In Drosophila, all segments form in the blastoderm where morphogen gradients spanning the entire anterior-posterior axis of the embryo provide positional information. However, in the beetle Tribolium castaneum and most other arthropods, a number of anterior segments form in the blastoderm, and the remaining segments form sequentially from a posterior growth zone during germband elongation. Recently, the cyclic nature of the pair-rule gene Tc-odd-skipped was demonstrated in the growth zone of Tribolium, indicating that a vertebrate-like segmentation clock is employed in the germband stage of its development. This suggests that two mechanisms might function in the same organism: a Drosophila-like mechanism in the blastoderm, and a vertebrate-like mechanism in the germband. Here, we show that segmentation at both blastoderm and germband stages of Tribolium is based on a segmentation clock. Specifically, we show that the Tribolium primary pair-rule gene, Tc-even-skipped (Tc-eve), is expressed in waves propagating from the posterior pole and progressively slowing until they freeze into stripes; such dynamics are a hallmark of clock-based segmentation. Phase shifts between Tc-eve transcripts and protein confirm that these waves are due to expression dynamics. Moreover, by tracking cells in live embryos and by analyzing mitotic profiles, we found that neither cell movement nor oriented cell division could explain the observed wave dynamics of Tc-eve. These results pose intriguing evolutionary questions, as Drosophila and Tribolium segment their blastoderms using the same genes but different mechanisms. PMID:23095886

  9. Structural determinants of alpha-bungarotoxin binding to the sequence segment 181-200 of the muscle nicotinic acetylcholine receptor. alpha. subunit: Effects of cysteine/cystine modification and species-specific amino acid substitution

    SciTech Connect

    McLane, K.E.; Wu, Xiadong; Diethelm, B.; Conti-Tronconi, B.M. )

    1991-05-21

    The sequence segment 181-200 of the Torpedo nicotinic acetylcholine receptor (nAChR) {alpha}subunit forms a binding site for {alpha}-bungarotoxin ({alpha}-BTX). Synthetic peptides corresponding to the homologous sequences of human, calf, mouse, chicken, frog, and cobra muscle nAChR {alpha}1 subunits were tested for their ability to bind {sup 125}I-{alpha}-BTX, and differences in {alpha}-BTX affinity were determined by using solution (IC{sub 50}s) and solid-phase (K{sub d}s) assays. Panels of overlapping peptides corresponding to the complete {alpha}1 subunit of mouse and human were also tested for {alpha}-BTX binding, but other sequence segments forming the {alpha}-BTX site were not consistently detectable. The role of a putative vicinal disulfide bound between Cys-192 and -193, relative to the Torpedo sequence, was determined by modifying the peptides with sulfhydryl reagents. Reduction and alkylation of the peptides decreased {alpha}-BTX binding, whereas oxidation of the peptides had little effect. These results indicate that while the adjacent cysteines are likely to be involved in forming the toxin/{alpha}1-subunit interface a vicinal disulfide bound was not required for {alpha}-BTX binding.

  10. Sensorimotor structure of Drosophila larva phototaxis

    PubMed Central

    Kane, Elizabeth A.; Gershow, Marc; Afonso, Bruno; Larderet, Ivan; Klein, Mason; Carter, Ashley R.; de Bivort, Benjamin L.; Sprecher, Simon G.; Samuel, Aravinthan D. T.

    2013-01-01

    The avoidance of light by fly larvae is a classic paradigm for sensorimotor behavior. Here, we use behavioral assays and video microscopy to quantify the sensorimotor structure of phototaxis using the Drosophila larva. Larval locomotion is composed of sequences of runs (periods of forward movement) that are interrupted by abrupt turns, during which the larva pauses and sweeps its head back and forth, probing local light information to determine the direction of the successive run. All phototactic responses are mediated by the same set of sensorimotor transformations that require temporal processing of sensory inputs. Through functional imaging and genetic inactivation of specific neurons downstream of the sensory periphery, we have begun to map these sensorimotor circuits into the larval central brain. We find that specific sensorimotor pathways that govern distinct light-evoked responses begin to segregate at the first relay after the photosensory neurons. PMID:24043822

  11. A Cluster of Cuticle Protein Genes of Drosophila Melanogaster at 65a: Sequence, Structure and Evolution

    PubMed Central

    Charles, J. P.; Chihara, C.; Nejad, S.; Riddiford, L. M.

    1997-01-01

    A 36-kb genomic DNA segment of the Drosophila melanogaster genome containing 12 clustered cuticle genes has been mapped and partially sequenced. The cluster maps at 65A 5-6 on the left arm of the third chromosome, in agreement with the previously determined location of a putative cluster encompassing the genes for the third instar larval cuticle proteins LCP5, LCP6 and LCP8. This cluster is the largest cuticle gene cluster discovered to date and shows a number of surprising features that explain in part the genetic complexity of the LCP5, LCP6 and LCP8 loci. The genes encoding LCP5 and LCP8 are multiple copy genes and the presence of extensive similarity in their coding regions gives the first evidence for gene conversion in cuticle genes. In addition, five genes in the cluster are intronless. Four of these five have arisen by retroposition. The other genes in the cluster have a single intron located at an unusual location for insect cuticle genes. PMID:9383064

  12. The Drosophila STUbL protein Degringolade limits HES functions during embryogenesis.

    PubMed

    Barry, Kevin C; Abed, Mona; Kenyagin, Dorit; Werwie, Timothy R; Boico, Olga; Orian, Amir; Parkhurst, Susan M

    2011-05-01

    Degringolade (Dgrn) encodes a Drosophila SUMO-targeted ubiquitin ligase (STUbL) protein similar to that of mammalian RNF4. Dgrn facilitates the ubiquitylation of the HES protein Hairy, which disrupts the repressive activity of Hairy by inhibiting the recruitment of its cofactor Groucho. We show that Hey and all HES family members, except Her, interact with Dgrn and are substrates for its E3 ubiquitin ligase activity. Dgrn displays dynamic subcellular localization, accumulates in the nucleus at times when HES family members are active and limits Hey and HES family activity during sex determination, segmentation and neurogenesis. We show that Dgrn interacts with the Notch signaling pathway by it antagonizing the activity of E(spl)-C proteins. dgrn null mutants are female sterile, producing embryos that arrest development after two or three nuclear divisions. These mutant embryos exhibit fragmented or decondensed nuclei and accumulate higher levels of SUMO-conjugated proteins, suggesting a role for Dgrn in genome stability. PMID:21486924

  13. Interspecific transplantation of polar plasm between Drosophila embryos

    PubMed Central

    1976-01-01

    Posterior polar plasm of the Drosophila egg has been shown to function autonomously in germ cell determination after transplantation to either the anterior or mid-ventral region of the early embryo. By means of similar transplantations, we have tested the ability of polar plasm of Drosophila immigrans to induce the formation of pole cells in a Drosophila melanogaster embryo. After the transplantation of polar plasm, "hybrid" pole cells were found in which both pole cell-specific organelles, the polar granules and nuclear body, were structurally similar to those characteristic of the transplanted cytoplasm. In order to determine whether these hybrid cells can function as germ cell precursors, these cells were transplanted to the posterior tip of genetically marked embryos. Approximately 5% of the flies obtained from embryos receiving potential pole cells produce offspring derived from the induced pole cells. This result demonstrates that polar plasm can function in interspecific species combinations and indicates that the molecular mechanisms of germ cell determination are conservative in evolution. Finally, in order to test whether there is any evidence for cytoplasmic inheritance of polar granules, embryos derived from hybrid pole cells were examined for their polar granule morphology. The fine structure of the granules conformed to that of the nucleus. Thus, no evidence was found for the cytoplasmic inheritance of these particular organelles. PMID:820700

  14. A Drosophila complementary DNA resource

    SciTech Connect

    Rubin, Gerald M.; Hong, Ling; Brokstein, Peter; Evans-Holm, Martha; Frise, Erwin; Stapleton, Mark; Harvey, Damon A.

    2000-03-24

    Collections of nonredundant, full-length complementary DNA (cDNA) clones for each of the model organisms and humans will be important resources for studies of gene structure and function. We describe a general strategy for producing such collections and its implementation, which so far has generated a set of cDNAs corresponding to over 40% of the genes in the fruit fly Drosophila melanogaster.

  15. Impact assisted segmented cutterhead

    DOEpatents

    Morrell, Roger J.; Larson, David A.; Ruzzi, Peter L.

    1992-01-01

    An impact assisted segmented cutterhead device is provided for cutting various surfaces from coal to granite. The device comprises a plurality of cutting bit segments deployed in side by side relationship to form a continuous cutting face and a plurality of impactors individually associated with respective cutting bit segments. An impactor rod of each impactor connects that impactor to the corresponding cutting bit segment. A plurality of shock mounts dampening the vibration from the associated impactor. Mounting brackets are used in mounting the cutterhead to a base machine.

  16. Optogenetic pacing in Drosophila melanogaster

    PubMed Central

    Alex, Aneesh; Li, Airong; Tanzi, Rudolph E.; Zhou, Chao

    2015-01-01

    Electrical stimulation is currently the gold standard for cardiac pacing. However, it is invasive and nonspecific for cardiac tissues. We recently developed a noninvasive cardiac pacing technique using optogenetic tools, which are widely used in neuroscience. Optogenetic pacing of the heart provides high spatial and temporal precisions, is specific for cardiac tissues, avoids artifacts associated with electrical stimulation, and therefore promises to be a powerful tool in basic cardiac research. We demonstrated optogenetic control of heart rhythm in a well-established model organism, Drosophila melanogaster. We developed transgenic flies expressing a light-gated cation channel, channelrhodopsin-2 (ChR2), specifically in their hearts and demonstrated successful optogenetic pacing of ChR2-expressing Drosophila at different developmental stages, including the larva, pupa, and adult stages. A high-speed and ultrahigh-resolution optical coherence microscopy imaging system that is capable of providing images at a rate of 130 frames/s with axial and transverse resolutions of 1.5 and 3.9 μm, respectively, was used to noninvasively monitor Drosophila cardiac function and its response to pacing stimulation. The development of a noninvasive integrated optical pacing and imaging system provides a novel platform for performing research studies in developmental cardiology. PMID:26601299

  17. Leigh Syndrome in Drosophila melanogaster

    PubMed Central

    Da-Rè, Caterina; von Stockum, Sophia; Biscontin, Alberto; Millino, Caterina; Cisotto, Paola; Zordan, Mauro A.; Zeviani, Massimo; Bernardi, Paolo; De Pittà, Cristiano; Costa, Rodolfo

    2014-01-01

    Leigh Syndrome (LS) is the most common early-onset, progressive mitochondrial encephalopathy usually leading to early death. The single most prevalent cause of LS is occurrence of mutations in the SURF1 gene, and LSSurf1 patients show a ubiquitous and specific decrease in the activity of mitochondrial respiratory chain complex IV (cytochrome c oxidase, COX). SURF1 encodes an inner membrane mitochondrial protein involved in COX assembly. We established a Drosophila melanogaster model of LS based on the post-transcriptional silencing of CG9943, the Drosophila homolog of SURF1. Knockdown of Surf1 was induced ubiquitously in larvae and adults, which led to lethality; in the mesodermal derivatives, which led to pupal lethality; or in the central nervous system, which allowed survival. A biochemical characterization was carried out in knockdown individuals, which revealed that larvae unexpectedly displayed defects in all complexes of the mitochondrial respiratory chain and in the F-ATP synthase, while adults had a COX-selective impairment. Silencing of Surf1 expression in Drosophila S2R+ cells led to selective loss of COX activity associated with decreased oxygen consumption and respiratory reserve. We conclude that Surf1 is essential for COX activity and mitochondrial function in D. melanogaster, thus providing a new tool that may help clarify the pathogenic mechanisms of LS. PMID:25164807

  18. Insulin receptor in Drosophila melanogaster

    SciTech Connect

    Petruzzelli, L.; Herrera, R.; Rosen, O.

    1986-05-01

    A specific, high affinity insulin receptor is present in both adult Drosophila and in Drosophila embryos. Wheat germ lectin-enriched extracts of detergent-solubilized membranes from embryos and adults bind insulin with a K/sub d/ of 15 nM. Binding is specific for insulin; micromolar concentrations of proinsulin, IGFI, and IGFII are required to displace bound /sup 125/I-insulin. Insulin-dependent protein tyrosine kinase activity appears during embryogenesis. It is evident between 6 and 12 hours of development, peaks between 12 and 18 hours and falls in the adult. During 0-6 hours of embryogenesis, and in the adult, a specific protein band (Mr = 135,000) is crosslinked to /sup 125/I-insulin. During 6-12 and 12-18 hours of embryogenesis stages in which insulin-dependent protein tyrosine kinase is high, an additional band (Mr = 100,000) becomes crosslinked to /sup 125/I-insulin. Isolation and DNA sequence analysis of genomic clones encoding the Drosophila insulin receptor will be presented as will the characterization of insulin receptor mRNA's during development.

  19. 'Peer pressure' in larval Drosophila?

    PubMed

    Niewalda, Thomas; Jeske, Ines; Michels, Birgit; Gerber, Bertram

    2014-01-01

    Understanding social behaviour requires a study case that is simple enough to be tractable, yet complex enough to remain interesting. Do larval Drosophila meet these requirements? In a broad sense, this question can refer to effects of the mere presence of other larvae on the behaviour of a target individual. Here we focused in a more strict sense on 'peer pressure', that is on the question of whether the behaviour of a target individual larva is affected by what a surrounding group of larvae is doing. We found that innate olfactory preference of a target individual was neither affected (i) by the level of innate olfactory preference in the surrounding group nor (ii) by the expression of learned olfactory preference in the group. Likewise, learned olfactory preference of a target individual was neither affected (iii) by the level of innate olfactory preference of the surrounding group nor (iv) by the learned olfactory preference the group was expressing. We conclude that larval Drosophila thus do not take note of specifically what surrounding larvae are doing. This implies that in a strict sense, and to the extent tested, there is no social interaction between larvae. These results validate widely used en mass approaches to the behaviour of larval Drosophila. PMID:24907371

  20. The involvement of engrailed and wingless during segmentation in the onychophoran Euperipatoides kanangrensis (Peripatopsidae: Onychophora) (Reid 1996).

    PubMed

    Eriksson, Bo Joakim; Tait, Noel N; Budd, Graham E; Akam, Michael

    2009-05-01

    As the putative sister group to the arthropods, onychophorans can provide insight into ancestral developmental mechanisms in the panarthropod clade. Here, we examine the expression during segmentation of orthologues of wingless (Wnt1) and engrailed, two genes that play a key role in defining segment boundaries in Drosophila and that appear to play a role in segmentation in many other arthropods. Both are expressed in segmentally reiterated stripes in all forming segments except the first (brain) segment, which only shows an engrailed stripe. Engrailed is expressed before segments are morphologically visible and is expressed in both mesoderm and ectoderm. Segmental wingless expression is not detectable until after mesodermal somites are clearly distinct. Early engrailed expression lies in and extends to both sides of the furrow that first demarcates segments in the ectoderm, but is largely restricted to the posterior part of somites. Wingless expression lies immediately anterior to engrailed expression, as it does in many arthropods, but there is no precise cellular boundary between the two expression domains analogous to the overt parasegment boundary seen in Drosophila. Engrailed stripes extend along the posterior part of each limb bud, including the antenna, while wingless is restricted to the distal tip of the limbs and the neurectoderm basal to the limbs. PMID:19434423

  1. Fluorescent visualization of macromolecules in Drosophila whole mounts.

    PubMed

    Ramos, Ricardo Guelerman Pinheiro; Machado, Luciana Claudia Herculano; Moda, Livia Maria Rosatto

    2010-01-01

    The ability to determine the expression dynamics of individual genes "in situ" by visualizing the precise spatial and temporal distribution of their products in whole mounts by histochemical and immunocytochemical reactions has revolutionized our understanding of cellular processes. Drosophila developmental genetics was one of the fields that benefited most from these technologies, and a variety of fluorescent methods were specifically designed for investigating the localization of developmentally important proteins and cell markers during embryonic and post embryonic stages of this model organism. In this chapter we present detailed protocols for fluorescence immunocytochemistry of whole mount embryos, imaginal discs, pupal retinas, and salivary glands of Drosophila melanogaster, as well as methods for fluorescent visualization of specific subcellular structures in these tissues. PMID:20012830

  2. Clonal development and organization of the adult Drosophila central brain

    PubMed Central

    Yu, Hung-Hsiang; Awasaki, Takeshi; Schroeder, Mark David; Long, Fuhui; Yang, Jacob S.; He, Yisheng; Ding, Peng; Kao, Jui-Chun; Wu, Gloria Yueh-Yi; Peng, Hanchuan; Myers, Gene; Lee, Tzumin

    2013-01-01

    Summary Background The insect brain can be divided into neuropils that are formed by neurites of both local and remote origin. The complexity of the interconnections obscures how these neuropils are established and interconnected through development. The Drosophila central brain develops from a fixed number of neuroblasts (NBs) that deposit neurons in regional clusters. Results By determining individual NB clones and pursuing their projections into specific neuropils we unravel the regional development of the brain neural network. Exhaustive clonal analysis revealed 95 stereotyped neuronal lineages with characteristic cell body locations and neurite trajectories. Most clones show complex projection patterns, but despite the complexity, neighboring clones often co-innervate the same local neuropil(s) and further target a restricted set of distant neuropils. Conclusions These observations argue for regional clonal development of both neuropils and neuropil connectivity throughout the Drosophila central brain. PMID:23541733

  3. Structures of the human and Drosophila 80S ribosome.

    PubMed

    Anger, Andreas M; Armache, Jean-Paul; Berninghausen, Otto; Habeck, Michael; Subklewe, Marion; Wilson, Daniel N; Beckmann, Roland

    2013-05-01

    Protein synthesis in all cells is carried out by macromolecular machines called ribosomes. Although the structures of prokaryotic, yeast and protist ribosomes have been determined, the more complex molecular architecture of metazoan 80S ribosomes has so far remained elusive. Here we present structures of Drosophila melanogaster and Homo sapiens 80S ribosomes in complex with the translation factor eEF2, E-site transfer RNA and Stm1-like proteins, based on high-resolution cryo-electron-microscopy density maps. These structures not only illustrate the co-evolution of metazoan-specific ribosomal RNA with ribosomal proteins but also reveal the presence of two additional structural layers in metazoan ribosomes, a well-ordered inner layer covered by a flexible RNA outer layer. The human and Drosophila ribosome structures will provide the basis for more detailed structural, biochemical and genetic experiments. PMID:23636399

  4. Biology and physiology of Drosophila suzukii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spotted wing drosophila, Drosophila suzukii, quickly emerged as a devastating invasive pest of small and stone fruits in the Americas and Europe. To better understand the population dynamics of D. suzukii, we reviewed recent work on juvenile development, adult reproduction, and seasonal variation in...

  5. Drosophila and Beer: An Experimental Laboratory Exercise

    ERIC Educational Resources Information Center

    Kurvink, Karen

    2004-01-01

    Drosophila melanogaster is a popular organism for studying genetics and development. Maintaining Drosophila on medium prepared with varying concentrations of beer and evaluating the effects on reproduction, life cycle stages and other factors is one of the exercises that is versatile and applicable to many student levels.

  6. Antennal mechanosensory neurons mediate wing motor reflexes in flying Drosophila.

    PubMed

    Mamiya, Akira; Dickinson, Michael H

    2015-05-20

    Although many behavioral studies have shown the importance of antennal mechanosensation in various aspects of insect flight control, the identities of the mechanosensory neurons responsible for these functions are still unknown. One candidate is the Johnston's organ (JO) neurons that are located in the second antennal segment and detect phasic and tonic rotations of the third antennal segment relative to the second segment. To investigate how different classes of JO neurons respond to different types of antennal movement during flight, we combined 2-photon calcium imaging with a machine vision system to simultaneously record JO neuron activity and the antennal movement from tethered flying fruit flies (Drosophila melanogaster). We found that most classes of JO neurons respond strongly to antennal oscillation at the wing beat frequency, but not to the tonic deflections of the antennae. To study how flies use input from the JO neurons during flight, we genetically ablated specific classes of JO neurons and examined their effect on the wing motion. Tethered flies flying in the dark require JO neurons to generate slow antiphasic oscillation of the left and right wing stroke amplitudes. However, JO neurons are not necessary for this antiphasic oscillation when visual feedback is available, indicating that there are multiple pathways for generating antiphasic movement of the wings. Collectively, our results are consistent with a model in which flying flies use JO neurons to detect increases in the wing-induced airflow and that JO neurons are involved in a response that decreases contralateral wing stoke amplitude. PMID:25995481

  7. [First-tracer passage with a single-crystal gamma camera: completed assessment of left-ventricular function by determining enddiastolic volume, regional ejection fraction and %-akinetic segment (author's transl)].

    PubMed

    Bull, U; Knesewitsch, P; Kleinhans, E; Seiderer, M; Strauer, B E

    1981-06-01

    Determination of left ventricular (LV) enddiastolic volume (EDV) was achieved by calibration of the system (single-crystal gamma camera, equipped with a converging collimator) to a volume phantom (egg). A good correlation (r = 0.92) was found with EDV values, obtained from cineventriculography. Images, derived from enddiastole (ED) and endsystole (ES) were corrected for background by "parabolic background subtraction", which is a realistic form of background correction in view of the LV-shape. Regional ejection fraction (REF) was calculated by an electronical operation using the ejection fraction formula and these ED and ES images. REF values reflect regional or segmental LV pump function and are superior to one- or two-dimensional parameters (e.g. visual assessment of asynergy, hemiaxis shortening) since REF values include the third dimension by referring to regional volumes. In addition, per cent-akinetic segment may be replaced by REF. Results from the literature show that first-tracer passage with a single crystal gamma camera at rest (n = 534) yield equivalent results in comparison with cineventriculography. Therefore, this nuclear procedure may be routinely used. REF values complete the diagnostic parameter as yet available. PMID:6265871

  8. Plasma Metabolic Profile Determination in Young ST-segment Elevation Myocardial Infarction Patients with Ischemia and Reperfusion: Ultra-performance Liquid Chromatography and Mass Spectrometry for Pathway Analysis

    PubMed Central

    Huang, Lei; Li, Tong; Liu, Ying-Wu; Zhang, Lei; Dong, Zhi-Huan; Liu, Shu-Ye; Gao, Ying-Tang

    2016-01-01

    Background: This study was to establish a disease differentiation model for ST-segment elevation myocardial infarction (STEMI) youth patients experiencing ischemia and reperfusion via ultra-performance liquid chromatography and mass spectrometry (UPLC/MS) platform, which searches for closely related characteristic metabolites and metabolic pathways to evaluate their predictive value in the prognosis after discharge. Methods: Forty-seven consecutive STEMI patients (23 patients under 45 years of age, referred to here as “youth,” and 24 “elderly” patients) and 48 healthy control group members (24 youth, 24 elderly) were registered prospectively. The youth patients were required to provide a second blood draw during a follow-up visit one year after morbidity (n = 22, one lost). Characteristic metabolites and relative metabolic pathways were screened via UPLC/MS platform base on the Kyoto encyclopedia of genes and genomes (KEGG) and Human Metabolome Database. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of characteristic metabolites in the prognosis after discharge. Results: We successfully established an orthogonal partial least squares discriminated analysis model (R2X = 71.2%, R2Y = 79.6%, and Q2 = 55.9%) and screened out 24 ions; the sphingolipid metabolism pathway showed the most drastic change. The ROC curve analysis showed that ceramide [Cer(d18:0/16:0), Cer(t18:0/12:0)] and sphinganine in the sphingolipid pathway have high sensitivity and specificity on the prognosis related to major adverse cardiovascular events after youth patients were discharged. The area under curve (AUC) was 0.671, 0.750, and 0.711, respectively. A follow-up validation one year after morbidity showed corresponding AUC of 0.778, 0.833, and 0.806. Conclusions: By analyzing the plasma metabolism of myocardial infarction patients, we successfully established a model that can distinguish two different factors simultaneously: pathological

  9. Millisecond single-molecule localization microscopy combined with convolution analysis and automated image segmentation to determine protein concentrations in complexly structured, functional cells, one cell at a time.

    PubMed

    Wollman, Adam J M; Leake, Mark C

    2015-01-01

    We present a single-molecule tool called the CoPro (concentration of proteins) method that uses millisecond imaging with convolution analysis, automated image segmentation and super-resolution localization microscopy to generate robust estimates for protein concentration in different compartments of single living cells, validated using realistic simulations of complex multiple compartment cell types. We demonstrate its utility experimentally on model Escherichia coli bacteria and Saccharomyces cerevisiae budding yeast cells, and use it to address the biological question of how signals are transduced in cells. Cells in all domains of life dynamically sense their environment through signal transduction mechanisms, many involving gene regulation. The glucose sensing mechanism of S. cerevisiae is a model system for studying gene regulatory signal transduction. It uses the multi-copy expression inhibitor of the GAL gene family, Mig1, to repress unwanted genes in the presence of elevated extracellular glucose concentrations. We fluorescently labelled Mig1 molecules with green fluorescent protein (GFP) via chromosomal integration at physiological expression levels in living S. cerevisiae cells, in addition to the RNA polymerase protein Nrd1 with the fluorescent protein reporter mCherry. Using CoPro we make quantitative estimates of Mig1 and Nrd1 protein concentrations in the cytoplasm and nucleus compartments on a cell-by-cell basis under physiological conditions. These estimates indicate a ∼4-fold shift towards higher values in the concentration of diffusive Mig1 in the nucleus if the external glucose concentration is raised, whereas equivalent levels in the cytoplasm shift to smaller values with a relative change an order of magnitude smaller. This compares with Nrd1 which is not involved directly in glucose sensing, and which is almost exclusively localized in the nucleus under high and low external glucose levels. CoPro facilitates time-resolved quantification of

  10. The Dynamics in Epithelial Cell Intercalation in Drosophila Morphogenesis

    NASA Astrophysics Data System (ADS)

    Wolf, Fred; Reichl, Lars; Kong, Deqing; Zhang, Yujun; Eule, Stephan; Metzger, Jakob; Großhans, Jörg

    2015-03-01

    Epithelial cell rearrangement is important for many processes in morphogenesis. During germband extension in early gastrulation of Drosophila embryos, exchange of neighbors is achieved by junction remodeling that follows a topological T1 process. Its first step is the constriction of dorsal-ventral junctions and fusion of two 3x vertices into a 4x vertex a process believed to be junction autonomous. We established a high throughput imaging pipeline, by which we recorded, segmented and analysed more than 1000 neighbor exchanges in drosophila embryos. Characterizing the dynamics of junction lengths we find that the constriction of cell contacts follows intriguingly simple quantitative laws. (1) The mean contact length decreases approximately as a square root of time to collapse. (2) The time dependent variance of contact lengths is proportional to the square of the mean. (3) The time dependent probability density of the contact lengths remains close to Gaussian during the entire process. These observations are sufficient to derive a stochastic differential equation for contact length that captures the non-equilibrium statistical mechanics of contact collapse. Supported by the German Research Foundation.

  11. Genetic Architecture of Abdominal Pigmentation in Drosophila melanogaster

    PubMed Central

    Dembeck, Lauren M.; Huang, Wen; Magwire, Michael M.; Lawrence, Faye; Lyman, Richard F.; Mackay, Trudy F. C.

    2015-01-01

    Pigmentation varies within and between species and is often adaptive. The amount of pigmentation on the abdomen of Drosophila melanogaster is a relatively simple morphological trait, which serves as a model for mapping the genetic basis of variation in complex phenotypes. Here, we assessed natural variation in female abdominal pigmentation in 175 sequenced inbred lines of the Drosophila melanogaster Genetic Reference Panel, derived from the Raleigh, NC population. We quantified the proportion of melanization on the two most posterior abdominal segments, tergites 5 and 6 (T5, T6). We found significant genetic variation in the proportion of melanization and high broad-sense heritabilities for each tergite. Genome-wide association studies identified over 150 DNA variants associated with the proportion of melanization on T5 (84), T6 (34), and the difference between T5 and T6 (35). Several of the top variants associated with variation in pigmentation are in tan, ebony, and bric-a-brac1, genes known to affect D. melanogaster abdominal pigmentation. Mutational analyses and targeted RNAi-knockdown showed that 17 out of 28 (61%) novel candidate genes implicated by the genome-wide association study affected abdominal pigmentation. Several of these genes are involved in developmental and regulatory pathways, chitin production, cuticle structure, and vesicle formation and transport. These findings show that genetic variation may affect multiple steps in pathways involved in tergite development and melanization. Variation in these novel candidates may serve as targets for adaptive evolution and sexual selection in D. melanogaster. PMID:25933381

  12. The Homeotic Gene Sex Combs Reduced of Drosophila Melanogaster Is Differentially Regulated in the Embryonic and Imaginal Stages of Development

    PubMed Central

    Pattatucci, A. M.; Kaufman, T. C.

    1991-01-01

    The Sex combs reduced (Scr) locus is unique among the genes contained within the Antennapedia complex (ANT-C) of Drosophila melanogaster in that it directs functions that are required for both cephalic and thoracic development in the embryo and the adult. Antibodies raised against protein encoded by Scr were used to follow the distribution of this gene product in embryos and imaginal discs of third instar larvae. Analysis of Scr protein accumulation in embryos hemizygous for breakpoint lesions mapping throughout the locus has allowed us to determine that sequences required for establishment of the Scr embryonic pattern are contained within a region of DNA that overlaps with the identified upstream regulatory region of the segmentation gene fushi tarazu (ftz). Gain-of-function mutations in Scr result in the presence of ectopic sex comb teeth on the first tarsal segment of mesothoracic and metathoracic legs of adult males. Heterozygous combinations of gain-of-function alleles with a wild-type Scr gene exhibit no evidence of ectopic protein localization in the second and third thoracic segments of embryos. However, mesothoracic and metathoracic leg imaginal discs can be shown to accumulate ectopically expressed Scr protein, implying a differential regulation of the Scr gene during these two periods of development. Additionally, we have found that the spatial pattern of Scr gene expression in imaginal tissues involved in the development of the adult thorax is governed in part by synapsis of homologous chromosomes in this region of the ANT-C. However, those imaginal discs that arise anteriorly to the prothorax do not appear to be sensitive to this form of gene regulation. Finally, we have demonstrated that the extent of Scr expression is influenced by mutations at the Polycomb (Pc) locus but not by mutant alleles of the zeste (z) gene. Taken together, our data suggests that Scr gene expression is differentially regulated both temporally and spatially in a manner that is

  13. Genetic basis of the difference in alcohol dehydrogenase expression between Drosophila melanogaster and Drosophila simulans.

    PubMed Central

    Laurie, C C; Heath, E M; Jacobson, J W; Thomson, M S

    1990-01-01

    Drosophila melanogaster and its sibling species, Drosophila simulans, differ in expression of the enzyme alcohol dehydrogenase (ADH). Adult melanogaster flies that are homozygous for the Slow allozyme have approximately twice the level of ADH activity and crossreacting material as simulans adults. There is no corresponding difference in ADH mRNA, however, so this difference in ADH protein level is evidently due to a difference in the rate of translation of the two RNAs and/or to a difference in protein stability. Here we report an interspecific gene-transfer experiment, using P-element transformation, to determine whether this expression difference is due to genetic background differences between the species (trans-acting modifiers) or to cis-acting factors within the Adh gene. When the Adh genes from D. melanogaster and D. simulans are put into the same genetic background, there is no detectable difference in their level of expression. The level is relatively high in the melanogaster background and relatively low in the simulans background. Therefore, the interspecific difference in Adh expression is due entirely to trans-acting modifiers, in spite of the many sequence differences between the Adh genes of the two species, which include two amino acid substitutions. PMID:2124699

  14. Regulation of the Gene Sex-Lethal: A Comparative Analysis of Drosophila Melanogaster and Drosophila Subobscura

    PubMed Central

    Penalva, LOF.; Sakamoto, H.; Navarro-Sabate, A.; Sakashita, E.; Granadino, B.; Segarra, C.; Sanchez, L.

    1996-01-01

    The Drosophila gene Sex-lethal (Sxl) controls the processes of sex determination and dosage compensation. A Drosophila subobscura genomic fragment containing all the exons and the late and early promotors in the Sxl gene of D. melanogaster was isolated. Early Sxl expression in D. subobscura seems to be controlled at the transcriptional level, possibly by the X:A signal. In the region upstream of the early Sxl transcription initiation site are two conserved regions suggested to be involved in the early activation of Sxl. Late Sxl expression in D. subobscura produces four transcripts in adult females and males. In males, the transcripts have an additional exon which contains three translational stop codons so that a truncated, presumably nonfunctional Sxl protein is produced. The Sxl pre-mRNA of D. subobscura lacks the poly-U sequence presented at the polypirimidine tract of the 3' splice site of the male-specific exon present in D. melanogaster. Introns 2 and 3 contain the Sxl-binding poly-U stretches, whose localization in intron 2 varies but in intron 3 is conserved. The Sxl protein is fully conserved at the amino acid level in both species. PMID:8978052

  15. Vitrification-based cryopreservation of Drosophila embryos

    SciTech Connect

    Schreuders, P.D.; Mazur, P.

    1994-12-31

    Currently, over 30,000 strains of Drosophila melanogaster are maintained by geneticists through regular transfer of breeding stocks. A more cost effective solution is to cryopreserve their embryos. Cooling and warming rates >10,000{degrees}C/min. are required to prevent chilling injury. To avoid the lethal intracellular ice normally produced at such high cooling rates, it is necessary to use {ge}50% (w/w) concentrations of glass-inducing solutes to vitrify the embryos. Differential scanning calorimetry (DSC) is used to develop and evaluate ethylene glycol and polyvinyl pyrrolidone based vitrification solutions. The resulting solution consists of 8.5M ethylene glycol + 10% polyvinylpyrrolidone in D-20 Drosophila culture medium. A two stage method is used for the introduction and concentration of these solutes within the embryo. The method reduces the exposure time to the solution and, consequently, reduces toxicity. Both DSC and freezing experiments suggest that, while twelve-hour embryos will vitrify using cooling rates >200{degrees}C/min., they will devitrify and be killed with even moderately rapid warming rates of {approximately}1,900{degrees}C/min. Very rapid warming ({approximately}100,000{degrees}C/min.) results in variable numbers of successfully cryopreserved embryos. This sensitivity to warming rite is typical of devitrification. The variability in survival is reduced using embryos of a precisely determined embryonic stage. The vitrification of the older, fifteen-hour, embryos yields an optimized hatching rate of 68%, with 35 - 40% of the resulting larvae developing to normal adults. This Success rite in embryos of this age may reflect a reduced sensitivity to limited devitrification or a more even distribution of the ethylene glycol within the embryo.

  16. Structure and Development of Glia in Drosophila

    PubMed Central

    Hartenstein, Volker

    2014-01-01

    Insect glia represents a conspicuous and diverse population of cells and plays a role in controlling neuronal progenitor proliferation, axonal growth, neuronal differentiation and maintenance, and neuronal function. Genetic studies in Drosophila have elucidated many aspects of glial structure, function and development. Just as in vertebrates, it appears as if different classes of glial cells are specialized for different functions. Based on topology and cell shape, glial cells of the central nervous system fall into three classes (Fig. 1A–C): (i) surface glia that extend sheath-like processes to wrap around the entire brain; (ii) cortex glia (also called cell body-associated glia) that encapsulate neuronal somata and neuroblasts which form the outer layer (cortex) of the central nervous system; (iii) neuropile glia that are located at the interface between the cortex and the neuropile, the central domain of the nervous system formed by the highly branched neuronal processes and their synaptic contacts. Surface glia is further subdivided into an outer, perineurial layer, and an inner, subperineurial layer. Likewise, neuropile glia comprises a class of cells that remain at the surface of the neuropile (ensheathing glia), and a second class that forms profuse lamellar processes around nerve fibers within the neuropile (astrocyte-like or reticular glia). Glia also surrounds the peripheral nerves and sensory organs; here, one also recognizes perineurial and subperineurial glia, and a third type called “wrapping glia” that most likely corresponds to the ensheathing glia of the central nervous system. Much more experimental work is needed to determine how fundamental these differences between classes of glial cells are, or how and when during development they are specified. To aid in this work the following review will briefly summarize our knowledge of the classes of glial cells encountered in the Drosophila nervous system, and then survey their development from

  17. Drosophila Ten-m and Filamin Affect Motor Neuron Growth Cone Guidance

    PubMed Central

    Zheng, Lihua; Michelson, Yehudit; Freger, Vita; Avraham, Ziva; Venken, Koen J. T.; Bellen, Hugo J.; Justice, Monica J.; Wides, Ron

    2011-01-01

    The Drosophila Ten-m (also called Tenascin-major, or odd Oz (odz)) gene has been associated with a pair-rule phenotype. We identified and characterized new alleles of Drosophila Ten-m to establish that this gene is not responsible for segmentation defects but rather causes defects in motor neuron axon routing. In Ten-m mutants the inter-segmental nerve (ISN) often crosses segment boundaries and fasciculates with the ISN in the adjacent segment. Ten-m is expressed in the central nervous system and epidermal stripes during the stages when the growth cones of the neurons that form the ISN navigate to their targets. Over-expression of Ten-m in epidermal cells also leads to ISN misrouting. We also found that Filamin, an actin binding protein, physically interacts with the Ten-m protein. Mutations in cheerio, which encodes Filamin, cause defects in motor neuron axon routing like those of Ten-m. During embryonic development, the expression of Filamin and Ten-m partially overlap in ectodermal cells. These results suggest that Ten-m and Filamin in epidermal cells might together influence growth cone progression. PMID:21857973

  18. Native Microbial Colonization of Drosophila melanogaster and Its Use as a Model of Enterococcus faecalis Pathogenesis▿ †

    PubMed Central

    Cox, Christopher R.; Gilmore, Michael S.

    2007-01-01

    Enterococci are commensal organisms of the gastrointestinal (GI) tracts of a broad range of mammalian and insect hosts, but they are also leading causes of nosocomial infection. Little is known about the ecological role of enterococci in the GI tract consortia. To develop a tractable model for studying the roles of these organisms as commensals and pathogens, we characterized the Drosophila melanogaster microflora and examined the occurrence of enterococci in the gastrointestinal consortium of Drosophila. In a survey of laboratory-reared Drosophila and wild-captured flies, we found that Drosophila was naturally colonized by representatives of five bacterial phyla. Among these organisms were several species of enterococci, including Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinaraum, and Enterococcus durans, as well as a previously detected but uncultured Enterococcus species. Drosophila could be cured of enterococcal carriage by antibiotic treatment and could be reassociated with laboratory strains. High-level colonization by a well-characterized strain expressing the enterococcal cytolysin was found to be detrimental to Drosophila compared to the effect of an isogenic, noncytolytic control. The anatomical distribution of enterococci in the Drosophila GI tract was determined by immunohistochemical staining of thin sections of naturally colonized and reassociated flies. PMID:17220307

  19. Methodologies to determine forces on bones and muscles of body segments during exercise, employing compact sensors suitable for use in crowded space vehicles

    NASA Technical Reports Server (NTRS)

    Figueroa, Fernando

    1994-01-01

    A complete description of an instrumented ergometer system, including the sensors, the data acquisition system, and the methodologies to calculate the kinematic parameters were initially developed at Tulane University. This work was continued by the PI at NASA Johnson Space Center, where a flight ergometer was instrumented and tested during a KC-135 Zero-Gravity flight. The sensors that form part of the system include EMG probes and accelerometers mounted on the subject using the ergometer, load cells to measure pedal forces, and encoders to measure position and orientation of the pedal (foot). Currently, data from the flight test is being analyzed and processed to calculate the kinematic parameters of the individual. The formulation developed during the initial months of the grant will be used for this purpose. The system's components are compact (all sensors are very small). A salient feature of the system and associated methodology to determine the kinematics is that although it uses accelerometers, position is not determined by integration. Position is determined by determining the angle of two frames of reference for which acceleration at one point is known in coordinates of both frames.

  20. The evolving role of the orphan nuclear receptor ftz-f1, a pair-rule segmentation gene.

    PubMed

    Heffer, Alison; Grubbs, Nathaniel; Mahaffey, James; Pick, Leslie

    2013-01-01

    Segmentation is a critical developmental process that occurs by different mechanisms in diverse taxa. In insects, there are three common modes of embryogenesis-short-, intermediate-, and long-germ development-which differ in the number of segments specified at the blastoderm stage. While genes involved in segmentation have been extensively studied in the long-germ insect Drosophila melanogaster (Dm), it has been found that their expression and function in segmentation in short- and intermediate-germ insects often differ. Drosophila ftz-f1 encodes an orphan nuclear receptor that functions as a maternally expressed pair-rule segmentation gene, responsible for the formation of alternate body segments during Drosophila embryogenesis. Here we investigated the expression and function of ftz-f1 in the short-germ beetle, Tribolium castaneum (Tc). We found that Tc-ftz-f1 is expressed in stripes in Tribolium embryos. These stripes overlap alternate Tc-Engrailed (Tc-En) stripes, indicative of a pair-rule expression pattern. To test whether Tc-ftz-f1 has pair-rule function, we utilized embryonic RNAi, injecting double-stranded RNA corresponding to Tc-ftz-f1 coding or non-coding regions into early Tribolium embryos. Knockdown of Tc-ftz-f1 produced pair-rule segmentation defects, evidenced by loss of expression of alternate En stripes. In addition, a later role for Tc-ftz-f1 in cuticle formation was revealed. These results identify a new pair-rule gene in Tribolium and suggest that its role in segmentation may be shared among holometabolous insects. Interestingly, while Tc-ftz-f1 is expressed in pair-rule stripes, the gene is ubiquitously expressed in Drosophila embryos. Thus, the pair-rule function of ftz-f1 is conserved despite differences in expression patterns of ftz-f1 genes in different lineages. This suggests that ftz-f1 expression changed after the divergence of lineages leading to extant beetles and flies, likely due to differences in cis-regulatory sequences. We

  1. Pancreas and cyst segmentation

    NASA Astrophysics Data System (ADS)

    Dmitriev, Konstantin; Gutenko, Ievgeniia; Nadeem, Saad; Kaufman, Arie

    2016-03-01

    Accurate segmentation of abdominal organs from medical images is an essential part of surgical planning and computer-aided disease diagnosis. Many existing algorithms are specialized for the segmentation of healthy organs. Cystic pancreas segmentation is especially challenging due to its low contrast boundaries, variability in shape, location and the stage of the pancreatic cancer. We present a semi-automatic segmentation algorithm for pancreata with cysts. In contrast to existing automatic segmentation approaches for healthy pancreas segmentation which are amenable to atlas/statistical shape approaches, a pancreas with cysts can have even higher variability with respect to the shape of the pancreas due to the size and shape of the cyst(s). Hence, fine results are better attained with semi-automatic steerable approaches. We use a novel combination of random walker and region growing approaches to delineate the boundaries of the pancreas and cysts with respective best Dice coefficients of 85.1% and 86.7%, and respective best volumetric overlap errors of 26.0% and 23.5%. Results show that the proposed algorithm for pancreas and pancreatic cyst segmentation is accurate and stable.

  2. Determinant for β-subunit regulation in high-conductance voltage-activated and Ca2+-sensitive K+ channels: An additional transmembrane region at the N terminus

    PubMed Central

    Wallner, Martin; Meera, Pratap; Toro, Ligia

    1996-01-01

    The pore-forming α subunit of large conductance voltage- and Ca2+-sensitive K (MaxiK) channels is regulated by a β subunit that has two membrane-spanning regions separated by an extracellular loop. To investigate the structural determinants in the pore-forming α subunit necessary for β-subunit modulation, we made chimeric constructs between a human MaxiK channel and the Drosophila homologue, which we show is insensitive to β-subunit modulation, and analyzed the topology of the α subunit. A comparison of multiple sequence alignments with hydrophobicity plots revealed that MaxiK channel α subunits have a unique hydrophobic segment (S0) at the N terminus. This segment is in addition to the six putative transmembrane segments (S1–S6) usually found in voltage-dependent ion channels. The transmembrane nature of this unique S0 region was demonstrated by in vitro translation experiments. Moreover, normal functional expression of signal sequence fusions and in vitro N-linked glycosylation experiments indicate that S0 leads to an exoplasmic N terminus. Therefore, we propose a new model where MaxiK channels have a seventh transmembrane segment at the N terminus (S0). Chimeric exchange of 41 N-terminal amino acids, including S0, from the human MaxiK channel to the Drosophila homologue transfers β-subunit regulation to the otherwise unresponsive Drosophila channel. Both the unique S0 region and the exoplasmic N terminus are necessary for this gain of function. PMID:8962157

  3. Determinant for beta-subunit regulation in high-conductance voltage-activated and Ca(2+)-sensitive K+ channels: an additional transmembrane region at the N terminus.

    PubMed

    Wallner, M; Meera, P; Toro, L

    1996-12-10

    The pore-forming alpha subunit of large conductance voltage- and Ca(2+)-sensitive K (MaxiK) channels is regulated by a beta subunit that has two membrane-spanning regions separated by an extracellular loop. To investigate the structural determinants in the pore-forming alpha subunit necessary for beta-subunit modulation, we made chimeric constructs between a human MaxiK channel and the Drosophila homologue, which we show is insensitive to beta-subunit modulation, and analyzed the topology of the alpha subunit. A comparison of multiple sequence alignments with hydrophobicity plots revealed that MaxiK channel alpha subunits have a unique hydrophobic segment (S0) at the N terminus. This segment is in addition to the six putative transmembrane segments (S1-S6) usually found in voltage-dependent ion channels. The transmembrane nature of this unique S0 region was demonstrated by in vitro translation experiments. Moreover, normal functional expression of signal sequence fusions and in vitro N-linked glycosylation experiments indicate that S0 leads to an exoplasmic N terminus. Therefore, we propose a new model where MaxiK channels have a seventh transmembrane segment at the N terminus (S0). Chimeric exchange of 41 N-terminal amino acids, including S0, from the human MaxiK channel to the Drosophila homologue transfers beta-subunit regulation to the otherwise unresponsive Drosophila channel. Both the unique S0 region and the exoplasmic N terminus are necessary for this gain of function. PMID:8962157

  4. cAMP signalling in mushroom bodies modulates temperature preference behaviour in Drosophila.

    PubMed

    Hong, Sung-Tae; Bang, Sunhoe; Hyun, Seogang; Kang, Jongkyun; Jeong, Kyunghwa; Paik, Donggi; Chung, Jongkyeong; Kim, Jaeseob

    2008-08-01

    Homoiotherms, for example mammals, regulate their body temperature with physiological responses such as a change of metabolic rate and sweating. In contrast, the body temperature of poikilotherms, for example Drosophila, is the result of heat exchange with the surrounding environment as a result of the large ratio of surface area to volume of their bodies. Accordingly, these animals must instinctively move to places with an environmental temperature as close as possible to their genetically determined desired temperature. The temperature that Drosophila instinctively prefers has a function equivalent to the 'set point' temperature in mammals. Although various temperature-gated TRP channels have been discovered, molecular and cellular components in Drosophila brain responsible for determining the desired temperature remain unknown. We identified these components by performing a large-scale genetic screen of temperature preference behaviour (TPB) in Drosophila. In parallel, we mapped areas of the Drosophila brain controlling TPB by targeted inactivation of neurons with tetanus toxin and a potassium channel (Kir2.1) driven with various brain-specific GAL4s. Here we show that mushroom bodies (MBs) and the cyclic AMP-cAMP-dependent protein kinase A (cAMP-PKA) pathway are essential for controlling TPB. Furthermore, targeted expression of cAMP-PKA pathway components in only the MB was sufficient to rescue abnormal TPB of the corresponding mutants. Preferred temperatures were affected by the level of cAMP and PKA activity in the MBs in various PKA pathway mutants. PMID:18594510

  5. Distinct Biochemical Activities of Eyes absent During Drosophila Eye Development

    PubMed Central

    Jin, Meng; Mardon, Graeme

    2016-01-01

    Eyes absent (Eya) is a highly conserved transcriptional coactivator and protein phosphatase that plays vital roles in multiple developmental processes from Drosophila to humans. Eya proteins contain a PST (Proline-Serine-Threonine)-rich transactivation domain, a threonine phosphatase motif (TPM), and a tyrosine protein phosphatase domain. Using a genomic rescue system, we find that the PST domain is essential for Eya activity and Dac expression, and the TPM is required for full Eya function. We also find that the threonine phosphatase activity plays only a minor role during Drosophila eye development and the primary function of the PST and TPM domains is transactivation that can be largely substituted by the heterologous activation domain VP16. Along with our previous results that the tyrosine phosphatase activity of Eya is dispensable for normal Eya function in eye formation, we demonstrate that a primary function of Eya during Drosophila eye development is as a transcriptional coactivator. Moreover, the PST/TPM and the threonine phosphatase activity are not required for in vitro interaction between retinal determination factors. Finally, this work is the first report of an Eya-Ey physical interaction. These findings are particularly important because they highlight the need for an in vivo approach that accurately dissects protein function. PMID:26980695

  6. Evaluation of polylactic acid nanoparticles safety using Drosophila model.

    PubMed

    Legaz, Sophie; Exposito, Jean-Yves; Lethias, Claire; Viginier, Barbara; Terzian, Christophe; Verrier, Bernard

    2016-10-01

    Cytotoxicity of nanoparticles and their sub-lethal effect on cell behavior and cell fate are a high topic of studies in the nanomaterial field. With an explosion of nanoparticle types (size, shape, polarity, stiffness, composition, etc.), Drosophila has become an attractive animal model for high throughput analysis of these nanocarriers in the drug delivery field with applications in cancer therapy, or simply to generate a fast and complete cytotoxic study of a peculiar nanoparticle. In respect to that, we have conducted an in cellulo study of poly(lactic acid) (PLA) nanoparticle cytotoxicity, and determined that near lethal nanoparticle doses, oxidative stress as well as P53 and ATP pathways may lead to cell cycle arrest at G1, and ultimately to cell death. Neither viability nor the development of Drosophila larvae are affected by the ingestion of PLA nanoparticles at sub-lethal concentrations. Drosophila will be a useful model to study PLA and PLA-modified nanoparticle toxicity, and nanoparticle fate after ingestion. PMID:27108761

  7. Distinct Biochemical Activities of Eyes absent During Drosophila Eye Development.

    PubMed

    Jin, Meng; Mardon, Graeme

    2016-01-01

    Eyes absent (Eya) is a highly conserved transcriptional coactivator and protein phosphatase that plays vital roles in multiple developmental processes from Drosophila to humans. Eya proteins contain a PST (Proline-Serine-Threonine)-rich transactivation domain, a threonine phosphatase motif (TPM), and a tyrosine protein phosphatase domain. Using a genomic rescue system, we find that the PST domain is essential for Eya activity and Dac expression, and the TPM is required for full Eya function. We also find that the threonine phosphatase activity plays only a minor role during Drosophila eye development and the primary function of the PST and TPM domains is transactivation that can be largely substituted by the heterologous activation domain VP16. Along with our previous results that the tyrosine phosphatase activity of Eya is dispensable for normal Eya function in eye formation, we demonstrate that a primary function of Eya during Drosophila eye development is as a transcriptional coactivator. Moreover, the PST/TPM and the threonine phosphatase activity are not required for in vitro interaction between retinal determination factors. Finally, this work is the first report of an Eya-Ey physical interaction. These findings are particularly important because they highlight the need for an in vivo approach that accurately dissects protein function. PMID:26980695

  8. Parasitoid virus-like particles destroy Drosophila cellular immunity.

    PubMed

    Rizki, R M; Rizki, T M

    1990-11-01

    Parasitoid wasps must avoid the destructive effects of the host's cellular defense system in order to exploit the host hemocoel as a suitable environment for their survival. To protect their eggs from encapsulation by Drosophila melanogaster blood cells, Leptopilina heterotoma females inject a factor that selectively destroys lamellocytes, the type of Drosophila blood cell involved in recognition and encapsulation of large foreign objects. Other types of host blood cells, including the phagocytic plasmatocytes, remain functional. This report demonstrates that the destructive factor for lamellocytes is a virus-like particle (VLP) stored in the reservoir of an accessory gland associated with the female wasp reproductive system. We show that VLPs enter Drosophila blood cells in vitro. VLPs are found free in the cytoplasm of lamellocytes but are confined to phagocytic vesicles of plasmatocytes. As lamellocytes are susceptible to the VLP infection and plasmatocytes are not, we conclude that the mode of VLP entry and its disposition in the cytoplasm determine the fate of the infected host blood cell. PMID:2122461

  9. Design and Generation of Drosophila Single Guide RNA Expression Constructs.

    PubMed

    Housden, Benjamin E; Hu, Yanhui; Perrimon, Norbert

    2016-01-01

    The recent advances in CRISPR-based genome engineering have enabled a plethora of new experiments to study a wide range of biological questions. The major attraction of this system over previous methods is its high efficiency and simplicity of use. For example, whereas previous genome engineering technologies required the generation of new proteins to target each new locus, CRISPR requires only the expression of a different single guide RNA (sgRNA). This sgRNA binds to the Cas9 endonuclease protein and directs the generation of a double-strand break to a highly specific genomic site determined by the sgRNA sequence. In addition, the relative simplicity of the Drosophila genome is a particular advantage, as possible sgRNA off-target sites can easily be avoided. Here, we provide a step-by-step protocol for designing sgRNA target sites using the Drosophila RNAi Screening Center (DRSC) Find CRISPRs tool (version 2). We also describe the generation of sgRNA expression plasmids for the use in cultured Drosophila cells or in vivo. Finally, we discuss specific design requirements for various genome engineering applications. PMID:27587779

  10. Gut microbiota dictates the metabolic response of Drosophila to diet

    PubMed Central

    Wong, Adam C.-N.; Dobson, Adam J.; Douglas, Angela E.

    2014-01-01

    Animal nutrition is profoundly influenced by the gut microbiota, but knowledge of the scope and core mechanisms of the underlying animal–microbiota interactions is fragmentary. To investigate the nutritional traits shaped by the gut microbiota of Drosophila, we determined the microbiota-dependent response of multiple metabolic and performance indices to systematically varied diet composition. Diet-dependent differences between Drosophila bearing its unmanipulated microbiota (conventional flies) and experimentally deprived of its microbiota (axenic flies) revealed evidence for: microbial sparing of dietary B vitamins, especially riboflavin, on low-yeast diets; microbial promotion of protein nutrition, particularly in females; and microbiota-mediated suppression of lipid/carbohydrate storage, especially on high sugar diets. The microbiota also sets the relationship between energy storage and body mass, indicative of microbial modulation of the host signaling networks that coordinate metabolism with body size. This analysis identifies the multiple impacts of the microbiota on the metabolism of Drosophila, and demonstrates that the significance of these different interactions varies with diet composition and host sex. PMID:24577449

  11. A drosophila full-length cDNA resource

    SciTech Connect

    Stapleton, Mark; Carlson, Joseph; Brokstein, Peter; Yu, Charles; Champe, Mark; George, Reed; Guarin, Hannibal; Kronmiller, Brent; Pacleb, Joanne; Park, Soo; Rubin, Gerald M.; Celniker, Susan E.

    2003-05-09

    Background: A collection of sequenced full-length cDNAs is an important resource both for functional genomics studies and for the determination of the intron-exon structure of genes. Providing this resource to the Drosophila melanogaster research community has been a long-term goal of the Berkeley Drosophila Genome Project. We have previously described the Drosophila Gene Collection (DGC), a set of putative full-length cDNAs that was produced by generating and analyzing over 250,000 expressed sequence tags (ESTs) derived from a variety of tissues and developmental stages. Results: We have generated high-quality full-insert sequence for 8,921 clones in the DGC. We compared the sequence of these clones to the annotated Release 3 genomic sequence, and identified more than 5,300 cDNAs that contain a complete and accurate protein-coding sequence. This corresponds to at least one splice form for 40 percent of the predicted D. melanogaster genes. We also identified potential new cases of RNA editing. Conclusions: We show that comparison of cDNA sequences to a high-quality annotated genomic sequence is an effective approach to identifying and eliminating defective clones from a cDNA collection and ensure its utility for experimentation. Clones were eliminated either because they carry single nucleotide discrepancies, which most probably result from reverse transcriptase errors, or because they are truncated and contain only part of the protein-coding sequence.

  12. Automatic Contrail Detection and Segmentation

    NASA Technical Reports Server (NTRS)

    Weiss, John M.; Christopher, Sundar A.; Welch, Ronald M.

    1998-01-01

    Automatic contrail detection is of major importance in the study of the atmospheric effects of aviation. Due to the large volume of satellite imagery, selecting contrail images for study by hand is impractical and highly subject to human error. It is far better to have a system in place that will automatically evaluate an image to determine 1) whether it contains contrails and 2) where the contrails are located. Preliminary studies indicate that it is possible to automatically detect and locate contrails in Advanced Very High Resolution Radiometer (AVHRR) imagery with a high degree of confidence. Once contrails have been identified and localized in a satellite image, it is useful to segment the image into contrail versus noncontrail pixels. The ability to partition image pixels makes it possible to determine the optical properties of contrails, including optical thickness and particle size. In this paper, we describe a new technique for segmenting satellite images containing contrails. This method has good potential for creating a contrail climatology in an automated fashion. The majority of contrails are detected, rejecting clutter in the image, even cirrus streaks. Long, thin contrails are most easily detected. However, some contrails may be missed because they are curved, diffused over a large area, or present in short segments. Contrails average 2-3 km in width for the cases studied.

  13. Segmented ion thruster

    NASA Technical Reports Server (NTRS)

    Brophy, John R. (Inventor)

    1993-01-01

    Apparatus and methods for large-area, high-power ion engines comprise dividing a single engine into a combination of smaller discharge chambers (or segments) configured to operate as a single large-area engine. This segmented ion thruster (SIT) approach enables the development of 100-kW class argon ion engines for operation at a specific impulse of 10,000 s. A combination of six 30-cm diameter ion chambers operating as a single engine can process over 100 kW. Such a segmented ion engine can be operated from a single power processor unit.

  14. The Pingding segment of the Altyn Tagh Fault (91 °E): Holocene slip-rate determination from cosmogenic radionuclide dating of offset fluvial terraces

    DOE PAGESBeta

    Meriaux, A. -S.; Van der Woerd, J.; Tapponnier, P.; Ryerson, F. J.; Finkel, R. C.; Lasserre, C.; Xu, X.

    2012-09-25

    Morphochronologic slip-rates on the Altyn Tagh Fault (ATF) along the southern front of the Pingding Shan at ~90.5°E are determined by cosmogenic radionuclide (CRN) dating of seven offset terraces at two sites. The terraces are defined based upon morphology, elevation and dating, together with fieldwork and high-resolution satellite analysis. The majority of the CRN model ages fall within narrow ranges (<2 ka) on the four main terraces (T1, T2, T3 and T3′), and allow a detailed terrace chronology. Bounds on the terrace ages and offsets of 5 independent terraces yield consistent slip-rate estimates. The long-term slip-rate of 13.9 ± 1.1more » mm/yr is defined at the 95% confidence level, as the joint rate probability distribution of the rate derived from each independent terrace. It falls within the bounds of all the rates defined on the central Altyn Tagh Fault between the Cherchen He (86.4°E) and Akato Tagh (~88°E) sites. This rate is ~10 mm/yr less than the upper rate determined near Tura at ~87°E, in keeping with the inference of an eastward decreasing rate due to progressive loss of slip to thrusts branching off the fault southwards but it is greater than the 9 ± 4 mm/yr rate determined at ~90°E by GPS surveys and other geodetic short-term rates defined elsewhere along the ATF. Furthermore, whether such disparate rates will ultimately be reconciled by a better understanding of fault mechanics, resolved transient deformations during the seismic cycle or by more accurate measurements made with either approach remains an important issue.« less

  15. The Pingding segment of the Altyn Tagh Fault (91 °E): Holocene slip-rate determination from cosmogenic radionuclide dating of offset fluvial terraces

    SciTech Connect

    Meriaux, A. -S.; Van der Woerd, J.; Tapponnier, P.; Ryerson, F. J.; Finkel, R. C.; Lasserre, C.; Xu, X.

    2012-09-25

    Morphochronologic slip-rates on the Altyn Tagh Fault (ATF) along the southern front of the Pingding Shan at ~90.5°E are determined by cosmogenic radionuclide (CRN) dating of seven offset terraces at two sites. The terraces are defined based upon morphology, elevation and dating, together with fieldwork and high-resolution satellite analysis. The majority of the CRN model ages fall within narrow ranges (<2 ka) on the four main terraces (T1, T2, T3 and T3′), and allow a detailed terrace chronology. Bounds on the terrace ages and offsets of 5 independent terraces yield consistent slip-rate estimates. The long-term slip-rate of 13.9 ± 1.1 mm/yr is defined at the 95% confidence level, as the joint rate probability distribution of the rate derived from each independent terrace. It falls within the bounds of all the rates defined on the central Altyn Tagh Fault between the Cherchen He (86.4°E) and Akato Tagh (~88°E) sites. This rate is ~10 mm/yr less than the upper rate determined near Tura at ~87°E, in keeping with the inference of an eastward decreasing rate due to progressive loss of slip to thrusts branching off the fault southwards but it is greater than the 9 ± 4 mm/yr rate determined at ~90°E by GPS surveys and other geodetic short-term rates defined elsewhere along the ATF. Furthermore, whether such disparate rates will ultimately be reconciled by a better understanding of fault mechanics, resolved transient deformations during the seismic cycle or by more accurate measurements made with either approach remains an important issue.

  16. Determining recreational, scenic, and historical-cultural potentials of landscape features along a segment of the ancient Silk Road using factor analyzing.

    PubMed

    Kurdoglu, Oguz; Kurdoglu, Banu Cicek

    2010-11-01

    Generally, roads are structures connecting areas of special importance to each other (e.g., from urban areas to villages). In addition to functioning as transportation mediums, roads have aesthetic qualities by means of their recreational, scenic, and cultural functions. One of these unexplored roads, the Trabzon-Gümüşhane old state highway in the Northeast Turkey, the subject of this study, is quite unique for being a part of the historical Silk Road. The aim of the study is to determine the recreational, scenic, and cultural potentials of this road and to identify the landscape qualities based on these potentials. After dividing the road into three sections for comparison purposes, data gathered from basic digital maps obtained from The General Directorate of Rural Services (the governmental agency that is responsible for producing such digital maps) and field trips were entered into maps using geographic information system followed by creating demonstration films using photographs and video camera images of the natural and cultural characteristics of the area. Then, these films were evaluated by an expert team to identify the road's recreational, scenic, and cultural potentials. The results of the evaluation process, the relationships between determined landscape characteristics for each function, and the contribution scores of different functions assigned for the artificial features were statistically analyzed. The analyses showed a very strong relationship between the sections and the contribution scores of the recreation and the scenic functions. The results of these analyses also revealed some landscape characteristics having high contribution values for all the functions. This, in turn, helped in determining a potential of the study area to become a greenway. PMID:19862635

  17. Cryobiological preservation of Drosophila embryos

    SciTech Connect

    Mazur, P.; Schreuders, P.D.; Cole, K.W.; Hall, J.W. ); Mahowald, A.P. )

    1992-12-18

    The inability to cryobiologically preserve the fruit fly Drosophila melanogaster has required that fly stocks be maintained by frequent transfer of adults. This method is costly in terms of time and can lead to loss of stocks. Traditional slow freezing methods do not succeed because the embryos are highly sensitive to chilling. With the procedures described here, 68 percent of precisely staged 15-hour Oregon R (wild-type) embryos hatch after vitrification at -205[degree]C, and 40 percent of the resulting larvae develop into normal adult flies. These embryos are among the most complex organisms successfully preserved by cryobiology.

  18. Taste processing in Drosophila larvae

    PubMed Central

    Apostolopoulou, Anthi A.; Rist, Anna; Thum, Andreas S.

    2015-01-01

    The sense of taste allows animals to detect chemical substances in their environment to initiate appropriate behaviors: to find food or a mate, to avoid hostile environments and predators. Drosophila larvae are a promising model organism to study gustation. Their simple nervous system triggers stereotypic behavioral responses, and the coding of taste can be studied by genetic tools at the single cell level. This review briefly summarizes recent progress on how taste information is sensed and processed by larval cephalic and pharyngeal sense organs. The focus lies on several studies, which revealed cellular and molecular mechanisms required to process sugar, salt, and bitter substances. PMID:26528147

  19. Geotaxis baseline data for Drosophila

    NASA Technical Reports Server (NTRS)

    Schnebel, E. M.; Bhargava, R.; Grossfield, J.

    1987-01-01

    Geotaxis profiles for 20 Drosophila species and semispecies at different ages have been examined using a calibrated, adjustable slant board device. Measurements were taken at 5 deg intervals ranging from 0 deg to 85 deg. Clear strain and species differences are observed, with some groups tending to move upward (- geotaxis) with increasing angles, while others move downward (+ geotaxis). Geotactic responses change with age in some, but not all experimental groups. Sample geotaxis profiles are presented and their application to ecological and aging studies are discussed. Data provide a baseline for future evaluations of the biological effects of microgravity.

  20. Macrophages and cellular immunity in Drosophila melanogaster.

    PubMed

    Gold, Katrina S; Brückner, Katja

    2015-12-01

    The invertebrate Drosophila melanogaster has been a powerful model for understanding blood cell development and immunity. Drosophila is a holometabolous insect, which transitions through a series of life stages from embryo, larva and pupa to adulthood. In spite of this, remarkable parallels exist between Drosophila and vertebrate macrophages, both in terms of development and function. More than 90% of Drosophila blood cells (hemocytes) are macrophages (plasmatocytes), making this highly tractable genetic system attractive for studying a variety of questions in macrophage biology. In vertebrates, recent findings revealed that macrophages have two independent origins: self-renewing macrophages, which reside and proliferate in local microenvironments in a variety of tissues, and macrophages of the monocyte lineage, which derive from hematopoietic stem or progenitor cells. Like vertebrates, Drosophila possesses two macrophage lineages with a conserved dual ontogeny. These parallels allow us to take advantage of the Drosophila model when investigating macrophage lineage specification, maintenance and amplification, and the induction of macrophages and their progenitors by local microenvironments and systemic cues. Beyond macrophage development, Drosophila further serves as a paradigm for understanding the mechanisms underlying macrophage function and cellular immunity in infection, tissue homeostasis and cancer, throughout development and adult life. PMID:27117654

  1. Anatomical and Molecular Design of the Drosophila Antenna as a Flagellar Auditory Organ

    PubMed Central

    TODI, SOKOL V.; SHARMA, YASHODA; EBERL, DANIEL F.

    2007-01-01

    The molecular basis of hearing is less well understood than many other senses. However, recent studies in Drosophila have provided some important steps towards a molecular understanding of hearing. In this report, we summarize these findings and their implications on the relationship between hearing and touch. In Drosophila, hearing is accomplished by Johnston’s Organ, a chordotonal organ containing over 150 scolopidia within the second antennal segment. We will discuss anatomical features of the antenna and how they contribute to the function of this flagellar auditory receptor. The effects of several mutants, identified through mutagenesis screens or as homologues of vertebrate auditory genes, will be summarized. Based on evidence gathered from these studies, we propose a speculative model for how the chordotonal organ might function. PMID:15252880

  2. Selection Components in Background Replacement Lines of Drosophila

    PubMed Central

    Clark, Andrew G.; Bundgaard, Jørgen

    1984-01-01

    Selection components analysis was performed in lines of Drosophila melanogaster at three times during substitution backcrossing. The initial two lines were inbred isofemale lines from natural populations in California, and one had the spread wing mutation eagle. The selection components analysis revealed aspects of the genetic structure of the determinants of fitness by demonstrating changes in the marginal fitnesses of the eagle locus. Differences among backgrounds essentially disappeared by the 20th generation of backcrossing, suggesting that the previously observed differences were attributable to linkage disequilibrium. The method of bootstrapping was used as a novel means of determining statistical confidence in selection components. PMID:17246226

  3. Fitness and density-dependent population growth in Drosophila melanogaster

    SciTech Connect

    Mueller, L.D.; Ayala, F.J.

    1981-03-01

    The density-dependent rates of population growth were determined for 26 populations of Drosophila melanogaster maintained in the serial transfer system. Twenty-five populations were homozygous for an entire chromosome 2 sampled from nature; the other was a random heterozygous population. Rates of population growth around the carrying capacity cannot explain the large fitness depression of these lines. However, the homozygous lines show large differences in rates of population growth at low densities relative to the random heterozygous standard. The average relative fitness of the homozygous lines, as determined from the growth rates at the lowest density, is 0.51.

  4. Image segmentation survey

    NASA Technical Reports Server (NTRS)

    Haralick, R. M.

    1982-01-01

    The methodologies and capabilities of image segmentation techniques are reviewed. Single linkage schemes, hybrid linkage schemes, centroid linkage schemes, histogram mode seeking, spatial clustering, and split and merge schemes are addressed.

  5. Segmentation of SAR images

    NASA Technical Reports Server (NTRS)

    Kwok, Ronald

    1989-01-01

    The statistical characteristics of image speckle are reviewed. Existing segmentation techniques that have been used for speckle filtering, edge detection, and texture extraction are sumamrized. The relative effectiveness of each technique is briefly discussed.

  6. Neural circuits for peristaltic wave propagation in crawling Drosophila larvae: analysis and modeling

    PubMed Central

    Gjorgjieva, Julijana; Berni, Jimena; Evers, Jan Felix; Eglen, Stephen J.

    2013-01-01

    Drosophila larvae crawl by peristaltic waves of muscle contractions, which propagate along the animal body and involve the simultaneous contraction of the left and right side of each segment. Coordinated propagation of contraction does not require sensory input, suggesting that movement is generated by a central pattern generator (CPG). We characterized crawling behavior of newly hatched Drosophila larvae by quantifying timing and duration of segmental boundary contractions. We developed a CPG network model that recapitulates these patterns based on segmentally repeated units of excitatory and inhibitory (EI) neuronal populations coupled with immediate neighboring segments. A single network with symmetric coupling between neighboring segments succeeded in generating both forward and backward propagation of activity. The CPG network was robust to changes in amplitude and variability of connectivity strength. Introducing sensory feedback via “stretch-sensitive” neurons improved wave propagation properties such as speed of propagation and segmental contraction duration as observed experimentally. Sensory feedback also restored propagating activity patterns when an inappropriately tuned CPG network failed to generate waves. Finally, in a two-sided CPG model we demonstrated that two types of connectivity could synchronize the activity of two independent networks: connections from excitatory neurons on one side to excitatory contralateral neurons (E to E), and connections from inhibitory neurons on one side to excitatory contralateral neurons (I to E). To our knowledge, such I to E connectivity has not yet been found in any experimental system; however, it provides the most robust mechanism to synchronize activity between contralateral CPGs in our model. Our model provides a general framework for studying the conditions under which a single locally coupled network generates bilaterally synchronized and longitudinally propagating waves in either direction. PMID

  7. Neural circuits for peristaltic wave propagation in crawling Drosophila larvae: analysis and modeling.

    PubMed

    Gjorgjieva, Julijana; Berni, Jimena; Evers, Jan Felix; Eglen, Stephen J

    2013-01-01

    Drosophila larvae crawl by peristaltic waves of muscle contractions, which propagate along the animal body and involve the simultaneous contraction of the left and right side of each segment. Coordinated propagation of contraction does not require sensory input, suggesting that movement is generated by a central pattern generator (CPG). We characterized crawling behavior of newly hatched Drosophila larvae by quantifying timing and duration of segmental boundary contractions. We developed a CPG network model that recapitulates these patterns based on segmentally repeated units of excitatory and inhibitory (EI) neuronal populations coupled with immediate neighboring segments. A single network with symmetric coupling between neighboring segments succeeded in generating both forward and backward propagation of activity. The CPG network was robust to changes in amplitude and variability of connectivity strength. Introducing sensory feedback via "stretch-sensitive" neurons improved wave propagation properties such as speed of propagation and segmental contraction duration as observed experimentally. Sensory feedback also restored propagating activity patterns when an inappropriately tuned CPG network failed to generate waves. Finally, in a two-sided CPG model we demonstrated that two types of connectivity could synchronize the activity of two independent networks: connections from excitatory neurons on one side to excitatory contralateral neurons (E to E), and connections from inhibitory neurons on one side to excitatory contralateral neurons (I to E). To our knowledge, such I to E connectivity has not yet been found in any experimental system; however, it provides the most robust mechanism to synchronize activity between contralateral CPGs in our model. Our model provides a general framework for studying the conditions under which a single locally coupled network generates bilaterally synchronized and longitudinally propagating waves in either direction. PMID:23576980

  8. Adjacent segment disease.

    PubMed

    Virk, Sohrab S; Niedermeier, Steven; Yu, Elizabeth; Khan, Safdar N

    2014-08-01

    EDUCATIONAL OBJECTIVES As a result of reading this article, physicians should be able to: 1. Understand the forces that predispose adjacent cervical segments to degeneration. 2. Understand the challenges of radiographic evaluation in the diagnosis of cervical and lumbar adjacent segment disease. 3. Describe the changes in biomechanical forces applied to adjacent segments of lumbar vertebrae with fusion. 4. Know the risk factors for adjacent segment disease in spinal fusion. Adjacent segment disease (ASD) is a broad term encompassing many complications of spinal fusion, including listhesis, instability, herniated nucleus pulposus, stenosis, hypertrophic facet arthritis, scoliosis, and vertebral compression fracture. The area of the cervical spine where most fusions occur (C3-C7) is adjacent to a highly mobile upper cervical region, and this contributes to the biomechanical stress put on the adjacent cervical segments postfusion. Studies have shown that after fusion surgery, there is increased load on adjacent segments. Definitive treatment of ASD is a topic of continuing research, but in general, treatment choices are dictated by patient age and degree of debilitation. Investigators have also studied the risk factors associated with spinal fusion that may predispose certain patients to ASD postfusion, and these data are invaluable for properly counseling patients considering spinal fusion surgery. Biomechanical studies have confirmed the added stress on adjacent segments in the cervical and lumbar spine. The diagnosis of cervical ASD is complicated given the imprecise correlation of radiographic and clinical findings. Although radiological and clinical diagnoses do not always correlate, radiographs and clinical examination dictate how a patient with prolonged pain is treated. Options for both cervical and lumbar spine ASD include fusion and/or decompression. Current studies are encouraging regarding the adoption of arthroplasty in spinal surgery, but more long

  9. Segmented pyroelector detector

    DOEpatents

    Stotlar, S.C.; McLellan, E.J.

    1981-01-21

    A pyroelectric detector is described which has increased voltage output and improved responsivity over equivalent size detectors. The device comprises a plurality of edge-type pyroelectric detectors which have a length which is much greater than the width of the segments between the edge-type electrodes. External circuitry connects the pyroelectric detector segments in parallel to provide a single output which maintains 50 ohm impedance characteristics.

  10. Squaring a Circular Segment

    ERIC Educational Resources Information Center

    Gordon, Russell

    2008-01-01

    Consider a circular segment (the smaller portion of a circle cut off by one of its chords) with chord length c and height h (the greatest distance from a point on the arc of the circle to the chord). Is there a simple formula involving c and h that can be used to closely approximate the area of this circular segment? Ancient Chinese and Egyptian…

  11. Cellular immune defenses of Drosophila melanogaster.

    PubMed

    Parsons, Brendon; Foley, Edan

    2016-05-01

    Drosophila melanogaster is a widely used model for the characterization of blood cell development and function, with an array of protocols for the manipulation and visualization of fixed or live cells in vitro or in vivo. Researchers have deployed these techniques to reveal Drosophila hemocytes as a remarkably versatile cell type that engulfs apoptotic corpses; neutralizes invading parasites; seals epithelial wounds; and deposits extracellular matrix proteins. In this review, we will discuss the key features of Drosophila hemocyte development and function, and identify similarities with vertebrate counterparts. PMID:26748247

  12. Drosophila Modeling of Heritable Neurodevelopmental Disorders

    PubMed Central

    Gatto, Cheryl L.; Broadie, Kendal

    2011-01-01

    Heritable neurodevelopmental disorders are multifaceted disease conditions encompassing a wide range of symptoms including intellectual disability, cognitive dysfunction, autism and myriad other behavioral impairments. In cases where single, causative genetic defects have been identified, such as Angelman syndrome, Rett syndrome, Neurofibromatosis Type 1 and Fragile X syndrome, the classical Drosophila genetic system has provided fruitful disease models. Recent Drosophila studies have advanced our understanding of UBE3A, MECP2, NF1 and FMR1 function, respectively, in genetic, biochemical, anatomical, physiological and behavioral contexts. Investigations in Drosophila continue to provide the essential mechanistic understanding required to facilitate the conception of rational therapeutic treatments. PMID:21596554

  13. Segmental neurofibromatosis and malignancy.

    PubMed

    Dang, Julie D; Cohen, Philip R

    2010-01-01

    Segmental neurofibromatosis is an uncommon variant of neurofibromatosis type I characterized by neurofibromas and/or café-au-lait macules localized to one sector of the body. Although patients with neurofibromatosis type I have an associated increased risk of certain malignancies, malignancy has only occasionally been reported in patients with segmental neurofibromatosis. The published reports of patients with segmental neurofibromatosis who developed malignancy were reviewed and the characteristics of these patients and their cancers were summarized. Ten individuals (6 women and 4 men) with segmental neurofibromatosis and malignancy have been reported. The malignancies include malignant peripheral nerve sheath tumor (3), malignant melanoma (2), breast cancer (1), colon cancer (1), gastric cancer (1), lung cancer (1), and Hodgkin lymphoma (1). The most common malignancies in patients with segmental neurofibromatosis are derived from neural crest cells: malignant peripheral nerve sheath tumor and malignant melanoma. The incidence of malignancy in patients with segmental neurofibromatosis may approach that of patients with neurofibromatosis type I. PMID:21137621

  14. Drosophila Models of Cardiac Disease

    PubMed Central

    Piazza, Nicole; Wessells, R.J.

    2013-01-01

    The fruit fly Drosophila melanogaster has emerged as a useful model for cardiac diseases, both developmental abnormalities and adult functional impairment. Using the tools of both classical and molecular genetics, the study of the developing fly heart has been instrumental in identifying the major signaling events of cardiac field formation, cardiomyocyte specification, and the formation of the functioning heart tube. The larval stage of fly cardiac development has become an important model system for testing isolated preparations of living hearts for the effects of biological and pharmacological compounds on cardiac activity. Meanwhile, the recent development of effective techniques to study adult cardiac performance in the fly has opened new uses for the Drosophila model system. The fly system is now being used to study long-term alterations in adult performance caused by factors such as diet, exercise, and normal aging. The fly is a unique and valuable system for the study of such complex, long-term interactions, as it is the only invertebrate genetic model system with a working heart developmentally homologous to the vertebrate heart. Thus, the fly model combines the advantages of invertebrate genetics (such as large populations, facile molecular genetic techniques, and short lifespan) with physiological measurement techniques that allow meaningful comparisons with data from vertebrate model systems. As such, the fly model is well situated to make important contributions to the understanding of complicated interactions between environmental factors and genetics in the long-term regulation of cardiac performance. PMID:21377627

  15. Monoamines and sleep in Drosophila.

    PubMed

    Nall, Aleksandra; Sehgal, Amita

    2014-06-01

    Sleep is an important physiological state, but its function and regulation remain elusive. Drosophila melanogaster is a useful model organism for studying sleep because it has a well-established diurnal activity pattern, including consolidated periods of quiescence that share many characteristics with human sleep. Sleep behavior is regulated by circadian and homeostatic processes and is modulated by environmental and physiological context cues. These cues are communicated to sleep circuits by neurohormones and neuromodulators. A major class of neuromodulators, monoamines, has been found to be essential in various aspects of sleep regulation. Dopamine promotes arousal and sleep-dependent memory formation as well as daily activity. Octopamine, the insect homolog of norepinephrine, promotes wake and may play a role in circadian clock-dependent sleep and arousal. Serotonin promotes sleep and modulates circadian entrainment to light. The different monoamines each signal through multiple receptors in various brain regions in response to different conditions. How these separate circuits integrate their inputs into a single program of behavior is an open field of study for which Drosophila will continue to be a useful model. Monoamine biosynthetic pathways and receptors are conserved between flies and humans, and, thus far, their roles in modulating sleep also appear to be conserved. PMID:24886188

  16. Evaluation metrics for bone segmentation in ultrasound

    NASA Astrophysics Data System (ADS)

    Lougheed, Matthew; Fichtinger, Gabor; Ungi, Tamas

    2015-03-01

    Tracked ultrasound is a safe alternative to X-ray for imaging bones. The interpretation of bony structures is challenging as ultrasound has no specific intensity characteristic of bones. Several image segmentation algorithms have been devised to identify bony structures. We propose an open-source framework that would aid in the development and comparison of such algorithms by quantitatively measuring segmentation performance in the ultrasound images. True-positive and false-negative metrics used in the framework quantify algorithm performance based on correctly segmented bone and correctly segmented boneless regions. Ground-truth for these metrics are defined manually and along with the corresponding automatically segmented image are used for the performance analysis. Manually created ground truth tests were generated to verify the accuracy of the analysis. Further evaluation metrics for determining average performance per slide and standard deviation are considered. The metrics provide a means of evaluating accuracy of frames along the length of a volume. This would aid in assessing the accuracy of the volume itself and the approach to image acquisition (positioning and frequency of frame). The framework was implemented as an open-source module of the 3D Slicer platform. The ground truth tests verified that the framework correctly calculates the implemented metrics. The developed framework provides a convenient way to evaluate bone segmentation algorithms. The implementation fits in a widely used application for segmentation algorithm prototyping. Future algorithm development will benefit by monitoring the effects of adjustments to an algorithm in a standard evaluation framework.

  17. Conditions Affecting Social Space in Drosophila melanogaster.

    PubMed

    McNeil, Alison R; Jolley, Sam N; Akinleye, Adesanya A; Nurilov, Marat; Rouzyi, Zulekha; Milunovich, Austin J; Chambers, Moria C; Simon, Anne F

    2015-01-01

    The social space assay described here can be used to quantify social interactions of Drosophila melanogaster - or other small insects - in a straightforward manner. As we previously demonstrated (1), in a two-dimensional chamber, we first force the flies to form a tight group, subsequently allowing them to take their preferred distance from each other. After the flies have settled, we measure the distance to the closest neighbor (or social space), processing a static picture with free online software (ImageJ). The analysis of the distance to the closest neighbor allows researchers to determine the effects of genetic and environmental factors on social interaction, while controlling for potential confounding factors. Diverse factors such as climbing ability, time of day, sex, and number of flies, can modify social spacing of flies. We thus propose a series of experimental controls to mitigate these confounding effects. This assay can be used for at least two purposes. First, researchers can determine how their favorite environmental shift (such as isolation, temperature, stress or toxins) will impact social spacing (1,2). Second, researchers can dissect the genetic and neural underpinnings of this basic form of social behavior (1,3). Specifically, we used it as a diagnostic tool to study the role of orthologous genes thought to be involved in social behavior in other organisms, such as candidate genes for autism in humans (4). PMID:26575105

  18. Methodologies to determine forces on bones and muscles of body segments during exercise, employing compact sensors suitable for use in crowded space vehicles

    NASA Technical Reports Server (NTRS)

    Figueroa, Fernando

    1995-01-01

    Work under this grant was carried out by the author and by a graduate research assistant. An instrumented bicycle ergometer was implemented focusing on the stated objective: to estimate the forces exerted by each muscle of the feet, calf, and thigh of an individual while bicycling. The sensors used were light and compact. These were probes to measure muscle EMG activity, miniature accelerometers, miniature load sensors, and small encoders to measure angular positions of the pedal. A methodology was developed and implemented to completely describe the kinematics of the limbs using data from the sensors. This work has been published as a Master's Thesis by the Graduate student supported by the grant. The instrumented ergometer along with the sensors and instrumentation were tested during a KC-135 Zero-Gravity flight in July, 1994. A complete description of the system and the tests performed have been published as a report submitted to NASA Johnson Space Center. The data collected during the KC-135 flight is currently being processed so that a kinematic description of the bicycling experiment will be soon determined. A methodology to estimate the muscle forces has been formulated based on previous work. The methodology involves the use of optimization concepts so that the individual muscle forces that represent variables in dynamic equations of motion may be estimated. Optimization of a criteria (goal) function such as minimization of energy will be used along with constraint equations defined by rigid body equations of motion. Use of optimization principles is necessary, because the equations of motion alone constitute an indeterminate system of equations with respect to the large amount of muscle forces which constitute the variables in these equations. The number of variables is reduced somewhat by using forces measured by the load cells installed on the pedal. These load cells measure pressure and shear forces on the foot. The author and his collaborators at NASA

  19. Function of Lipid Storage Droplet 1 (Lsd1) in Wing Development of Drosophila melanogaster.

    PubMed

    Men, Tran Thanh; Binh, Tran Duy; Yamaguchi, Masamitsu; Huy, Nguyen Tien; Kamei, Kaeko

    2016-01-01

    Perilipins are evolutionarily conserved from Drosophila to humans, the lipid storage droplet 1 (Lsd1) is a Drosophila homolog of human perilipin 1. The function of Lsd1 as a regulator of lipolysis in Drosophila has been demonstrated, as the Lsd1 mutant causes an increase of lipid droplet size. However, the functions of this gene during development are still under investigation. In order to determine the function of Lsd1 during development, Lsd1 was knocked down in Drosophila using the GAL4-UAS system. Selective knockdown of Lsd1 in the dorsal wing disc caused an atrophied wing phenotype. The generation of reactive oxygen species in the wing pouch compartment of the Lsd1-knockdown flies was significantly higher than in the control. Immunostaining with caspase-3 antibody revealed a greater number of apoptotic cells in Lsd1-knockdown wing discs than in the control. Cell death by autophagy was also induced in the knockdown flies. Moreover, cells deprived of Lsd1 showed mitochondrial expansion and decreased ATP levels. These results strongly suggest that knockdown of Lsd1 induces mitochondrial stress and the production of reactive oxygen species that result in cell death, via apoptosis and the autophagy pathway. These results highlight the roles of Drosophila Lsd1 during wing development. PMID:27136547

  20. Function of Lipid Storage Droplet 1 (Lsd1) in Wing Development of Drosophila melanogaster

    PubMed Central

    Men, Tran Thanh; Binh, Tran Duy; Yamaguchi, Masamitsu; Huy, Nguyen Tien; Kamei, Kaeko

    2016-01-01

    Perilipins are evolutionarily conserved from Drosophila to humans, the lipid storage droplet 1 (Lsd1) is a Drosophila homolog of human perilipin 1. The function of Lsd1 as a regulator of lipolysis in Drosophila has been demonstrated, as the Lsd1 mutant causes an increase of lipid droplet size. However, the functions of this gene during development are still under investigation. In order to determine the function of Lsd1 during development, Lsd1 was knocked down in Drosophila using the GAL4-UAS system. Selective knockdown of Lsd1 in the dorsal wing disc caused an atrophied wing phenotype. The generation of reactive oxygen species in the wing pouch compartment of the Lsd1-knockdown flies was significantly higher than in the control. Immunostaining with caspase-3 antibody revealed a greater number of apoptotic cells in Lsd1-knockdown wing discs than in the control. Cell death by autophagy was also induced in the knockdown flies. Moreover, cells deprived of Lsd1 showed mitochondrial expansion and decreased ATP levels. These results strongly suggest that knockdown of Lsd1 induces mitochondrial stress and the production of reactive oxygen species that result in cell death, via apoptosis and the autophagy pathway. These results highlight the roles of Drosophila Lsd1 during wing development. PMID:27136547

  1. Hybrid image segmentation using watersheds

    NASA Astrophysics Data System (ADS)

    Haris, Kostas; Efstratiadis, Serafim N.; Maglaveras, Nicos; Pappas, Costas

    1996-02-01

    A hybrid image segmentation algorithm is proposed which combines edge- and region-based techniques through the morphological algorithm of watersheds. The algorithm consists of the following steps: (1) edge-preserving statistical noise reduction, (2) gradient approximation, (3) detection of watersheds on gradient magnitude image, and (4) hierarchical region merging (HRM) in order to get semantically meaningful segmentations. The HRM process uses the region adjacency graph (RAG) representation of the image regions. At each step, the most similar pair of regions is determined (minimum cost RAG edge), the regions are merged and the RAG is updated. Traditionally, the above is implemented by storing all the RAG edges in a priority queue (heap). We propose a significantly faster algorithm which maintains an additional graph, the most similar neighbor graph, through which the priority queue size and processing time are drastically reduced. The final segmentation is an image partition which, through the RAG, provides information that can be used by knowledge-based high level processes, i.e. recognition. In addition, this region based representation provides one-pixel wide, closed, and accurately localized contours/surfaces. Due to the small number of free parameters, the algorithm can be quite effectively used in interactive image processing. Experimental results obtained with 2D MR images are presented.

  2. Gene Regulation Networks for Modeling Drosophila Development

    NASA Technical Reports Server (NTRS)

    Mjolsness, E.

    1999-01-01

    This chapter will very briefly introduce and review some computational experiments in using trainable gene regulation network models to simulate and understand selected episodes in the development of the fruit fly, Drosophila Melanogaster.

  3. Drosophila RNAi screening in a postgenomic world

    PubMed Central

    2011-01-01

    Drosophila melanogaster has a long history as a model organism with several unique features that make it an ideal research tool for the study of the relationship between genotype and phenotype. Importantly fundamental genetic principles as well as key human disease genes have been uncovered through the use of Drosophila. The contribution of the fruit fly to science and medicine continues in the postgenomic era as cell-based Drosophila RNAi screens are a cost-effective and scalable enabling technology that can be used to quantify the contribution of different genes to diverse cellular processes. Drosophila high-throughput screens can also be used as integral part of systems-level approaches to describe the architecture and dynamics of cellular networks. PMID:21752787

  4. Drosophila Cajal bodies: accessories not included

    PubMed Central

    Matera, A. Gregory

    2006-01-01

    Cajal bodies are nuclear sites of small ribonucleoprotein (RNP) remodeling and maturation. A recent study describes the discovery of the Drosophila Cajal body, revealing some interesting insights into the subnuclear organization of RNA processing machineries among different species. PMID:16533940

  5. Targeted genome engineering techniques in Drosophila

    PubMed Central

    Beumer, Kelly J.; Carroll, Dana

    2014-01-01

    For a century, Drosophila has been a favored organism for genetic research. However, the array of materials and methods available to the Drosophila worker has expanded dramatically in the last decade. The most common gene targeting tools, zinc finger nucleases, TALENs, and RNA-guided CRISPR/Cas9, have all been adapted for use in Drosophila, both for simple mutagenesis and for gene editing via homologous recombination. For each tool, there exist a number of web sites, design applications, and delivery methods. The successful application of any of these tools also requires an understanding of methods for detecting successful genome modifications. This article provides an overview of the available gene targeting tools and their application in Drosophila. In lieu of simply providing a protocol for gene targeting, we direct the researcher to resources that will allow access to the latest research in this rapidly evolving field. PMID:24412316

  6. Circadian light-input pathways in Drosophila.

    PubMed

    Yoshii, Taishi; Hermann-Luibl, Christiane; Helfrich-Förster, Charlotte

    2016-01-01

    Light is the most important environmental cue to entrain the circadian clock in most animals. In the fruit fly Drosophila melanogaster, the light entrainment mechanisms of the clock have been well-studied. The Drosophila brain contains approximately 150 neurons that rhythmically express circadian clock genes. These neurons are called "clock neurons" and control behavioral activity rhythms. Many clock neurons express the Cryptochrome (CRY) protein, which is sensitive to UV and blue light, and thus enables clock neurons deep in the brain to directly perceive light. In addition to the CRY protein, external photoreceptors in the Drosophila eyes play an important role in circadian light-input pathways. Recent studies have provided new insights into the mechanisms that integrate these light inputs into the circadian network of the brain. In this review, we will summarize the current knowledge on the light entrainment pathways in the Drosophila circadian clock. PMID:27066180

  7. Ecdysteroid receptors in Drosophila melanogaster adult females

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ecdysteroid receptors were identified and partially characterized from total cell extracts of whole animals and dissected tissues from Drosophila melanogaster adult females. Binding studies indicated the presence of two ecdysteroid binding components having high affinity and specificity consistent w...

  8. Colour vision: parallel pathways intersect in Drosophila.

    PubMed

    Kelber, Almut; Henze, Miriam J

    2013-12-01

    In the last one hundred years, colour vision has been demonstrated in bees and many other insects. But the underlying neural wiring remained elusive. A new study on Drosophila melanogaster combining behavioural and genetic tools yields surprising insights. PMID:24309280

  9. The three-dimensional solution structure of the NK-2 homeodomain from Drosophila.

    PubMed

    Tsao, D H; Gruschus, J M; Wang, L H; Nirenberg, M; Ferretti, J A

    1995-08-11

    We describe the NMR determination of the three-dimensional structure of a 77 amino acid residue protein, which consists of the 60 residue NK-2 homeodomain from Drosophila melanogaster and adjacent amino acid residues. The NK-2 homeodomain protein is part of a 723 amino acid residue protein which is expressed early in embryonic development in part of the central nervous system. NK-2 was characterized using both a natural abundance and a uniformly 15N enriched sample by two-dimensional and three-dimensional NMR experiments. The average root-mean-square deviation for 30 structures for residues 8 to 53 is 0.40 A for the backbone heavy-atoms and 0.72 A for the backbone and side-chain heavy-atoms. These structures were obtained from 986 NOE-derived upper and lower bound restraints. The three-dimensional structure contains three helices which consist of homeodomain amino acid residues 10 to 22, 28 to 38 and 42 to 52, as well as a turn between helix II and III, characteristic of homeodomains. Residues 53 to 60 of the DNA recognition helix are not fully ordered in the absence of DNA. In the free state this segment adopts a flexible but helix-like structure between residues 53 and 56 and is disordered from residues 57 to 60 although, as shown previously, the helix elongates by eight residues upon binding to DNA. The role of variable residues 52, 54 and 56 in determining the structure and flexibility of the recognition helix, as well as the stability of the NK-2 homeodomain as manifested by its thermal denaturation, are discussed. PMID:7643404

  10. Genome-wide mapping of in vivo targets of the Drosophila transcription factor Kruppel.

    PubMed

    Matyash, Alexey; Chung, Ho-Ryun; Jäckle, Herbert

    2004-07-16

    Krüppel (Kr), a member of the gap class of Drosophila segmentation genes, encodes a DNA binding zinc finger-type transcription factor. In addition to its segmentation function at the blastoderm stage, Krüppel also plays a critical role in organ formation during later stages of embryogenesis. To systematically identify in vivo target genes of Krüppel, we isolated DNA fragments from the Krüppel-associated portion of chromatin and used them to find and map Krüppel-dependent cis-acting regulatory sites in the Drosophila genome. We show that Krüppel binding sites are not enriched in Krüppel-associated chromatin and that the clustering of Krüppel binding sites, as found in the cis-acting elements of Krüppel-dependent segmentation genes used for in silico searches of Krüppel target genes, is not a prerequisite for the in vivo binding of Krüppel to its regulatory elements. Results obtained with the newly identified target gene ken and barbie (ken) indicate that Krüppel represses transcription and thereby restricts the spatial expression pattern of ken during blastoderm and gastrulation. PMID:15131112

  11. Identification of excitatory premotor interneurons which regulate local muscle contraction during Drosophila larval locomotion.

    PubMed

    Hasegawa, Eri; Truman, James W; Nose, Akinao

    2016-01-01

    We use Drosophila larval locomotion as a model to elucidate the working principles of motor circuits. Larval locomotion is generated by rhythmic and sequential contractions of body-wall muscles from the posterior to anterior segments, which in turn are regulated by motor neurons present in the corresponding neuromeres. Motor neurons are known to receive both excitatory and inhibitory inputs, combined action of which likely regulates patterned motor activity during locomotion. Although recent studies identified candidate inhibitory premotor interneurons, the identity of premotor interneurons that provide excitatory drive to motor neurons during locomotion remains unknown. In this study, we searched for and identified two putative excitatory premotor interneurons in this system, termed CLI1 and CLI2 (cholinergic lateral interneuron 1 and 2). These neurons were segmentally arrayed and activated sequentially from the posterior to anterior segments during peristalsis. Consistent with their being excitatory premotor interneurons, the CLIs formed GRASP- and ChAT-positive putative synapses with motoneurons and were active just prior to motoneuronal firing in each segment. Moreover, local activation of CLI1s induced contraction of muscles in the corresponding body segments. Taken together, our results suggest that the CLIs directly activate motoneurons sequentially along the segments during larval locomotion. PMID:27470675

  12. Identification of excitatory premotor interneurons which regulate local muscle contraction during Drosophila larval locomotion

    PubMed Central

    Hasegawa, Eri; Truman, James W.; Nose, Akinao

    2016-01-01

    We use Drosophila larval locomotion as a model to elucidate the working principles of motor circuits. Larval locomotion is generated by rhythmic and sequential contractions of body-wall muscles from the posterior to anterior segments, which in turn are regulated by motor neurons present in the corresponding neuromeres. Motor neurons are known to receive both excitatory and inhibitory inputs, combined action of which likely regulates patterned motor activity during locomotion. Although recent studies identified candidate inhibitory premotor interneurons, the identity of premotor interneurons that provide excitatory drive to motor neurons during locomotion remains unknown. In this study, we searched for and identified two putative excitatory premotor interneurons in this system, termed CLI1 and CLI2 (cholinergic lateral interneuron 1 and 2). These neurons were segmentally arrayed and activated sequentially from the posterior to anterior segments during peristalsis. Consistent with their being excitatory premotor interneurons, the CLIs formed GRASP- and ChAT-positive putative synapses with motoneurons and were active just prior to motoneuronal firing in each segment. Moreover, local activation of CLI1s induced contraction of muscles in the corresponding body segments. Taken together, our results suggest that the CLIs directly activate motoneurons sequentially along the segments during larval locomotion. PMID:27470675

  13. A Drosophila muscle-specific gene related to the mouse quaking locus.

    PubMed

    Fyrberg, C; Becker, J; Barthmaier, P; Mahaffey, J; Fyrberg, E

    1997-09-15

    We have characterized a novel muscle-specific gene of Drosophila melanogaster, defined by enhancer trap strain 24B of Brand and Perrimon (1993). We show that transcripts of the gene accumulate within presumptive mesoderm and persist within developing muscles, strongly suggesting that the encoded protein is involved in muscle cell determination and differentiation. cDNA sequences reveal that the Drosophila protein is similar to quaking (64% identity over 210 amino acids), a protein essential for mouse embryogenesis, and gld-1 (53% identity over 162 amino acids) a germ-line-specific tumor suppressing protein of the nematode, Caenorhabditis elegans. We demonstrate that the Drosophila gene resides within the 93F chromosome subdivision, and describe its physical map. Finally, we have used the gene, which we have named quaking-related 93F (qkr93F), to identify a family of closely related KH domains. PMID:9332381

  14. Low level image segmentation: an expert system.

    PubMed

    Nazif, A M; Levine, M D

    1984-05-01

    A major problem in robotic vision is the segmentation of images of natural scenes in order to understand their content. This paper presents a new solution to the image segmentation problem that is based on the design of a rule-based expert system. General knowledge about low level properties of processes employ the rules to segment the image into uniform regions and connected lines. In addition to the knowledge rules, a set of control rules are also employed. These include metarules that embody inferences about the order in which the knowledge rules are matched. They also incorporate focus of attention rules that determine the path of processing within the image. Furthermore, an additional set of higher level rules dynamically alters the processing strategy. This paper discusses the structure and content of the knowledge and control rules for image segmentation. PMID:21869225

  15. Bootstrapping structured page segmentation

    NASA Astrophysics Data System (ADS)

    Ma, Huanfeng; Doermann, David S.

    2003-01-01

    In this paper, we present an approach to the bootstrap learning of a page segmentation model. The idea evolves from attempts to segment dictionaries that often have a consistent page structure, and is extended to the segmentation of more general structured documents. In cases of highly regular structure, the layout can be learned from examples of only a few pages. The system is first trained using a small number of samples, and a larger test set is processed based on the training result. After making corrections to a selected subset of the test set, these corrected samples are combined with the original training samples to generate bootstrap samples. The newly created samples are used to retrain the system, refine the learned features and resegment the test samples. This procedure is applied iteratively until the learned parameters are stable. Using this approach, we do not need to initially provide a large set of training samples. We have applied this segmentation to many structured documents such as dictionaries, phone books, spoken language transcripts, and obtained satisfying segmentation performance.

  16. Scorpion image segmentation system

    NASA Astrophysics Data System (ADS)

    Joseph, E.; Aibinu, A. M.; Sadiq, B. A.; Bello Salau, H.; Salami, M. J. E.

    2013-12-01

    Death as a result of scorpion sting has been a major public health problem in developing countries. Despite the high rate of death as a result of scorpion sting, little report exists in literature of intelligent device and system for automatic detection of scorpion. This paper proposed a digital image processing approach based on the floresencing characteristics of Scorpion under Ultra-violet (UV) light for automatic detection and identification of scorpion. The acquired UV-based images undergo pre-processing to equalize uneven illumination and colour space channel separation. The extracted channels are then segmented into two non-overlapping classes. It has been observed that simple thresholding of the green channel of the acquired RGB UV-based image is sufficient for segmenting Scorpion from other background components in the acquired image. Two approaches to image segmentation have also been proposed in this work, namely, the simple average segmentation technique and K-means image segmentation. The proposed algorithm has been tested on over 40 UV scorpion images obtained from different part of the world and results obtained show an average accuracy of 97.7% in correctly classifying the pixel into two non-overlapping clusters. The proposed 1system will eliminate the problem associated with some of the existing manual approaches presently in use for scorpion detection.

  17. Cooperative processes in image segmentation

    NASA Technical Reports Server (NTRS)

    Davis, L. S.

    1982-01-01

    Research into the role of cooperative, or relaxation, processes in image segmentation is surveyed. Cooperative processes can be employed at several levels of the segmentation process as a preprocessing enhancement step, during supervised or unsupervised pixel classification and, finally, for the interpretation of image segments based on segment properties and relations.

  18. Two retrotransposons maintain telomeres in Drosophila

    PubMed Central

    Pardue, M.-L.; Rashkova, S.; Casacuberta, E.; DeBaryshe, P.G.; George, J. A.; Traverse, K.L.

    2005-01-01

    Telomeres across the genus Drosophila are maintained, not by telomerase, but by two non-LTR retrotransposons, HeT-A and TART, that transpose specifically to chromosome ends. Successive transpositions result in long head-to-tail arrays of these elements. Thus Drosophila telomeres, like those produced by telomerase, consist of repeated sequences reverse transcribed from RNA templates. The Drosophila repeats, complete and 5′-truncated copies of HeT-A and TART, are more complex than telomerase repeats; nevertheless these evolutionary variants have functional similarities to the more common telomeres. Like other telomeres, the Drosophila arrays are dynamic, fluctuating around an average length that can be changed by changes in the genetic background. Several proteins that interact with telomeres in other species have been found to have homologues that interact with Drosophila telomeres. Although they have hallmarks of non-LTR retrotransposons, HeT-A and TART appear to have a special relationship to Drosophila. Their Gag proteins are efficiently transported into diploid nuclei where HeT-A Gag recruits TART Gag to chromosome ends. Gags of other non-LTR elements remain predominantly in the cytoplasm. These studies provide intriguing evolutionary links between telomeres and retrotransposable elements. PMID:16132810

  19. A Drosophila Model for Screening Antiobesity Agents

    PubMed Central

    Men, Tran Thanh; Thanh, Duong Ngoc Van; Yamaguchi, Masamitsu; Suzuki, Takayoshi; Hattori, Gen; Arii, Masayuki; Huy, Nguyen Tien; Kamei, Kaeko

    2016-01-01

    Although triacylglycerol, the major component for lipid storage, is essential for normal physiology, its excessive accumulation causes obesity in adipose tissue and is associated with organ dysfunction in nonadipose tissue. Here, we focused on the Drosophila model to develop therapeutics for preventing obesity. The brummer (bmm) gene in Drosophila melanogaster is known to be homologous with human adipocyte triglyceride lipase, which is related to the regulation of lipid storage. We established a Drosophila model for monitoring bmm expression by introducing the green fluorescent protein (GFP) gene as a downstream reporter of the bmm promoter. The third-instar larvae of Drosophila showed the GFP signal in all tissues observed and specifically in the salivary gland nucleus. To confirm the relationship between bmm expression and obesity, the effect of oral administration of glucose diets on bmm promoter activity was analyzed. The Drosophila flies given high-glucose diets showed higher lipid contents, indicating the obesity phenotype; this was suggested by a weaker intensity of the GFP signal as well as reduced bmm mRNA expression. These results demonstrated that the transgenic Drosophila model established in this study is useful for screening antiobesity agents. We also report the effects of oral administration of histone deacetylase inhibitors and some vegetables on the bmm promoter activity. PMID:27247940

  20. Homoeosis in Drosophila: a description of the Polycomb lethal syndrome.

    PubMed

    Denell, R E; Frederick, R D

    1983-05-01

    Adults heterozygous for dominant mutations at the haploabnormal Polycomb (Pc) locus display many homoeotic transformations. E. B. Lewis (1978) first described the cuticular morphology of lethal embryos homozygous for Polycomb mutant alleles, and suggested that the Pc+ gene product acts as a repressor of genes in the Bithorax gene complex. In the present work, we have further examined the Polycomb lethal syndrome by phase-contrast and scanning electron microscopy of whole mounts, and show that the phenotype is more complex than hitherto realized. Many cuticular features characteristic of more anterior body segments are partially or completely transformed to resemble those of more caudal segments; the posteriormost homoeotic features which develop are those of the 8th abdominal segment (rather than the 9th or 10th). Involution and dorsal closure of the head are usually incomplete, and the labium and dorsal head regularly develop cuticular features normally characteristic of abdominal segments. Not all homoeotic alterations are caudal, however, and embryos also display partial transformations of the 9th (or 10th) abdominal segment to 8th and of meso- and metathorax to prothorax. It should be emphasized that Polycomb mutations do not homoeotically transform segments as a whole; various cuticular markers differ considerably in their relative probability and extent of transformation, and a single segment may display features normally specific to several different segments. We suggest that these mutations result in an instability in the transmission of determined states. PMID:6404678

  1. Temporal pattern of the posterior expression of Wingless in Drosophila blastoderm

    PubMed Central

    Vorwald-Denholtz, Peggy P.; De Robertis, Edward M.

    2011-01-01

    In most animals, the Antero-Posterior (A-P) axis requires a gradient of Wnt signaling. Wnts are expressed posteriorly in many vertebrate and invertebrate embryos, forming a gradient of canonical Wnt/β-Catenin activity that is highest in the posterior and lowest in the anterior. One notable exception to this evolutionary conservation is in the Drosophila embryo, in which the A-P axis is established by early transcription factors of maternal origin. Despite this initial axial establishment, Drosophila still expresses Wingless (Wg), the main Drosophila Wnt homologue, in a strong posterior band early in embryogenesis. Since its discovery 30 years ago this posterior band of Wg has been largely ignored. In this study, we re-examined the onset of expression of the Wg posterior band in relation to the expression of Wg in other segments, and compared the timing of its expression to that of axial regulators such as gap and pair-rule genes. It was found that the posterior band of Wg is first detected in blastoderm at mid nuclear cycle 14, before the segment-polarity stripes of Wg are formed in other segments. The onset of the posterior band of Wg expression was preceded by that of the gap gene products Hunchback (hb) and Krüppel (Kr), and the pair-rule protein Even-skipped (Eve). Although the function of the posterior band of Wg was not analyzed in this study, we note that in temperature-sensitive Wg mutants, in which Wg is not properly secreted, the posterior band of Wg expression is diminished in strength, indicating a positive feedback loop required for Wg robust expression at the cellular blastoderm stage. We propose that this early posterior expression could play a role in the refinement of A-P patterning. PMID:21821151

  2. Postharvest treatment of strawberries with methyl bromide to control spotted wing drosophila, Drosophila suzukii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spotted wing drosophila (SWD), Drosophila suzukii, is a pest of concern to countries that import strawberries from California USA. The purpose of this investigation was to verify elimination of SWD in California-grown fresh strawberries under conditions consistent with export to Australia; a 3-h fum...

  3. Current Recommendations for Managing Spotted Wing Drosophila (SWD), Drosophila suzukii, in PNW Strawberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The spotted wing Drosophila (SWD), Drosophila suzukii, was reported in the Pacific Northwest (Oregon, Washington, British Columbia) in 2009. The fly is able to oviposit directly into intact ripe and ripening fruit, so it is of great economic concern to the small fruit industries in region. Fruit i...

  4. Current Recommendations for Managing Spotted Wing Drosophila (SWD), Drosophila suzukii, in PNW Blueberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The spotted wing Drosophila (SWD), Drosophila suzukii, was reported in the Pacific Northwest (Oregon, Washington, British Columbia) in 2009. The fly is able to oviposit directly into intact ripe and ripening fruit, so it is of great economic concern to the small fruit industries in region. Fruit i...

  5. Current Recommendations for Managing Spotted Wing Drosophila (SWD), Drosophila suzukii, in PNW Caneberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The spotted wing Drosophila (SWD), Drosophila suzukii, was reported in the Pacific Northwest (Oregon, Washington, British Columbia) in 2009. The fly is able to oviposit directly into intact ripe and ripening fruit, so it is of great economic concern to the small fruit industries in region. Fruit i...

  6. The susceptibility of small fruits and cherries to Spotted Wing Drosophila, Drosophila suzukii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: The Spotted Wing Drosophila (SWD), Drosophila suzukii Matsumura, is native to Asia and has been detected in the North American mainland and Europe in 2008-10. SWD is a serious economic pest because it lays eggs within ripening fruit before harvest which can lead to crop loss. The aim ...

  7. Behavioral and antennal responses of spotted wing drosophila, drosophila suzukii, to volatiles from fruit extracts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Native to Southeast Asia, the spotted wing drosophila, Drosophila suzukii, has become a serious pest of soft-skinned fruit crops since its introduction into North America and Europe in 2008. Current monitoring strategies use baits based on fermentation products; however, to date, no fruit-based vola...

  8. Invasion biology of Spotted Wing Drosophila (Drosophila suzukii): a global perspective and future priorities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Asian vinegar fly species Drosophila suzukii (spotted-wing Drosophila or SWD) has emerged as an important invasive insect pest of small and stone fruits in both the Americas and Europe since the late 2000’s. While research efforts have rapidly progressed in Asia, North America, and Europe over ...

  9. Spotted wing drosophila, Drosophila suzukii (Matsumura)(Diptera: drosophilidae), trapped with combinations of wines and vinegars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field trapping experiments evaluated wine and vinegar baits for spotted wing drosophila flies, Drosophila suzukii (Matsumura), and assessed variance in biat attractiveness with wit type, vinegar type, and bait age. A mixture of apple cider vinegar and a Merlot wine attracted more flies than a mixtur...

  10. Structural implications of weak Ca2+ block in Drosophila cyclic nucleotide-gated channels.

    PubMed

    Lam, Yee Ling; Zeng, Weizhong; Derebe, Mehabaw Getahun; Jiang, Youxing

    2015-09-01

    Calcium permeability and the concomitant calcium block of monovalent ion current ("Ca(2+) block") are properties of cyclic nucleotide-gated (CNG) channel fundamental to visual and olfactory signal transduction. Although most CNG channels bear a conserved glutamate residue crucial for Ca(2+) block, the degree of block displayed by different CNG channels varies greatly. For instance, the Drosophila melanogaster CNG channel shows only weak Ca(2+) block despite the presence of this glutamate. We previously constructed a series of chimeric channels in which we replaced the selectivity filter of the bacterial nonselective cation channel NaK with a set of CNG channel filter sequences and determined that the resulting NaK2CNG chimeras displayed the ion selectivity and Ca(2+) block properties of the parent CNG channels. Here, we used the same strategy to determine the structural basis of the weak Ca(2+) block observed in the Drosophila CNG channel. The selectivity filter of the Drosophila CNG channel is similar to that of most other CNG channels except that it has a threonine at residue 318 instead of a proline. We constructed a NaK chimera, which we called NaK2CNG-Dm, which contained the Drosophila selectivity filter sequence. The high resolution structure of NaK2CNG-Dm revealed a filter structure different from those of NaK and all other previously investigated NaK2CNG chimeric channels. Consistent with this structural difference, functional studies of the NaK2CNG-Dm chimeric channel demonstrated a loss of Ca(2+) block compared with other NaK2CNG chimeras. Moreover, mutating the corresponding threonine (T318) to proline in Drosophila CNG channels increased Ca(2+) block by 16 times. These results imply that a simple replacement of a threonine for a proline in Drosophila CNG channels has likely given rise to a distinct selectivity filter conformation that results in weak Ca(2+) block. PMID:26283200

  11. Phasing a segmented telescope

    NASA Astrophysics Data System (ADS)

    Paykin, Irina; Yacobi, Lee; Adler, Joan; Ribak, Erez N.

    2015-02-01

    A crucial part of segmented or multiple-aperture systems is control of the optical path difference between the segments or subapertures. In order to achieve optimal performance we have to phase subapertures to within a fraction of the wavelength, and this requires high accuracy of positioning for each subaperture. We present simulations and hardware realization of a simulated annealing algorithm in an active optical system with sparse segments. In order to align the optical system we applied the optimization algorithm to the image itself. The main advantage of this method over traditional correction methods is that wave-front-sensing hardware and software are no longer required, making the optical and mechanical system much simpler. The results of simulations and laboratory experiments demonstrate the ability of this optimization algorithm to correct both piston and tip-tilt errors.

  12. Segmented annular combustor

    DOEpatents

    Reider, Samuel B.

    1979-01-01

    An industrial gas turbine engine includes an inclined annular combustor made up of a plurality of support segments each including inner and outer walls of trapezoidally configured planar configuration extents and including side flanges thereon interconnected by means of air cooled connector bolt assemblies to form a continuous annular combustion chamber therebetween and wherein an air fuel mixing chamber is formed at one end of the support segments including means for directing and mixing fuel within a plenum and a perforated header plate for directing streams of air and fuel mixture into the combustion chamber; each of the outer and inner walls of each of the support segments having a ribbed lattice with tracks slidably supporting porous laminated replaceable panels and including pores therein for distributing combustion air into the combustion chamber while cooling the inner surface of each of the panels by transpiration cooling thereof.

  13. Phasing a segmented telescope.

    PubMed

    Paykin, Irina; Yacobi, Lee; Adler, Joan; Ribak, Erez N

    2015-02-01

    A crucial part of segmented or multiple-aperture systems is control of the optical path difference between the segments or subapertures. In order to achieve optimal performance we have to phase subapertures to within a fraction of the wavelength, and this requires high accuracy of positioning for each subaperture. We present simulations and hardware realization of a simulated annealing algorithm in an active optical system with sparse segments. In order to align the optical system we applied the optimization algorithm to the image itself. The main advantage of this method over traditional correction methods is that wave-front-sensing hardware and software are no longer required, making the optical and mechanical system much simpler. The results of simulations and laboratory experiments demonstrate the ability of this optimization algorithm to correct both piston and tip-tilt errors. PMID:25768631

  14. Head segmentation in vertebrates

    PubMed Central

    Kuratani, Shigeru; Schilling, Thomas

    2008-01-01

    Classic theories of vertebrate head segmentation clearly exemplify the idealistic nature of comparative embryology prior to the 20th century. Comparative embryology aimed at recognizing the basic, primary structure that is shared by all vertebrates, either as an archetype or an ancestral developmental pattern. Modern evolutionary developmental (Evo-Devo) studies are also based on comparison, and therefore have a tendency to reduce complex embryonic anatomy into overly simplified patterns. Here again, a basic segmental plan for the head has been sought among chordates. We convened a symposium that brought together leading researchers dealing with this problem, in a number of different evolutionary and developmental contexts. Here we give an overview of the outcome and the status of the field in this modern era of Evo-Devo. We emphasize the fact that the head segmentation problem is not fully resolved, and we discuss new directions in the search for hints for a way out of this maze. PMID:20607135

  15. Transcriptomic profile of Drosophila melanogaster larval midgut and responses to oxidative stress

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oligoarray analysis was used to determine the number and nature of genes expressed in third-instar Drosophila melanogaster larval midguts. The majority of transcripts were associated with protein synthesis and metabolism. Serine proteases were the main proteolytic enzymes detected. Some 40% of th...

  16. Humidity implications for populations of Drosophila suzukii (Diptera: Drosophilidae) on blueberry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Temperature and humidity affect insect physiology, survival, fecundity, reproductive status and behavior. Drosophila suzukii is an invasive pest of soft-skinned fruit and can cause severe economic losses in a wide range of susceptible crops. This study was conducted on blueberries to determine the e...

  17. The Impact of Odor--Reward Memory on Chemotaxis in Larval "Drosophila"

    ERIC Educational Resources Information Center

    Schleyer, Michael; Reid, Samuel F.; Pamir, Evren; Saumweber, Timo; Paisios, Emmanouil; Davies, Alexander; Gerber, Bertram; Louis, Matthieu

    2015-01-01

    How do animals adaptively integrate innate with learned behavioral tendencies? We tackle this question using chemotaxis as a paradigm. Chemotaxis in the "Drosophila" larva largely results from a sequence of runs and oriented turns. Thus, the larvae minimally need to determine (i) how fast to run, (ii) when to initiate a turn, and (iii)…

  18. Evaluating Drosophila suzukii (Diptera: Drosophilidae) immunomarking for mark-capture research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Drosophila suzukii Matsumura readily utilizes wild Himalayan blackberry, Rubus armeniacus Focke as a host and is suspected of invading berry and stone fruit crops from field margins containing this invasive weed. This study was conducted to determine: (1) protein mark (10% chicken egg whites [albumi...

  19. Tetrahydropterin as a possible natural cofactor in the drosophila phenylalanine hydroxylation system

    SciTech Connect

    Bel, Y.; Jacobson, K.B.; Ferre, J. . Dept. of Genetics; Oak Ridge National Lab., TN; Valencia Univ. . Dept. of Genetics)

    1989-01-01

    The aim of the present work is the study of phenylalanine hydroxylase (PH) activity of Drosophila melanogaster wild type with different cofactors: the two natural occurring tetrahydropteridines (BH{sub 4} and PH{sub 4}) and the synthetic 6,7-dimethyltetrahydropterin (DMPH{sub 4}), as well as the determination of this activity at different developmental stages. 7 refs., 2 figs.

  20. The role of reduced oxygen in the developmental physiology of growth and metamorphosis initiation in Drosophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rearing oxygen level is known to affect final body size in a variety of insects, but the physiological mechanisms by which oxygen affects size are incompletely understood. In Manduca and Drosophila, the larval size at which metamorphosis is initiated largely determines adult size, and metamorphosis ...

  1. Systematic expression and loss-of-function analysis defines spatially restricted requirements for Drosophila RhoGEFs and RhoGAPs in leg morphogenesis

    PubMed Central

    Greenberg, Lina; Hatini, Victor

    2010-01-01

    The Drosophila leg imaginal disc consists of a peripheral region that contributes to adult body wall, and a central region that forms the leg proper. While the patterning signals and transcription factors that determine the identity of adult structures have been identified, the mechanisms that determine the shape of these structures remain largely unknown. The family of Rho GTPases, which consists of 7 members in flies, modulates cell adhesion, actomyosin contractility, protrusive membrane activity, and cell-matrix adhesion to generate mechanical forces that shape adult structures. The Rho GTPases are ubiquitously expressed and it remains unclear how they orchestrate morphogenetic events. The Rho guanine nucleotide exchange factors (RhoGEFs) and Rho GTPase activating proteins (RhoGAPs), which respectively activate and deactivate corresponding Rho GTPases, have been proposed to regulate the activity of Rho signaling cascades in specific spatiotemporal patterns to orchestrate morphogenetic events. Here we identify restricted expression of 12 of the 20 RhoGEFs and 10 of the 22 Rho RhoGAPs encoded in Drosophila during metamorphosis. Expression of a subset of each family of RhoGTPase regulators was restricted to motile cell populations including tendon, muscle, trachea, and peripodial stalk cells. A second subset was restricted either to all presumptive joints or only to presumptive tarsal joints. Depletion of individual RhoGEFs and RhoGAPs in the epithelium of the disc proper identified several joint-specific genes, which act downstream of segmental patterning signals to control epithelial morphogenesis. Our studies provide a framework with which to understand how Rho signaling cascades orchestrate complex morphogenetic events in multicellular organisms, and evidence that patterning signals regulate these cascades to control apical constriction and epithelial invagination at presumptive joints. PMID:20851182

  2. Investigating Spermatogenesis in Drosophila melanogaster

    PubMed Central

    Demarco, Rafael S.; Eikenes, Åsmund H.; Haglund, Kaisa; Jones, D. Leanne

    2014-01-01

    The process of spermatogenesis in Drosophila melanogaster provides a powerful model system to probe a variety of developmental and cell biological questions, such as the characterization of mechanisms that regulate stem cell behavior, cytokinesis, meiosis, and mitochondrial dynamics. Classical genetic approaches, together with binary expression systems, FRT-mediated recombination, and novel imaging systems to capture single cell behavior, are rapidly expanding our knowledge of the molecular mechanisms regulating all aspects of spermatogenesis. This methods chapter provides a detailed description of the system, a review of key questions chapter that have been addressed or remain unanswered thus far, and an introduction to tools and techniques available to probe each stage of spermatogenesis. PMID:24798812

  3. MicroRNA targets in Drosophila

    PubMed Central

    Enright, Anton J; John, Bino; Gaul, Ulrike; Tuschl, Thomas; Sander, Chris; Marks, Debora S

    2004-01-01

    Background The recent discoveries of microRNA (miRNA) genes and characterization of the first few target genes regulated by miRNAs in Caenorhabditis elegans and Drosophila melanogaster have set the stage for elucidation of a novel network of regulatory control. We present a computational method for whole-genome prediction of miRNA target genes. The method is validated using known examples. For each miRNA, target genes are selected on the basis of three properties: sequence complementarity using a position-weighted local alignment algorithm, free energies of RNA-RNA duplexes, and conservation of target sites in related genomes. Application to the D. melanogaster, Drosophila pseudoobscura and Anopheles gambiae genomes identifies several hundred target genes potentially regulated by one or more known miRNAs. Results These potential targets are rich in genes that are expressed at specific developmental stages and that are involved in cell fate specification, morphogenesis and the coordination of developmental processes, as well as genes that are active in the mature nervous system. High-ranking target genes are enriched in transcription factors two-fold and include genes already known to be under translational regulation. Our results reaffirm the thesis that miRNAs have an important role in establishing the complex spatial and temporal patterns of gene activity necessary for the orderly progression of development and suggest additional roles in the function of the mature organism. In addition the results point the way to directed experiments to determine miRNA functions. Conclusions The emerging combinatorics of miRNA target sites in the 3' untranslated regions of messenger RNAs are reminiscent of transcriptional regulation in promoter regions of DNA, with both one-to-many and many-to-one relationships between regulator and target. Typically, more than one miRNA regulates one message, indicative of cooperative translational control. Conversely, one miRNA may have

  4. Drosophila Melanogaster Show a Threshold Effect in Response to Radiation

    PubMed Central

    Antosh, Michael; Fox, David; Hasselbacher, Thomas; Lanou, Robert; Neretti, Nicola; Cooper, Leon N.

    2014-01-01

    We investigate the biological effects of radiation using adult Drosophila melanogaster as a model organism, focusing on gene expression and lifespan analysis to determine the effect of different radiation doses. Our results support a threshold effect in response to radiation: no effect on lifespan and no permanent effect on gene expression is seen at incident radiation levels below 100 J/kg. We also find that it is more appropriate to compare radiation effects in flies using the absorbed energy rather than incident radiation levels. PMID:25552957

  5. Semiautomatic and rapid quantification of heartbeat parameters in Drosophila using optical coherence tomography imaging

    NASA Astrophysics Data System (ADS)

    Guo, Shou-Yuan; Liao, Fang-Tsu; Su, Ming-Tsan; Chang, Cheng-Yi; Su, Hong-Ren; Huang, Jyun-Cin; Kuo, Wen-Chuan

    2013-02-01

    We report a semiautomatic algorithm that is specialized for rapid analysis of beat-to-beat contraction-relaxation parameters of the heart in Drosophila. The presented algorithm adapts the general graph theoretical image segmentation algorithm and a histogram-based thresholding algorithm, which can measure many cardiac parameters, including heart rate, heart period, diastolic and systolic intervals, and end-diastolic and end-systolic areas. Additionally, dynamic cardiac functions, such as arrhythmia index and percent fractional shortening, can be automatically calculated for all the recorded heartbeats over significant periods of time.

  6. The use of a mutationally unstable X-chromosome in Drosophila melanogaster for mutagenicity testing.

    PubMed

    Rasmuson, B; Svahlin, H; Rasmuson, A; Montell, I; Olofsson, H

    1978-08-01

    Somatic eye-colour mutations in an unstable genetic system, caused by a transposable element in the white locus of the X-chromosome in Drosophila melanogaster, is suggested as an assay system for mutagenicity testing. The system is evaluated by comparison with a corresponding system in a stable X-chromosome. Its sensitivity is confirmed with X-ray and EMS treatment, and it is found to be confined to the specific segment of the X-chromosome where the transposable element is localized. PMID:97525

  7. A probabilistic level set formulation for interactive organ segmentation

    NASA Astrophysics Data System (ADS)

    Cremers, Daniel; Fluck, Oliver; Rousson, Mikael; Aharon, Shmuel

    2007-03-01

    Level set methods have become increasingly popular as a framework for image segmentation. Yet when used as a generic segmentation tool, they suffer from an important drawback: Current formulations do not allow much user interaction. Upon initialization, boundaries propagate to the final segmentation without the user being able to guide or correct the segmentation. In the present work, we address this limitation by proposing a probabilistic framework for image segmentation which integrates input intensity information and user interaction on equal footings. The resulting algorithm determines the most likely segmentation given the input image and the user input. In order to allow a user interaction in real-time during the segmentation, the algorithm is implemented on a graphics card and in a narrow band formulation.

  8. Study of dosage compensation in Drosophila.

    PubMed Central

    Chiang, Pei-Wen; Kurnit, David M

    2003-01-01

    Using a sensitive RT-QPCR assay, we analyzed the regulatory effects of sex and different dosage compensation mutations in Drosophila. To validate the assay, we showed that regulation for several genes indeed varied with the number of functional copies of that gene. We then confirmed that dosage compensation occurred for most genes we examined in male and female flies. Finally, we examined the effects on regulation of several genes in the MSL pathway, presumed to be involved in sex-dependent determination of regulation. Rather than seeing global alterations of either X chromosomal or autosomal genes, regulation of genes on either the X chromosome or the autosomes could be elevated, depressed, or unaltered between sexes in unpredictable ways for the various MSL mutations. Relative dosage for a given gene between the sexes could vary at different developmental times. Autosomal genes often showed deranged regulatory levels, indicating they were in pathways perturbed by X chromosomal changes. As exemplified by the BR-C locus and its dependent Sgs genes, multiple genes in a given pathway could exhibit coordinate regulatory modulation. The variegated pattern shown for expression of both X chromosomal and autosomal loci underscores the complexity of gene expression so that the phenotype of MSL mutations does not reflect only simple perturbations of genes on the X chromosome. PMID:14668373

  9. Patterns of Hermes transposition in Drosophila melanogaster.

    PubMed

    Guimond, N; Bideshi, D K; Pinkerton, A C; Atkinson, P W; O'Brochta, D A

    2003-03-01

    Transposable elements are being developed as tools for genomics and for the manipulation of insect genotypes for the purposes of biological control. An understanding of their transposition behavior will facilitate the use of these elements. The behavior of an autonomous Hermes transposable element from Musca domestica in the soma and germ-line of Drosophila melanogaster was investigated using the method of transposon display. In the germ-line, Hermes transposed at a rate of approximately 0.03 jumps per element per generation. Within the soma Hermes exhibited markedly non-random patterns of integration. Certain regions of the genome were distinctly preferred over others as integration targets, while other regions were underrepresented among the integration sites used. One particular site accounted for 4.4% of the transpositions recovered in this experiment, all of which were located within a 2.5-kb region of the actin5C promoter. This region was also present within the Hermes element itself, suggesting that this clustering is an example of transposable element "homing". Clusters of integration sites were also observed near the original donor sites; these represent examples of local hopping. The information content (sequence specificity) of the 8-bp target site was low, and the consensus target site resembles that determined from plasmid-based integration assays. PMID:12655404

  10. Parallel geographic variation in Drosophila melanogaster.

    PubMed

    Reinhardt, Josie A; Kolaczkowski, Bryan; Jones, Corbin D; Begun, David J; Kern, Andrew D

    2014-05-01

    Drosophila melanogaster, an ancestrally African species, has recently spread throughout the world, associated with human activity. The species has served as the focus of many studies investigating local adaptation relating to latitudinal variation in non-African populations, especially those from the United States and Australia. These studies have documented the existence of shared, genetically determined phenotypic clines for several life history and morphological traits. However, there are no studies designed to formally address the degree of shared latitudinal differentiation at the genomic level. Here we present our comparative analysis of such differentiation. Not surprisingly, we find evidence of substantial, shared selection responses on the two continents, probably resulting from selection on standing ancestral variation. The polymorphic inversion In(3R)P has an important effect on this pattern, but considerable parallelism is also observed across the genome in regions not associated with inversion polymorphism. Interestingly, parallel latitudinal differentiation is observed even for variants that are not particularly strongly differentiated, which suggests that very large numbers of polymorphisms are targets of spatially varying selection in this species. PMID:24610860

  11. Drosophila as a model to study the role of blood cells in inflammation, innate immunity and cancer

    PubMed Central

    Wang, Lihui; Kounatidis, Ilias; Ligoxygakis, Petros

    2014-01-01

    Drosophila has a primitive yet effective blood system with three types of haemocytes which function throughout different developmental stages and environmental stimuli. Haemocytes play essential roles in tissue modeling during embryogenesis and morphogenesis, and also in innate immunity. The open circulatory system of Drosophila makes haemocytes ideal signal mediators to cells and tissues in response to events such as infection and wounding. The application of recently developed and sophisticated genetic tools to the relatively simple genome of Drosophila has made the fly a popular system for modeling human tumorigensis and metastasis. Drosophila is now used for screening and investigation of genes implicated in human leukemia and also in modeling development of solid tumors. This second line of research offers promising opportunities to determine the seemingly conflicting roles of blood cells in tumor progression and invasion. This review provides an overview of the signaling pathways conserved in Drosophila during haematopoiesis, haemostasis, innate immunity, wound healing and inflammation. We also review the most recent progress in the use of Drosophila as a cancer research model with an emphasis on the roles haemocytes can play in various cancer models and in the links between inflammation and cancer. PMID:24409421

  12. Segmentation gene expression in the mothmidge Clogmia albipunctata (Diptera, psychodidae) and other primitive dipterans.

    PubMed

    Rohr, K B; Tautz, D; Sander, K

    1999-03-01

    To obtain a clearer understanding of the evolutionary transition between short- and long-germ modes of embryogenesis in insects, we studied the expression of two gap genes hunchback (hb) and Krüppel (Kr) as well as the pair-rule gene even-skipped (eve) in the dipteran Clogmia albipunctata (Nematocera, Psychodidae). This species has features of both short- and long-germ mode of embryogenesis. In Clogmia hb expression deviates from that known in Drosophila in two main respects: (1) it shows an extended dorsal domain that is linked to the large serosa anlage, and (2) it shows a terminal expression in the proctodeal region. These expression patterns are reminiscent of the hb expression pattern in the beetle Tribolium, which has a short germ mode of embryogenesis. Krüppel expression, on the other hand, was found to be rather similar to the Drosophila expression, both at early and late stages. eve expression starts with six stripes formed at blastoderm stage, while the seventh is only formed after the onset of gastrulation and germband extension. Surprisingly, no segmental secondary Eve stripes could be observed in Clogmia although such segmental stripes are known from higher dipterans, beetles and hymenopterans. We therefore also studied another nematoceran, Coboldia, to address this question and found that some segmental stripes form by intercalation as in Drosophila, although belatedly. Our results suggest that Clogmia embryogenesis, both with respect to morphological and molecular characteristics represents an intermediate between the long-germ mode known from higher dipterans such as Drosophila, and the short-germ mode found in more ancestral insects. PMID:10079357

  13. Resources for Biological Annotation of the Drosophila Genome

    SciTech Connect

    Gerald M. Rubin

    2005-08-08

    This project supported seed money for the development of cDNA and genetic resources to support studies of the Drosophila melanogaster genome. Key publications supported by this work that provide additional detail: (1) ''The Drosophila gene collection: identification of putative full-length cDNAs for 70% of D. melanogaster genes''; and (2) ''The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes''.

  14. Automatic brain tumor segmentation

    NASA Astrophysics Data System (ADS)

    Clark, Matthew C.; Hall, Lawrence O.; Goldgof, Dmitry B.; Velthuizen, Robert P.; Murtaugh, F. R.; Silbiger, Martin L.

    1998-06-01

    A system that automatically segments and labels complete glioblastoma-multiform tumor volumes in magnetic resonance images of the human brain is presented. The magnetic resonance images consist of three feature images (T1- weighted, proton density, T2-weighted) and are processed by a system which integrates knowledge-based techniques with multispectral analysis and is independent of a particular magnetic resonance scanning protocol. Initial segmentation is performed by an unsupervised clustering algorithm. The segmented image, along with cluster centers for each class are provided to a rule-based expert system which extracts the intra-cranial region. Multispectral histogram analysis separates suspected tumor from the rest of the intra-cranial region, with region analysis used in performing the final tumor labeling. This system has been trained on eleven volume data sets and tested on twenty-two unseen volume data sets acquired from a single magnetic resonance imaging system. The knowledge-based tumor segmentation was compared with radiologist-verified `ground truth' tumor volumes and results generated by a supervised fuzzy clustering algorithm. The results of this system generally correspond well to ground truth, both on a per slice basis and more importantly in tracking total tumor volume during treatment over time.

  15. [Toxic anterior segment syndrome].

    PubMed

    Cornut, P-L; Chiquet, C

    2011-01-01

    Toxic anterior segment syndrome (TASS) is a general term used to describe acute, sterile postoperative inflammation due to a non-infectious substance that accidentally enters the anterior segment at the time of surgery and mimics infectious endophthalmitis. TASS most commonly occurs acutely following anterior segment surgery, typically 12-72h after cataract extraction. Anterior segment inflammation is usually quite severe with hypopyon. Endothelial cell damage is common, resulting in diffuse corneal edema. No bacterium is isolated from ocular samples. The causes of TASS are numerous and difficult to isolate. Any device or substance used during the surgery or in the immediate postoperative period may be implicated. The major known causes include: preservatives in ophthalmic solutions, denatured ophthalmic viscosurgical devices, bacterial endotoxin, and intraocular lens-induced inflammation. Clinical features of infectious and non-infectious inflammation are initially indistinguishable and TASS is usually diagnosed and treated as acute endophthalmitis. It usually improves with local steroid treatment but may result in chronic elevation of intraocular pressure or irreversible corneal edema due to permanent damage of trabecular meshwork or endothelial cells. PMID:21176994

  16. Histone Gene Multiplicity and Position Effect Variegation in DROSOPHILA MELANOGASTER

    PubMed Central

    Moore, Gerald D.; Sinclair, Donald A.; Grigliatti, Thomas A.

    1983-01-01

    The histone genes of wild-type Drosophila melanogaster are reiterated 100–150 times per haploid genome and are located in the segment of chromosome 2 that corresponds to polytene bands 39D2-3 to E1-2. The influence of altered histone gene multiplicity on chromatin structure has been assayed by measuring modification of the gene inactivation associated with position effect variegation in genotypes bearing deletions of the 39D-E segment. The proportion of cells in which a variegating gene is active is increased in genotypes that are heterozygous for a deficiency that removes the histone gene complex. Deletions that remove segments adjacent to the histone gene complex have no effect on the expression of variegating genes. Suppression of position effect variegation associated with reduction of histone gene multiplicity applies to both X-linked and autosomal variegating genes. Position effects exerted by both autosomal and sex-chromosome heterochromatin were suppressible by deletions of the histone gene complex. The suppression was independent of the presence of the Y chromosome. A deficiency that deletes only the distal portion of the histone gene complex also has the ability to suppress position effect variegation. Duplication of the histone gene complex did not enhance position effect variegation. Deletion or duplication of the histone gene complex in the maternal genome had no effect on the extent of variegation in progeny whose histone gene multiplicity was normal. These results are discussed with respect to current knowledge of the organization of the histone gene complex and control of its expression. PMID:17246163

  17. Development of Drosophila larval neuromuscular junctions: maintaining synaptic strength.

    PubMed

    Li, H; Peng, X; Cooper, R L

    2002-01-01

    In spite of the available information about the development of Drosophila neuromuscular junctions, the correlation between nerve terminal morphology and maintenance of synaptic strength has still not been systematically addressed throughout larval development. We characterized the growth of the abdominal longitudinal muscle 6 (m6) and the motor terminals Ib and Is that innervate it within segment 4. In addition, we measured the evoked excitatory junction potential (EJP) amplitudes while the Ib and Is axons were selectively recruited. Regression analysis with natural log transformation of response variables indicated that the developmental curves for m6 and the motor axons Ib and Is were best fitted as second order polynomial regressions during larval development. Initially Is terminals are longer and possess more synaptic varicosities at the first instar stage. The Is terminals also grow faster in subsequent developmental stages. The growth of nerve terminals and their target m6 are not proportional although tightly correlated. This results in a larger average muscle area innervated by a single varicosity as the animal develops. The amplitudes of the EJPs of Ib and Is neurons show no developmental difference in their amplitudes from the first to the late third larval instar. The Is axon consistently produced larger EJPs than the Ib axon at each developmental stage. The time constants for both rising and decay phases of EJPs increase exponentially throughout larval development. The results presented not only help in quantifying the normal development of Drosophila neuromuscular junctions, but also provide a framework for future investigations to properly interpret developmental abnormalities that may occur in various mutants. PMID:12421617

  18. Enhancing Undergraduate Teaching and Research with a "Drosophila" Virginizing System

    ERIC Educational Resources Information Center

    Venema, Dennis R.

    2006-01-01

    Laboratory exercises using "Drosophila" crosses are an effective pedagogical method to complement traditional lecture and textbook presentations of genetics. Undergraduate thesis research is another common setting for using "Drosophila." A significant barrier to using "Drosophila" for undergraduate teaching or research is the time and skill…

  19. An integrated hybrid microfluidic device for oviposition-based chemical screening of adult Drosophila melanogaster.

    PubMed

    Leung, Jacob C K; Hilliker, Arthur J; Rezai, Pouya

    2016-02-21

    Chemical screening using Drosophila melanogaster (the fruit fly) is vital in drug discovery, agricultural, and toxicological applications. Oviposition (egg laying) on chemically-doped agar plates is an important read-out metric used to quantitatively assess the biological fitness and behavioral responses of Drosophila. Current oviposition-based chemical screening studies are inaccurate, labor-intensive, time-consuming, and inflexible due to the manual chemical doping of agar. In this paper, we have developed a novel hybrid agar-polydimethylsiloxane (PDMS) microfluidic device for single- and multi-concentration chemical dosing and on-chip oviposition screening of free-flying adult stage Drosophila. To achieve this, we have devised a novel technique to integrate agar with PDMS channels using ice as a sacrificial layer. Subsequently, we have conducted single-chemical toxicity and multiple choice chemical preference assays on adult Drosophila melanogaster using zinc and acetic acid at various concentrations. Our device has enabled us to 1) demonstrate that Drosophila is capable of sensing the concentration of different chemicals on a PDMS-agar microfluidic device, which plays significant roles in determining oviposition site selection and 2) investigate whether oviposition preference differs between single- and multi-concentration chemical environments. This device may be used to study fundamental and applied biological questions in Drosophila and other egg laying insects. It can also be extended in design to develop sophisticated and dynamic chemical dosing and high-throughput screening platforms in the future that are not easily achievable with the existing oviposition screening techniques. PMID:26768402

  20. The peripheral nervous system supports blood cell homing and survival in the Drosophila larva

    PubMed Central

    Makhijani, Kalpana; Alexander, Brandy; Tanaka, Tsubasa; Rulifson, Eric; Brückner, Katja

    2011-01-01

    Interactions of hematopoietic cells with their microenvironment control blood cell colonization, homing and hematopoiesis. Here, we introduce larval hematopoiesis as the first Drosophila model for hematopoietic colonization and the role of the peripheral nervous system (PNS) as a microenvironment in hematopoiesis. The Drosophila larval hematopoietic system is founded by differentiated hemocytes of the embryo, which colonize segmentally repeated epidermal-muscular pockets and proliferate in these locations. Importantly, we show that these resident hemocytes tightly colocalize with peripheral neurons and we demonstrate that larval hemocytes depend on the PNS as an attractive and trophic microenvironment. atonal (ato) mutant or genetically ablated larvae, which are deficient for subsets of peripheral neurons, show a progressive apoptotic decline in hemocytes and an incomplete resident hemocyte pattern, whereas supernumerary peripheral neurons induced by ectopic expression of the proneural gene scute (sc) misdirect hemocytes to these ectopic locations. This PNS-hematopoietic connection in Drosophila parallels the emerging role of the PNS in hematopoiesis and immune functions in vertebrates, and provides the basis for the systematic genetic dissection of the PNS-hematopoietic axis in the future. PMID:22071105

  1. Environmental ethanol as an ecological constraint on dietary breadth of Spotted-Wing Drosophila, Drosophila suzukii Mat. (Diptera: Drosophilidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spotted-wing Drosophila (SWD), Drosophila suzukii, is a recent fruit pest of the Americas whose destructiveness stems from its subcutaneous insertion of eggs into cultivated berries via a female’s prominent double bladed and serrated ovipositor. Atypical of most other Drosophila, D. suzukii adults a...

  2. 31 Flavors of Drosophila Rab proteins

    SciTech Connect

    Zhang, Jun; Schulze, Karen L.; Hiesinger, P. Robin; Suyama, Kaye; Wang, Stream; Fish, Matthew; Acar, Melih; Hoskins, Roger A.; Bellen, HugoJ.; Scott, Matthew P.

    2007-04-03

    Rab proteins are small GTPases that play important roles intransport of vesicle cargo and recruitment, association of motor andother proteins with vesicles, and docking and fusion of vesicles atdefined locations. In vertebrates, more than 75 Rab genes have beenidentified, some of which have been intensively studied for their rolesin endosome and synaptic vesicle trafficking. Recent studies of thefunctions of certain Rab proteins have revealed specific roles inmediating developmental signal transduction. We have begun a systematicgenetic study of the 33 Rab genes in Drosophila. Most of the fly proteinsare clearly related to specific vertebrate proteins. We report here thecreation of a set of transgenic fly lines that allow spatially andtemporally regulated expression of Drosophila Rab proteins. We generatedfluorescent protein-tagged wild-type, dominant-negative, andconstitutively active forms of 31 Drosophila Rab proteins. We describeDrosophila Rab expression patterns during embryogenesis, the subcellularlocalization of some Rab proteins, and comparisons of the localization ofwild-type, dominant-negative, and constitutively active forms of selectedRab proteins. The high evolutionary conservation and low redundancy ofDrosophila Rab proteins make these transgenic lines a useful toolkit forinvestigating Rab functions in vivo.

  3. Gut-associated microbes of Drosophila melanogaster

    PubMed Central

    Broderick, Nichole; Lemaitre, Bruno

    2012-01-01

    There is growing interest in using Drosophila melanogaster to elucidate mechanisms that underlie the complex relationships between a host and its microbiota. In addition to the many genetic resources and tools Drosophila provides, its associated microbiota is relatively simple (1–30 taxa), in contrast to the complex diversity associated with vertebrates (> 500 taxa). These attributes highlight the potential of this system to dissect the complex cellular and molecular interactions that occur between a host and its microbiota. In this review, we summarize what is known regarding the composition of gut-associated microbes of Drosophila and their impact on host physiology. We also discuss these interactions in the context of their natural history and ecology and describe some recent insights into mechanisms by which Drosophila and its gut microbiota interact. “Workers with Drosophila have been considered fortunate in that they deal with the first multicellular invertebrate to be cultured monoxenically (Delcourt and Guyenot, 1910); the first to be handled axenically on a semisynthetic diet (Guyenot, 1917); and the first to be grown on a defined diet (Schultz et al., 1946). This list of advantages is somewhat embarrassing, since it implies an interest in nutrition that, in reality, was only secondary. The very first studies were concerned with the reduction of variability in genetic experiments (Delcourt and Guyenot, 1910) and standardization of the nutritional environment.” -James Sang, 1959 Ann NY Acad 1 PMID:22572876

  4. Probability of rupture of multiple fault segments

    USGS Publications Warehouse

    Andrews, D.J.; Schwerer, E.

    2000-01-01

    Fault segments identified from geologic and historic evidence have sometimes been adopted as features limiting the likely extends of earthquake ruptures. There is no doubt that individual segments can sometimes join together to produce larger earthquakes. This work is a trial of an objective method to determine the probability of multisegment ruptures. The frequency of occurrence of events on all conjectured combinations of adjacent segments in northern California is found by fitting to both geologic slip rates and to an assumed distribution of event sizes for the region as a whole. Uncertainty in the shape of the distribution near the maximum magnitude has a large effect on the solution. Frequencies of individual events cannot be determined, but it is possible to find a set of frequencies to fit a model closely. A robust conclusion for the San Francisco Bay region is that large multisegment events occur on the San Andreas and San Gregorio faults, but single-segment events predominate on the extended Hayward and Calaveras strands of segments.

  5. Combined actions of multiple hairpin loop structures and sites of rate-limiting endonucleolytic cleavage determine differential degradation rates of individual segments within polycistronic puf operon mRNA.

    PubMed Central

    Klug, G; Cohen, S N

    1990-01-01

    Differential expression of the genes within the puf operon of Rhodobacter capsulatus is accomplished in part by differences in the rate of degradation of different segments of the puf transcript. We report here that decay of puf mRNA sequences specifying the light-harvesting I (LHI) and reaction center (RC) photosynthetic membrane peptides is initiated endoribonucleolytically within a discrete 1.4-kilobase segment of the RC-coding region. Deletion of this segment increased the half-life of the RC-coding region from 8 to 20 min while not affecting decay of LHI-coding sequences upstream from an intercistronic hairpin loop structure shown previously to impede 3'-to-5' degradation. Prolongation of RC segment half-life was dependent on the presence of other hairpin structures 3' to the RC region. Inserting the endonuclease-sensitive sites into the LHI-coding segment markedly accelerated its degradation. Our results suggest that differential degradation of the RC- and LHI-coding segments of puf mRNA is accomplished at least in part by the combined actions of RC region-specific endonuclease(s), one or more exonucleases, and several strategically located exonuclease-impeding hairpins. Images PMID:2394682

  6. Host Genetic Control of the Microbiota Mediates the Drosophila Nutritional Phenotype.

    PubMed

    Chaston, John M; Dobson, Adam J; Newell, Peter D; Douglas, Angela E

    2016-01-01

    A wealth of studies has demonstrated that resident microorganisms (microbiota) influence the pattern of nutrient allocation to animal protein and energy stores, but it is unclear how the effects of the microbiota interact with other determinants of animal nutrition, including animal genetic factors and diet. Here, we demonstrate that members of the gut microbiota in Drosophila melanogaster mediate the effect of certain animal genetic determinants on an important nutritional trait, triglyceride (lipid) content. Parallel analysis of the taxonomic composition of the associated bacterial community and host nutritional indices (glucose, glycogen, triglyceride, and protein contents) in multiple Drosophila genotypes revealed significant associations between the abundance of certain microbial taxa, especially Acetobacteraceae and Xanthamonadaceae, and host nutritional phenotype. By a genome-wide association study of Drosophila lines colonized with a defined microbiota, multiple host genes were statistically associated with the abundance of one bacterium, Acetobacter tropicalis. Experiments using mutant Drosophila validated the genetic association evidence and reveal that host genetic control of microbiota abundance affects the nutritional status of the flies. These data indicate that the abundance of the resident microbiota is influenced by host genotype, with consequent effects on nutrient allocation patterns, demonstrating that host genetic control of the microbiome contributes to the genotype-phenotype relationship of the animal host. PMID:26567306

  7. Host Genetic Control of the Microbiota Mediates the Drosophila Nutritional Phenotype

    PubMed Central

    Chaston, John M.; Dobson, Adam J.; Newell, Peter D.

    2015-01-01

    A wealth of studies has demonstrated that resident microorganisms (microbiota) influence the pattern of nutrient allocation to animal protein and energy stores, but it is unclear how the effects of the microbiota interact with other determinants of animal nutrition, including animal genetic factors and diet. Here, we demonstrate that members of the gut microbiota in Drosophila melanogaster mediate the effect of certain animal genetic determinants on an important nutritional trait, triglyceride (lipid) content. Parallel analysis of the taxonomic composition of the associated bacterial community and host nutritional indices (glucose, glycogen, triglyceride, and protein contents) in multiple Drosophila genotypes revealed significant associations between the abundance of certain microbial taxa, especially Acetobacteraceae and Xanthamonadaceae, and host nutritional phenotype. By a genome-wide association study of Drosophila lines colonized with a defined microbiota, multiple host genes were statistically associated with the abundance of one bacterium, Acetobacter tropicalis. Experiments using mutant Drosophila validated the genetic association evidence and reveal that host genetic control of microbiota abundance affects the nutritional status of the flies. These data indicate that the abundance of the resident microbiota is influenced by host genotype, with consequent effects on nutrient allocation patterns, demonstrating that host genetic control of the microbiome contributes to the genotype-phenotype relationship of the animal host. PMID:26567306

  8. A family of snail-related zinc finger proteins regulates two distinct and parallel mechanisms that mediate Drosophila neuroblast asymmetric divisions.

    PubMed

    Cai, Y; Chia, W; Yang, X

    2001-04-01

    Three snail family genes snail, escargot and worniu, encode related zinc finger transcription factors that mediate Drosophila central nervous system (CNS) development. We show that simultaneous removal of all three genes causes defective neuroblast asymmetric divisions; inscuteable transcription/translation is delayed/suppressed in the segmented CNS. Further more, defects in localization of cell fate determinants and orientation of the mitotic spindle in dividing neuroblasts are much stronger than those associated with inscuteable loss of function. In inscuteable neuroblasts, cell fate determinants are mislocalized during prophase and metaphase, yet during anaphase and telophase the great majority of mutant neuroblasts localize these determinants as cortical crescents overlying one of the spindle poles. This phenomenon, known as 'telophase rescue', does not occur in the absence of the snail family genes; moreover, in contrast to inscuteable mutants, mitotic spindle orientation is completely randomized. Our data provide further evidence for the existence of two distinct asymmetry-controlling mechanisms in neuroblasts both of which require snail family gene function: an inscuteable-dependent mechanism that functions throughout mitosis and an inscuteable-independent mechanism that acts during anaphase/telophase. PMID:11285234

  9. Globus pallidus internal segment.

    PubMed

    Nambu, Atsushi

    2007-01-01

    The internal segment of the globus pallidus (GP(i)) gathers many bits of information including movement-related activity from the striatum, external segment of the globus pallidus (GP(e)), and subthalamic nucleus (STN), and integrates them. The GP(i) receives rich GABAergic inputs from the striatum and GP(e), and gamma-aminobutyric acid (GABA) receptors are distributed in the GP(i) in a specific manner. Thus, inputs from the striatum and GP(e) may control GP(i) activity in a different way. The GP(i) finally conveys processed information outside the basal ganglia. Changes in GABAergic neurotransmission have been reported in movement disorders and suggested to play an important role in the pathophysiology of the symptoms. PMID:17499112

  10. Segmented vortex flaps

    NASA Technical Reports Server (NTRS)

    Rao, D. M.

    1983-01-01

    Segmented vortex flaps were suggested as a means of delaying the vortex spill-over causing thrust loss over the outboard region of single-panel flaps. Also proposed was hinge-line setback for exploiting leading-edge suction in conjunction with vortex flaps to improve the overall thrust per unit flap area. These two concepts in combination were tested on a 60-deg cropped delta wing model. Significant improvement in flap efficiency was indicated by a reduction of the flap/wing area from 11.4% of single-panel flap to 6.3% of a two segment delta flap design, with no lift/drag penalty at lift coefficients between 0.5 and 0.7. The more efficient vortex flap arrangement of this study should benefit the performance attainable with flaps of given area on wings of moderate leading-edge sweep.

  11. Segmented Thermal Barrier Coating

    NASA Technical Reports Server (NTRS)

    Smialek, James L. (Inventor)

    2001-01-01

    The article has a macro-segmented thermal barrier coating due to the presence of a pattern of three-dimensional features. The features may be a series of raised ribs formed on the substrate surface and being spaced from 0.05 inches to 0.30 apart. The ribs have a width ranging from 0.005 inches to 0.02 inches, and a height ranging from 25% to 100% of the thickness of the barrier coating. Alternately, the features may be a similar pattern of grooves formed in the surface of the substrate. Other embodiments provide segmentation by grooves or ribs in the bond coat or alternately grooves formed in the thermal barrier layer.

  12. Drosophila melanogaster prefers compounds perceived sweet by humans.

    PubMed

    Gordesky-Gold, Beth; Rivers, Natasha; Ahmed, Osama M; Breslin, Paul A S

    2008-03-01

    To understand the functional similarities of fly and mammalian taste receptors, we used a top-down approach that first established the fly sweetener-response profile. We employed the fruit fly Drosophila melanogaster, an omnivorous human commensal, and determined its sensitivity to an extended set of stimuli that humans find sweet. Flies were tested with all sweeteners in 2 assays that measured their taste reactivity (proboscis extension assay) and their ingestive preferences (free roaming ingestion choice test). A total of 21 sweeteners, comprised of 11 high-potency sweeteners, 2 amino acids, 5 sugars, 2 sugar alcohols, and a sweet salt (PbCl2), were tested in both assays. We found that wild-type Drosophila responded appetitively to most high-potency sweeteners preferred by humans, even those not considered sweet by rodents or new world monkeys. The similarities in taste preferences for sweeteners suggest that frugivorous/omnivorous apes and flies have evolved promiscuous carbohydrate taste detectors with similar affinities for myriad high-potency sweeteners. Whether these perceptual parallels are the result of convergent evolution of saccharide receptor-binding mechanisms remains to be determined. PMID:18234713

  13. Chromosomal localization of microsatellite loci in Drosophila mediopunctata.

    PubMed

    Cavasini, Renato; Batista, Marcos Roberto Dias; Klaczko, Louis Bernard

    2015-03-01

    Drosophila mediopunctata has been used as a model organism for genetics and evolutionary studies in the last three decades. A linkage map with 48 microsatellite loci recently published for this species showed five syntenic groups, which had their homology determined to Drosophila melanogaster chromosomes. Then, by inference, each of the groups was associated with one of the five major chromosomes of D. mediopunctata. Our objective was to carry out a genetic (chromosomal) analysis to increase the number of available loci with known chromosomal location. We made a simultaneous analysis of visible mutant phenotypes and microsatellite genotypes in a backcross of a standard strain and a mutant strain, which had each major autosome marked. Hence, we could establish the chromosomal location of seventeen loci; including one from each of the five major linkage groups previously published, and twelve new loci. Our results were congruent with the previous location and they open new possibilities to future work integrating microsatellites, chromosomal inversions, and genetic determinants of physiological and morphological variation. PMID:25983625

  14. The behaviour of Drosophila adult hindgut stem cells is controlled by Wnt and Hh signalling.

    PubMed

    Takashima, Shigeo; Mkrtchyan, Marianna; Younossi-Hartenstein, Amelia; Merriam, John R; Hartenstein, Volker

    2008-07-31

    The intestinal tract maintains proper function by replacing aged cells with freshly produced cells that arise from a population of self-renewing intestinal stem cells (ISCs). In the mammalian intestine, ISC self renewal, amplification and differentiation take place along the crypt-villus axis, and are controlled by the Wnt and hedgehog (Hh) signalling pathways. However, little is known about the mechanisms that specify ISCs within the developing intestinal epithelium, or about the signalling centres that help maintain them in their self-renewing stem cell state. Here we show that in adult Drosophila melanogaster, ISCs of the posterior intestine (hindgut) are confined to an anterior narrow segment, which we name the hindgut proliferation zone (HPZ). Within the HPZ, self renewal of ISCs, as well as subsequent proliferation and differentiation of ISC descendants, are controlled by locally emanating Wingless (Wg, a Drosophila Wnt homologue) and Hh signals. The anteriorly restricted expression of Wg in the HPZ acts as a niche signal that maintains cells in a slow-cycling, self-renewing mode. As cells divide and move posteriorly away from the Wg source, they enter a phase of rapid proliferation. During this phase, Hh signal is required for exiting the cell cycle and the onset of differentiation. The HPZ, with its characteristic proliferation dynamics and signalling properties, is set up during the embryonic phase and becomes active in the larva, where it generates all adult hindgut cells including ISCs. The mechanism and genetic control of cell renewal in the Drosophila HPZ exhibits a large degree of similarity with what is seen in the mammalian intestine. Our analysis of the Drosophila HPZ provides an insight into the specification and control of stem cells, highlighting the way in which the spatial pattern of signals that promote self renewal, growth and differentiation is set up within a genetically tractable model system. PMID:18633350

  15. Probing Polymer-Segment Motions By ESR

    NASA Technical Reports Server (NTRS)

    Tsay, Fun-Dow; Gupta, Amitava

    1988-01-01

    Molecular origins of mechanical properties and aging processes studied. Rotational motions of segments of poly(methyl methacrylate) molecules studied theoretically and experimentally. Activation energies of these motions as determined from temperature dependencies of ESR spectra agree closely with predictions of theory.

  16. Example based lesion segmentation

    NASA Astrophysics Data System (ADS)

    Roy, Snehashis; He, Qing; Carass, Aaron; Jog, Amod; Cuzzocreo, Jennifer L.; Reich, Daniel S.; Prince, Jerry; Pham, Dzung

    2014-03-01

    Automatic and accurate detection of white matter lesions is a significant step toward understanding the progression of many diseases, like Alzheimer's disease or multiple sclerosis. Multi-modal MR images are often used to segment T2 white matter lesions that can represent regions of demyelination or ischemia. Some automated lesion segmentation methods describe the lesion intensities using generative models, and then classify the lesions with some combination of heuristics and cost minimization. In contrast, we propose a patch-based method, in which lesions are found using examples from an atlas containing multi-modal MR images and corresponding manual delineations of lesions. Patches from subject MR images are matched to patches from the atlas and lesion memberships are found based on patch similarity weights. We experiment on 43 subjects with MS, whose scans show various levels of lesion-load. We demonstrate significant improvement in Dice coefficient and total lesion volume compared to a state of the art model-based lesion segmentation method, indicating more accurate delineation of lesions.

  17. Apoptosis in Drosophila: which role for mitochondria?

    PubMed

    Clavier, Amandine; Rincheval-Arnold, Aurore; Colin, Jessie; Mignotte, Bernard; Guénal, Isabelle

    2016-03-01

    It is now well established that the mitochondrion is a central regulator of mammalian cell apoptosis. However, the importance of this organelle in non-mammalian apoptosis has long been regarded as minor, mainly because of the absence of a crucial role for cytochrome c in caspase activation. Recent results indicate that the control of caspase activation and cell death in Drosophila occurs at the mitochondrial level. Numerous proteins, including RHG proteins and proteins of the Bcl-2 family that are key regulators of Drosophila apoptosis, constitutively or transiently localize in mitochondria. These proteins participate in the cell death process at different levels such as degradation of Diap1, a Drosophila IAP, production of mitochondrial reactive oxygen species or stimulation of the mitochondrial fission machinery. Here, we review these mitochondrial events that might have their counterpart in human. PMID:26679112

  18. The digestive tract of Drosophila melanogaster.

    PubMed

    Lemaitre, Bruno; Miguel-Aliaga, Irene

    2013-01-01

    The digestive tract plays a central role in the digestion and absorption of nutrients. Far from being a passive tube, it provides the first line of defense against pathogens and maintains energy homeostasis by exchanging neuronal and endocrine signals with other organs. Historically neglected, the gut of the fruit fly Drosophila melanogaster has recently come to the forefront of Drosophila research. Areas as diverse as stem cell biology, neurobiology, metabolism, and immunity are benefitting from the ability to study the genetics of development, growth regulation, and physiology in the same organ. In this review, we summarize our knowledge of the Drosophila digestive tract, with an emphasis on the adult midgut and its functional underpinnings. PMID:24016187

  19. Viruses and Antiviral Immunity in Drosophila

    PubMed Central

    Xu, Jie; Cherry, Sara

    2013-01-01

    Viral pathogens present many challenges to organisms, driving the evolution of a myriad of antiviral strategies to combat infections. A wide variety of viruses infect invertebrates, including both natural pathogens that are insect-restricted, and viruses that are transmitted to vertebrates. Studies using the powerful tools available in the model organism Drosophila have expanded our understanding of antiviral defenses against diverse viruses. In this review, we will cover three major areas. First, we will describe the tools used to study viruses in Drosophila. Second, we will survey the major viruses that have been studied in Drosophila. And lastly, we will discuss the well-characterized mechanisms that are active against these diverse pathogens, focusing on non-RNAi mediated antiviral mechanisms. Antiviral RNAi is discussed in another paper in this issue. PMID:23680639

  20. Receptor Tyrosine Kinases in Drosophila Development

    PubMed Central

    Sopko, Richelle; Perrimon, Norbert

    2013-01-01

    Tyrosine phosphorylation plays a significant role in a wide range of cellular processes. The Drosophila genome encodes more than 20 receptor tyrosine kinases and extensive studies in the past 20 years have illustrated their diverse roles and complex signaling mechanisms. Although some receptor tyrosine kinases have highly specific functions, others strikingly are used in rather ubiquitous manners. Receptor tyrosine kinases regulate a broad expanse of processes, ranging from cell survival and proliferation to differentiation and patterning. Remarkably, different receptor tyrosine kinases share many of the same effectors and their hierarchical organization is retained in disparate biological contexts. In this comprehensive review, we summarize what is known regarding each receptor tyrosine kinase during Drosophila development. Astonishingly, very little is known for approximately half of all Drosophila receptor tyrosine kinases. PMID:23732470

  1. Developmental Toxicity Assays Using the Drosophila Model

    PubMed Central

    Rand, Matthew D.; Montgomery, Sara L.; Prince, Lisa; Vorojeikina, Daria

    2014-01-01

    The fruit fly (Drosophila melanogaster) has long been a premier model for developmental biologists and geneticists. The utility of Drosophila for toxicology studies has only recently gained broader recognition as a tool to elaborate molecular genetic mechanisms of toxic substances. In this article two practical applications of Drosophila for developmental toxicity assays are described. The first assay takes advantage of newly developed methods to render the fly embryo accessible to small molecules, toxicants and drugs. The second assay engages straightforward exposures to developing larvae and easy to score outcomes of adult development. With the extensive collections of flies that are publicly available and the ease with which to create transgenic flies, these two assays have a unique power for identifying and characterizing molecular mechanisms and cellular pathways specific to the mode of action of a number of toxicants and drugs. PMID:24789363

  2. Live cell imaging in Drosophila melanogaster.

    PubMed

    Parton, Richard M; Vallés, Ana Maria; Dobbie, Ian M; Davis, Ilan

    2010-04-01

    Although many of the techniques of live cell imaging in Drosophila melanogaster are also used by the greater community of cell biologists working on other model systems, studying living fly tissues presents unique difficulties with regard to keeping the cells alive, introducing fluorescent probes, and imaging through thick, hazy cytoplasm. This article outlines the major tissue types amenable to study by time-lapse cinematography and different methods for keeping the cells alive. It describes various imaging and associated techniques best suited to following changes in the distribution of fluorescently labeled molecules in real time in these tissues. Imaging, in general, is a rapidly developing discipline, and recent advances in imaging technology are able to greatly extend what can be achieved with live cell imaging of Drosophila tissues. As far as possible, this article includes the latest technical developments and discusses likely future developments in imaging methods that could have an impact on research using Drosophila. PMID:20360379

  3. Volatile pheromone signalling in Drosophila.

    PubMed

    Smith, Dean P

    2012-03-01

    Once captured by the antenna, 11-cis vaccenyl acetate (cVA) binds to an extracellular binding protein called LUSH that undergoes a conformational shift upon cVA binding. The stable LUSH-cVA complex is the activating ligand for pheromone receptors present on the dendrites of the aT1 neurones, comprising the only neurones that detect cVA pheromone. This mechanism explains the single molecule sensitivity of insect pheromone detection systems. The receptor that recognizes activated LUSH consists of a complex of several proteins, including Or67d, a member of the tuning odourant receptor family, Orco, a co-receptor ion channel, and SNMP, a CD36 homologue that may be an inhibitory subunit. In addition, genetic screens and reconstitution experiments reveal additional factors that are important for pheromone detection. Identification and functional dissection of these factors in Drosophila melanogaster Meigen should permit the identification of homologous factors in pathogenic insects and agricultural pests, which, in turn, may be viable candidates for novel classes of compounds to control populations of target insect species without impacting beneficial species. PMID:24347807

  4. Automatic segmentation of clinical texts.

    PubMed

    Apostolova, Emilia; Channin, David S; Demner-Fushman, Dina; Furst, Jacob; Lytinen, Steven; Raicu, Daniela

    2009-01-01

    Clinical narratives, such as radiology and pathology reports, are commonly available in electronic form. However, they are also commonly entered and stored as free text. Knowledge of the structure of clinical narratives is necessary for enhancing the productivity of healthcare departments and facilitating research. This study attempts to automatically segment medical reports into semantic sections. Our goal is to develop a robust and scalable medical report segmentation system requiring minimum user input for efficient retrieval and extraction of information from free-text clinical narratives. Hand-crafted rules were used to automatically identify a high-confidence training set. This automatically created training dataset was later used to develop metrics and an algorithm that determines the semantic structure of the medical reports. A word-vector cosine similarity metric combined with several heuristics was used to classify each report sentence into one of several pre-defined semantic sections. This baseline algorithm achieved 79% accuracy. A Support Vector Machine (SVM) classifier trained on additional formatting and contextual features was able to achieve 90% accuracy. Plans for future work include developing a configurable system that could accommodate various medical report formatting and content standards. PMID:19965054

  5. Recombining without Hotspots: A Comprehensive Evolutionary Portrait of Recombination in Two Closely Related Species of Drosophila

    PubMed Central

    Smukowski Heil, Caiti S.; Ellison, Chris; Dubin, Matthew; Noor, Mohamed A.F.

    2015-01-01

    Meiotic recombination rate varies across the genome within and between individuals, populations, and species in virtually all taxa studied. In almost every species, this variation takes the form of discrete recombination hotspots, determined in some mammals by a protein called PRDM9. Hotspots and their determinants have a profound effect on the genomic landscape, and share certain features that extend across the tree of life. Drosophila, in contrast, are anomalous in their absence of hotspots, PRDM9, and other species-specific differences in the determination of recombination. To better understand the evolution of meiosis and general patterns of recombination across diverse taxa, we present a truly comprehensive portrait of recombination across time, combining recently published cross-based contemporary recombination estimates from each of two sister species with newly obtained linkage-disequilibrium-based historic estimates of recombination from both of these species. Using Drosophila pseudoobscura and Drosophila miranda as a model system, we compare recombination rate between species at multiple scales, and we suggest that Drosophila replicate the pattern seen in human–chimpanzee in which recombination rate is conserved at broad scales. We also find evidence of a species-wide recombination modifier(s), resulting in both a present and historic genome-wide elevation of recombination rates in D. miranda, and identify broad scale effects on recombination from the presence of an inversion. Finally, we reveal an unprecedented view of the distribution of recombination in D. pseudoobscura, illustrating patterns of linked selection and where recombination is taking place. Overall, by combining these estimation approaches, we highlight key similarities and differences in recombination between Drosophila and other organisms. PMID:26430062

  6. Mutagenic effect of tritium on DNA of Drosophila melanogaster. Comprehensive performance report, December 15, 1985-June 1, 1988

    SciTech Connect

    Lee, W.R.

    1988-01-01

    The results of the RBE determination of tritium to cobalt-69 gamma radiation along with a description of methods of treatment and dose determination are given. Using the described procedures for exposing Drosophila to tritiated water, the authors induced mutations by tritium beta radiation and recovered them at the Adh locus.

  7. Segmenting the Adult Education Market.

    ERIC Educational Resources Information Center

    Aurand, Tim

    1994-01-01

    Describes market segmentation and how the principles of segmentation can be applied to the adult education market. Indicates that applying segmentation techniques to adult education programs results in programs that are educationally and financially satisfying and serve an appropriate population. (JOW)

  8. Market Segmentation for Information Services.

    ERIC Educational Resources Information Center

    Halperin, Michael

    1981-01-01

    Discusses the advantages and limitations of market segmentation as strategy for the marketing of information services made available by nonprofit organizations, particularly libraries. Market segmentation is defined, a market grid for libraries is described, and the segmentation of information services is outlined. A 16-item reference list is…

  9. Functional Segments in Tongue Movement

    ERIC Educational Resources Information Center

    Stone, Maureen; Epstein, Melissa A.; Iskarous, Khalil

    2004-01-01

    The tongue is a deformable object, and moves by compressing or expanding local functional segments. For any single phoneme, these functional tongue segments may move in similar or opposite directions, and may reach target maximum synchronously or not. This paper will discuss the independence of five proposed segments in the production of speech.…

  10. Evaluation of Traditional Medicines for Neurodegenerative Diseases Using Drosophila Models

    PubMed Central

    Lee, Soojin; Bang, Se Min; Lee, Joon Woo; Cho, Kyoung Sang

    2014-01-01

    Drosophila is one of the oldest and most powerful genetic models and has led to novel insights into a variety of biological processes. Recently, Drosophila has emerged as a model system to study human diseases, including several important neurodegenerative diseases. Because of the genomic similarity between Drosophila and humans, Drosophila neurodegenerative disease models exhibit a variety of human-disease-like phenotypes, facilitating fast and cost-effective in vivo genetic modifier screening and drug evaluation. Using these models, many disease-associated genetic factors have been identified, leading to the identification of compelling drug candidates. Recently, the safety and efficacy of traditional medicines for human diseases have been evaluated in various animal disease models. Despite the advantages of the Drosophila model, its usage in the evaluation of traditional medicines is only nascent. Here, we introduce the Drosophila model for neurodegenerative diseases and some examples demonstrating the successful application of Drosophila models in the evaluation of traditional medicines. PMID:24790636

  11. Drosophila Hook-Related Protein (Girdin) Is Essential for Sensory Dendrite Formation

    PubMed Central

    Ha, Andrew; Polyanovsky, Andrey; Avidor-Reiss, Tomer

    2015-01-01

    The dendrite of the sensory neuron is surrounded by support cells and is composed of two specialized compartments: the inner segment and the sensory cilium. How the sensory dendrite is formed and maintained is not well understood. Hook-related proteins (HkRP) like Girdin, DAPLE, and Gipie are actin-binding proteins, implicated in actin organization and in cell motility. Here, we show that the Drosophila melanogaster single member of the Hook-related protein family, Girdin, is essential for sensory dendrite formation and function. Mutations in girdin were identified during a screen for fly mutants with no mechanosensory function. Physiological, morphological, and ultrastructural studies of girdin mutant flies indicate that the mechanosensory neurons innervating external sensory organs (bristles) initially form a ciliated dendrite that degenerates shortly after, followed by the clustering of their cell bodies. Importantly, we observed that Girdin is expressed transiently during dendrite morphogenesis in three previously unidentified actin-based structures surrounding the inner segment tip and the sensory cilium. These actin structures are largely missing in girdin mutant. Defects in cilia are observed in other sensory organs such as those mediating olfaction and taste, suggesting that Girdin has a general role in forming sensory dendrites in Drosophila. These suggest that Girdin functions temporarily within the sensory organ and that this function is essential for the formation of the sensory dendrites via actin structures. PMID:26058848

  12. Rosa damascena decreased mortality in adult Drosophila.

    PubMed

    Jafari, Mahtab; Zarban, Asghar; Pham, Steven; Wang, Thomas

    2008-03-01

    The effects of a rose-flower extract, Rosa damascena, on the mortality rate of Drosophila melanogaster was evaluated in this study. R. damascena is a potent antioxidant that has many therapeutic uses in addition to its perfuming effects. Supplementing Drosophila with this rose extract resulted in a statistically significant decrease in mortality rate in male and female flies. Moreover, the observed anti-aging effects were not associated with common confounds of anti-aging properties, such as a decrease in fecundity or metabolic rate. PMID:18361732

  13. Neural control of aggression in Drosophila.

    PubMed

    Hoopfer, Eric D

    2016-06-01

    Like most animal species, fruit flies fight to obtain and defend resources essential to survival and reproduction. Aggressive behavior in Drosophila is genetically specified and also strongly influenced by the fly's social context, past experiences and internal states, making it an excellent framework for investigating the neural mechanisms that regulate complex social behaviors. Here, I summarize our current knowledge of the neural control of aggression in Drosophila and discuss recent advances in understanding the sensory pathways that influence the decision to fight or court, the neuromodulatory control of aggression, the neural basis by which internal states can influence both fighting and courtship, and how social experience modifies aggressive behavior. PMID:27179788

  14. The secret lives of Drosophila flies.

    PubMed

    Markow, Therese Ann

    2015-01-01

    Flies of the genus Drosophila, and particularly those of the species Drosophila melanogaster, are best known as laboratory organisms. As with all model organisms, they were domesticated for empirical studies, but they also continue to exist as wild populations. Decades of research on these flies in the laboratory have produced astounding and important insights into basic biological processes, but we have only scratched the surface of what they have to offer as research organisms. An outstanding challenge now is to build on this knowledge and explore how natural history has shaped D. melanogaster in order to advance our understanding of biology more generally. PMID:26041333

  15. Approach to nonparametric cooperative multiband segmentation with adaptive threshold.

    PubMed

    Sebari, Imane; He, Dong-Chen

    2009-07-10

    We present a new nonparametric cooperative approach to multiband image segmentation. It is based on cooperation between region-growing segmentation and edge segmentation. This approach requires no input data other than the images to be processed. It uses a spectral homogeneity criterion whose threshold is determined automatically. The threshold is adaptive and varies depending on the objects to be segmented. Applying this new approach to very high resolution satellite imagery has yielded satisfactory results. The approach demonstrated its performance on images of varied complexity and was able to detect objects of great spatial and spectral heterogeneity. PMID:19593349

  16. Mass segmentation using a combined method for cancer detection

    PubMed Central

    2011-01-01

    Background Breast cancer is one of the leading causes of cancer death for women all over the world and mammography is thought of as one of the main tools for early detection of breast cancer. In order to detect the breast cancer, computer aided technology has been introduced. In computer aided cancer detection, the detection and segmentation of mass are very important. The shape of mass can be used as one of the factors to determine whether the mass is malignant or benign. However, many of the current methods are semi-automatic. In this paper, we investigate fully automatic segmentation method. Results In this paper, a new mass segmentation algorithm is proposed. In the proposed algorithm, a fully automatic marker-controlled watershed transform is proposed to segment the mass region roughly, and then a level set is used to refine the segmentation. For over-segmentation caused by watershed, we also investigated different noise reduction technologies. Images from DDSM were used in the experiments and the results show that the new algorithm can improve the accuracy of mass segmentation. Conclusions The new algorithm combines the advantages of both methods. The combination of the watershed based segmentation and level set method can improve the efficiency of the segmentation. Besides, the introduction of noise reduction technologies can reduce over-segmentation. PMID:22784625

  17. The Minimal Word Hypothesis: A Speech Segmentation Strategy

    ERIC Educational Resources Information Center

    Meador, Diane L.

    1996-01-01

    Previous investigations have sought to determine how listeners might locate word boundaries in the speech signal for the purpose of lexical access. Cutler (1990) proposes the Metrical Segmentation Strategy (MSS), such that only full vowels in stressed syllables and their preceding syllabic onsets are segmented from the speech stream. I report the…

  18. Segmented nanowires displaying locally controllable properties

    DOEpatents

    Sutter, Eli Anguelova; Sutter, Peter Werner

    2013-03-05

    Vapor-liquid-solid growth of nanowires is tailored to achieve complex one-dimensional material geometries using phase diagrams determined for nanoscale materials. Segmented one-dimensional nanowires having constant composition display locally variable electronic band structures that are determined by the diameter of the nanowires. The unique electrical and optical properties of the segmented nanowires are exploited to form electronic and optoelectronic devices. Using gold-germanium as a model system, in situ transmission electron microscopy establishes, for nanometer-sized Au--Ge alloy drops at the tips of Ge nanowires (NWs), the parts of the phase diagram that determine their temperature-dependent equilibrium composition. The nanoscale phase diagram is then used to determine the exchange of material between the NW and the drop. The phase diagram for the nanoscale drop deviates significantly from that of the bulk alloy.

  19. Sorting of influenza A virus RNA genome segments after nuclear export

    SciTech Connect

    Takizawa, Naoki; Kumakura, Michiko; Takeuchi, Kaoru; Kobayashi, Nobuyuki; Nagata, Kyosuke

    2010-06-05

    The genome of the influenza A virus consists of eight different segments. These eight segments are thought to be sorted selectively in infected cells. However, the cellular compartment where segments are sorted is not known. We examined using temperature sensitive (ts) mutant viruses and cell fusion where segments are sorted in infected cells. Different cells were infected with different ts mutant viruses, and these cells were fused. In fused cells, genome segments are mixed only in the cytoplasm, because M1 prevents their re-import into the nucleus. We made a marker ts53 virus, which has silent mutations in given segments and determined the reassortment frequency on all segments using ts1 and marker ts53. In both co-infected and fused cells, all of marker ts53 segments and ts1 segments were incorporated into progeny virions in a random fashion. These results suggest that influenza virus genome segments are sorted after nuclear export.

  20. Accessory subunit of mitochondrial DNA polymerase from Drosophila embryos. Cloning, molecular analysis, and association in the native enzyme.

    PubMed

    Wang, Y; Farr, C L; Kaguni, L S

    1997-05-23

    A full-length cDNA of the accessory (beta) subunit of mitochondrial DNA polymerase from Drosophila embryos has been obtained, and its nucleotide sequence was determined. The cDNA clone encodes a polypeptide with a deduced amino acid sequence of 361 residues and a predicted molecular mass of 41 kDa. The gene encoding the beta subunit lies within 4 kilobase pairs of that for the catalytic subunit in the Drosophila genome, on the left arm of chromosome 2. The two genes have similar structural features and share several common DNA sequence elements in their upstream regions, suggesting the possibility of coordinate regulation. A human cDNA homolog of the accessory subunit was identified, and its nucleotide sequence was determined. The human sequence encodes a polypeptide with a predicted molecular mass of 43 kDa that shows a high degree of amino acid sequence similarity to the Drosophila beta subunit. Subunit-specific rabbit antisera, directed against the recombinant catalytic and accessory subunit polypeptides overexpressed and purified from Escherichia coli, recognize specifically and immunoprecipitate the native enzyme from Drosophila embryos. Demonstration of the physical association of the two subunits in the Drosophila enzyme and identification of a human accessory subunit homolog provide evidence for a common heterodimeric structure for animal mitochondrial DNA polymerases. PMID:9153213

  1. Interactive explorations of hierarchical segmentations

    NASA Technical Reports Server (NTRS)

    Tilton, James C.

    1992-01-01

    The authors report on the implementation of an interactive tool, called HSEGEXP, to interactively explore the hierarchical segmentation produced by the iterative parallel region growing (IPRG) algorithm to select the best segmentation result. This combination of the HSEGEXP tool with the IPRG algorithm amounts to a computer-assisted image segmentation system guided by human interaction. The initial application of the HSEGEXP tool is in the refinement of ground reference data based on the IPRG/HSEGEXP segmentation of the corresponding remotely sensed image data. The HSEGEXP tool is being used to help evaluate the effectiveness of an automatic 'best' segmentation process under development.

  2. Mutation of TweedleD, a member of an unconventional cuticle protein family, alters body shape in Drosophila

    PubMed Central

    Guan, Xiao; Middlebrooks, Brooke W.; Alexander, Sherry; Wasserman, Steven A.

    2006-01-01

    Body shape determination represents a critical aspect of morphogenesis. In the course of investigating body shape regulation in Drosophila, we have identified a dominant mutation, TweedleD1 (TwdlD1), that alters overall dimensions at the larval and pupal stages. Characterization of the affected locus led to the discovery of a gene family that has 27 members in Drosophila and is found only among insects. Analysis of gene expression at the RNA and protein levels revealed gene-specific temporal and spatial patterns in ectodermally derived tissues. In addition, light microscopic studies of fluorescently tagged proteins demonstrated that Tweedle proteins are incorporated into larval cuticular structures. This demonstration that a mutation in a Drosophila cuticular protein gene alters overall morphology confirms a role for the fly exoskeleton in determining body shape. Furthermore, parallels between these findings and studies of cuticle collagen genes in Caenorhabditis elegans suggest that the exoskeleton influences body shape in diverse organisms. PMID:17075064

  3. Assessment of the Effect of Intestinal Permeability Probes (Lactulose And Mannitol) and Other Liquids on Digesta Residence Times in Various Segments of the Gut Determined by Wireless Motility Capsule: A Randomised Controlled Trial

    PubMed Central

    Sequeira, Ivana R.; Lentle, Roger G.; Kruger, Marlena C.; Hurst, Roger D.

    2015-01-01

    Background Whilst the use of the mannitol/lactulose test for intestinal permeability has been long established it is not known whether the doses of these sugars modify transit time Similarly it is not known whether substances such as aspirin that are known to increase intestinal permeability to lactulose and mannitol and those such as ascorbic acid which are stated to be beneficial to gastrointestinal health also influence intestinal transit time. Methods Gastric and intestinal transit times were determined with a SmartPill following consumption of either a lactulose mannitol solution, a solution containing 600 mg aspirin, a solution containing 500 mg of ascorbic acid or an extract of blackcurrant, and compared by doubly repeated measures ANOVA with those following consumption of the same volume of a control in a cross-over study in six healthy female volunteers. The dominant frequencies of cyclic variations in gastric pressure recorded by the Smartpill were determined by fast Fourier transforms. Results The gastric transit times of lactulose mannitol solutions, of aspirin solutions and of blackcurrant juice did not differ from those of the control. The gastric transit times of the ascorbic acid solutions were significantly shorter than those of the other solutions. There were no significant differences between the various solutions either in the total small intestinal or colonic transit times. The intraluminal pHs during the initial quartiles of the small intestinal transit times were lower than those in the succeeding quartiles. This pattern did not vary with the solution that was consumed. The power of the frequencies of cyclic variation in intragastric pressure recorded by the Smartpill declined exponentially with increase in frequency and did not peak at the reported physiological frequencies of gastric contractile activity. Conclusions Whilst the segmental residence times were broadly similar to those using other methods, the high degree of variation between

  4. Arabic handwritten: pre-processing and segmentation

    NASA Astrophysics Data System (ADS)

    Maliki, Makki; Jassim, Sabah; Al-Jawad, Naseer; Sellahewa, Harin

    2012-06-01

    This paper is concerned with pre-processing and segmentation tasks that influence the performance of Optical Character Recognition (OCR) systems and handwritten/printed text recognition. In Arabic, these tasks are adversely effected by the fact that many words are made up of sub-words, with many sub-words there associated one or more diacritics that are not connected to the sub-word's body; there could be multiple instances of sub-words overlap. To overcome these problems we investigate and develop segmentation techniques that first segment a document into sub-words, link the diacritics with their sub-words, and removes possible overlapping between words and sub-words. We shall also investigate two approaches for pre-processing tasks to estimate sub-words baseline, and to determine parameters that yield appropriate slope correction, slant removal. We shall investigate the use of linear regression on sub-words pixels to determine their central x and y coordinates, as well as their high density part. We also develop a new incremental rotation procedure to be performed on sub-words that determines the best rotation angle needed to realign baselines. We shall demonstrate the benefits of these proposals by conducting extensive experiments on publicly available databases and in-house created databases. These algorithms help improve character segmentation accuracy by transforming handwritten Arabic text into a form that could benefit from analysis of printed text.

  5. Segmentation assisted food classification for dietary assessment

    NASA Astrophysics Data System (ADS)

    Zhu, Fengqing; Bosch, Marc; Schap, TusaRebecca; Khanna, Nitin; Ebert, David S.; Boushey, Carol J.; Delp, Edward J.

    2011-03-01

    Accurate methods and tools to assess food and nutrient intake are essential for the association between diet and health. Preliminary studies have indicated that the use of a mobile device with a built-in camera to obtain images of the food consumed may provide a less burdensome and more accurate method for dietary assessment. We are developing methods to identify food items using a single image acquired from the mobile device. Our goal is to automatically determine the regions in an image where a particular food is located (segmentation) and correctly identify the food type based on its features (classification or food labeling). Images of foods are segmented using Normalized Cuts based on intensity and color. Color and texture features are extracted from each segmented food region. Classification decisions for each segmented region are made using support vector machine methods. The segmentation of each food region is refined based on feedback from the output of classifier to provide more accurate estimation of the quantity of food consumed.

  6. Pulmonary lobe segmentation with level sets

    NASA Astrophysics Data System (ADS)

    Schmidt-Richberg, Alexander; Ehrhardt, Jan; Wilms, Matthias; Werner, René; Handels, Heinz

    2012-02-01

    Automatic segmentation of the separate human lung lobes is a crucial task in computer aided diagnostics and intervention planning, and required for example for determination of disease spreading or pulmonary parenchyma quantification. In this work, a novel approach for lobe segmentation based on multi-region level sets is presented. In a first step, interlobular fissures are detected using a supervised enhancement filter. The fissures are then used to compute a cost image, which is incorporated in the level set approach. By this, the segmentation is drawn to the fissures at places where structure information is present in the image. In areas with incomplete fissures (e.g. due to insufficient image quality or anatomical conditions) the smoothing term of the level sets applies and a closed continuation of the fissures is provided. The approach is tested on nine pulmonary CT scans. It is shown that incorporating the additional force term improves the segmentation significantly. On average, 83% of the left fissure is traced correctly; the right oblique and horizontal fissures are properly segmented to 76% and 48%, respectively.

  7. Segmentation Assisted Food Classification for Dietary Assessment.

    PubMed

    Zhu, Fengqing; Bosch, Marc; Schap, Tusarebecca; Khanna, Nitin; Ebert, David S; Boushey, Carol J; Delp, Edward J

    2011-01-24

    Accurate methods and tools to assess food and nutrient intake are essential for the association between diet and health. Preliminary studies have indicated that the use of a mobile device with a built-in camera to obtain images of the food consumed may provide a less burdensome and more accurate method for dietary assessment. We are developing methods to identify food items using a single image acquired from the mobile device. Our goal is to automatically determine the regions in an image where a particular food is located (segmentation) and correctly identify the food type based on its features (classification or food labeling). Images of foods are segmented using Normalized Cuts based on intensity and color. Color and texture features are extracted from each segmented food region. Classification decisions for each segmented region are made using support vector machine methods. The segmentation of each food region is refined based on feedback from the output of classifier to provide more accurate estimation of the quantity of food consumed. PMID:22128304

  8. Structure of the voltage-dependent potassium channel is highly conserved from Drosophila to vertebrate central nervous systems.

    PubMed Central

    Baumann, A; Grupe, A; Ackermann, A; Pongs, O

    1988-01-01

    Voltage-sensitive potassium channels are found in vertebrate and invertebrate central nervous systems. We have isolated a rat brain cDNA by cross-hybridization with a probe of the Drosophila Shaker gene complex. Structural conservation of domains of the deduced protein indicate that the rat brain cDNA encodes a voltage-sensitive potassium channel. Of the deduced amino acid sequence, 82% is homologous to the Drosophila Shaker protein indicating that voltage-sensitive potassium channels have been highly conserved during evolution. Selective pressure was highest on sequences facing the intracellular side and on proposed transmembrane segments S4-S6, suggesting that these domains are crucial for voltage-dependent potassium channel function. The corresponding rat mRNA apparently belongs to a family of mRNA molecules which are preferentially expressed in the central nervous system. Images PMID:3191911

  9. Identification and characterization of Drosophila relatives of the yeast transcriptional activator SNF2/SWI2.

    PubMed Central

    Elfring, L K; Deuring, R; McCallum, C M; Peterson, C L; Tamkun, J W

    1994-01-01

    The Drosophila brahma (brm) gene encodes an activator of homeotic genes that is highly related to the yeast transcriptional activator SWI2 (SNF2), a potential helicase. To determine whether brm is a functional homolog of SWI2 or merely a member of a family of SWI2-related genes, we searched for additional Drosophila genes related to SWI2 and examined their function in yeast cells. In addition to brm, we identified one other Drosophila relative of SWI2: the closely related ISWI gene. The 1,027-residue ISWI protein contains the DNA-dependent ATPase domain characteristic of the SWI2 protein family but lacks the three other domains common to brm and SWI2. In contrast, the ISWI protein is highly related (70% identical) to the human hSNF2L protein over its entire length, suggesting that they may be functional homologs. The DNA-dependent ATPase domains of brm and SWI2, but not ISWI, are functionally interchangeable; a chimeric SWI2-brm protein partially rescued the slow growth of swi2- cells and supported transcriptional activation mediated by the glucocorticoid receptor in vivo in yeast cells. These findings indicate that brm is the closest Drosophila relative of SWI2 and suggest that brm and SWI2 play similar roles in transcriptional activation. Images PMID:7908117

  10. The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity

    PubMed Central

    Grode, Kyle D.; Rogers, Stephen L.

    2015-01-01

    Microtubule severing is a biochemical reaction that generates an internal break in a microtubule and regulation of microtubule severing is critical for cellular processes such as ciliogenesis, morphogenesis, and meiosis and mitosis. Katanin is a conserved heterodimeric ATPase that severs and disassembles microtubules, but the molecular determinants for regulation of microtubule severing by katanin remain poorly defined. Here we show that the non-catalytic domains of Drosophila katanin regulate its abundance and activity in living cells. Our data indicate that the microtubule-interacting and trafficking (MIT) domain and adjacent linker region of the Drosophila katanin catalytic subunit Kat60 cooperate to regulate microtubule severing in two distinct ways. First, the MIT domain and linker region of Kat60 decrease its abundance by enhancing its proteasome-dependent degradation. The Drosophila katanin regulatory subunit Kat80, which is required to stabilize Kat60 in cells, conversely reduces the proteasome-dependent degradation of Kat60. Second, the MIT domain and linker region of Kat60 augment its microtubule-disassembly activity by enhancing its association with microtubules. On the basis of our data, we propose that the non-catalytic domains of Drosophila katanin serve as the principal sites of integration of regulatory inputs, thereby controlling its ability to sever and disassemble microtubules. PMID:25886649

  11. The deubiquitinating enzyme DUBAI stabilizes DIAP1 to suppress Drosophila apoptosis

    PubMed Central

    Yang, C-S; Sinenko, S A; Thomenius, M J; Robeson, A C; Freel, C D; Horn, S R; Kornbluth, S

    2014-01-01

    Deubiquitinating enzymes (DUBs) counteract ubiquitin ligases to modulate the ubiquitination and stability of target signaling molecules. In Drosophila, the ubiquitin–proteasome system has a key role in the regulation of apoptosis, most notably, by controlling the abundance of the central apoptotic regulator DIAP1. Although the mechanism underlying DIAP1 ubiquitination has been extensively studied, the precise role of DUB(s) in controlling DIAP1 activity has not been fully investigated. Here we report the identification of a DIAP1-directed DUB using two complementary approaches. First, a panel of putative Drosophila DUBs was expressed in S2 cells to determine whether DIAP1 could be stabilized, despite treatment with death-inducing stimuli that would induce DIAP1 degradation. In addition, RNAi fly lines were used to detect modifiers of DIAP1 antagonist-induced cell death in the developing eye. Together, these approaches identified a previously uncharacterized protein encoded by CG8830, which we named DeUBiquitinating-Apoptotic-Inhibitor (DUBAI), as a novel DUB capable of preserving DIAP1 to dampen Drosophila apoptosis. DUBAI interacts with DIAP1 in S2 cells, and the putative active site of its DUB domain (C367) is required to rescue DIAP1 levels following apoptotic stimuli. DUBAI, therefore, represents a novel locus of apoptotic regulation in Drosophila, antagonizing cell death signals that would otherwise result in DIAP1 degradation. PMID:24362437

  12. Analyses of interactions among pair-rule genes and the gap gene Krüppel in Bombyx segmentation.

    PubMed

    Nakao, Hajime

    2015-09-01

    In the short-germ insect Tribolium, a pair-rule gene circuit consisting of the Tribolium homologs of even-skipped, runt, and odd-skipped (Tc-eve, Tc-run and Tc-odd, respectively) has been implicated in segment formation. To examine the application of the model to other taxa, I studied the expression and function of pair-rule genes in Bombyx mori, together with a Bombyx homolog of Krüppel (Bm-Kr), a known gap gene. Knockdown embryos of Bombyx homologs of eve, run and odd (Bm-eve, Bm-run and Bm-odd) exhibited asegmental phenotypes similar to those of Tribolium knockdowns. However, pair-rule gene interactions were similar to those of both Tribolium and Drosophila, which, different from Tribolium, shows a hierarchical segmentation mode. Additionally, the Bm-odd expression pattern shares characteristics with those of Drosophila pair-rule genes that receive upstream regulatory input. On the other hand, Bm-Kr knockdowns exhibited a large posterior segment deletion as observed in short-germ insects. However, a detailed analysis of these embryos indicated that Bm-Kr modulates expression of pair-rule genes like in Drosophila, although the mechanisms appear to be different. This suggested hierarchical interactions between Bm-Kr and pair-rule genes. Based on these results, I concluded that the pair-rule gene circuit model that describes Tribolium development is not applicable to Bombyx. PMID:26102481

  13. Segmentation of Unstructured Datasets

    NASA Technical Reports Server (NTRS)

    Bhat, Smitha

    1996-01-01

    Datasets generated by computer simulations and experiments in Computational Fluid Dynamics tend to be extremely large and complex. It is difficult to visualize these datasets using standard techniques like Volume Rendering and Ray Casting. Object Segmentation provides a technique to extract and quantify regions of interest within these massive datasets. This thesis explores basic algorithms to extract coherent amorphous regions from two-dimensional and three-dimensional scalar unstructured grids. The techniques are applied to datasets from Computational Fluid Dynamics and from Finite Element Analysis.

  14. Drosophila melanogaster (fruit fly) locomotion during a sounding rocket flight

    NASA Astrophysics Data System (ADS)

    Miller, Mark S.; Keller, Tony S.

    2008-05-01

    The locomotor activity of young Drosophila melanogaster (fruit fly) was studied during a Nike-Orion sounding rocket flight, which included a short-duration microgravity exposure. An infrared monitoring system was used to determine the activity level, instantaneous velocity, and continuous velocity of 240 (120 male, 120 female) fruit flies. Individual flies were placed in chambers that limit their motion to walking. Chambers were oriented both vertically and horizontally with respect to the rocket's longitudinal axis. Significant changes in Drosophila locomotion patterns were observed throughout the sounding rocket flight, including launch, microgravity exposure, payload re-entry, and after ocean impact. During the microgravity portion of the flight (3.8 min), large increases in all locomotion measurements for both sexes were observed, with some measurements doubling compared to pad (1 G) data. Initial effects of microgravity were probably delayed due to large accelerations from the payload despining immediately before entering microgravity. The results indicate that short-duration microgravity exposure has a large effect on locomotor activity for both males and females, at least for a short period of time. The locomotion increases may explain the increased male aging observed during long-duration exposure to microgravity. Studies focusing on long-duration microgravity exposure are needed to confirm these findings, and the relationship of increased aging and locomotion.

  15. The Drosophila Helicase MLE Targets Hairpin Structures in Genomic Transcripts.

    PubMed

    Cugusi, Simona; Li, Yujing; Jin, Peng; Lucchesi, John C

    2016-01-01

    RNA hairpins are a common type of secondary structures that play a role in every aspect of RNA biochemistry including RNA editing, mRNA stability, localization and translation of transcripts, and in the activation of the RNA interference (RNAi) and microRNA (miRNA) pathways. Participation in these functions often requires restructuring the RNA molecules by the association of single-strand (ss) RNA-binding proteins or by the action of helicases. The Drosophila MLE helicase has long been identified as a member of the MSL complex responsible for dosage compensation. The complex includes one of two long non-coding RNAs and MLE was shown to remodel the roX RNA hairpin structures in order to initiate assembly of the complex. Here we report that this function of MLE may apply to the hairpins present in the primary RNA transcripts that generate the small molecules responsible for RNA interference. Using stocks from the Transgenic RNAi Project and the Vienna Drosophila Research Center, we show that MLE specifically targets hairpin RNAs at their site of transcription. The association of MLE at these sites is independent of sequence and chromosome location. We use two functional assays to test the biological relevance of this association and determine that MLE participates in the RNAi pathway. PMID:26752049

  16. Mutational Analysis of Drosophila Basigin Function in the Visual System

    PubMed Central

    Munro, Michelle; Akkam, Yazan; Curtin, Kathryn D.

    2009-01-01

    Drosophila basigin is a cell-surface glycoprotein of the Ig superfamily and a member of a protein family that includes mammalian EMMPRIN/CD147/basigin, neuroplastin, and embigin. Our previous work on Drosophila basigin has shown that it is required for normal photoreceptor cell structure and normal neuron-glia interaction in the fly visual system. Specifically, the photoreceptor neurons of mosaic animals that are mutant in the eye for basigin show altered cell structure with nuclei, mitochondria and rER misplaced and variable axon diameter compared to wild-type. In addition, glia cells in the optic lamina that contact photoreceptor axons are misplaced and show altered structure. All these defects are rescued by expression of either transgenic fly basigin or transgenic mouse basigin in the photoreceptors demonstrating that mouse basigin can functionally replace fly basigin. To determine what regions of the basigin protein are required for each of these functions, we have created mutant basigin transgenes coding for proteins that are altered in conserved residues, introduced these into the fly genome, and tested them for their ability to rescue both photoreceptor cell structure defects and neuron-glia interaction defects of basigin. The results suggest that the highly conserved transmembrane domain and the extracellular domains are crucial for basigin function in the visual system while the short intracellular tail may not play a role in these functions. PMID:19782733

  17. Mutational analysis of Drosophila basigin function in the visual system.

    PubMed

    Munro, Michelle; Akkam, Yazan; Curtin, Kathryn D

    2010-01-01

    Drosophila basigin is a cell-surface glycoprotein of the Ig superfamily and a member of a protein family that includes mammalian EMMPRIN/CD147/basigin, neuroplastin, and embigin. Our previous work on Drosophila basigin has shown that it is required for normal photoreceptor cell structure and normal neuron-glia interaction in the fly visual system. Specifically, the photoreceptor neurons of mosaic animals that are mutant in the eye for basigin show altered cell structure with nuclei, mitochondria and rER misplaced and variable axon diameter compared to wild-type. In addition, glia cells in the optic lamina that contact photoreceptor axons are misplaced and show altered structure. All these defects are rescued by expression of either transgenic fly basigin or transgenic mouse basigin in the photoreceptors demonstrating that mouse basigin can functionally replace fly basigin. To determine what regions of the basigin protein are required for each of these functions, we have created mutant basigin transgenes coding for proteins that are altered in conserved residues, introduced these into the fly genome, and tested them for their ability to rescue both photoreceptor cell structure defects and neuron-glia interaction defects of basigin. The results suggest that the highly conserved transmembrane domain and the extracellular domains are crucial for basigin function in the visual system while the short intracellular tail may not play a role in these functions. PMID:19782733

  18. Sex-specific regulation of Lgr3 in Drosophila neurons

    PubMed Central

    Meissner, Geoffrey W.; Luo, Shengzhan D.; Dias, Brian G.; Texada, Michael J.; Baker, Bruce S.

    2016-01-01

    The development of sexually dimorphic morphology and the potential for sexually dimorphic behavior in Drosophila are regulated by the Fruitless (Fru) and Doublesex (Dsx) transcription factors. Several direct targets of Dsx have been identified, but direct Fru targets have not been definitively identified. We show that Drosophila leucine-rich repeat G protein-coupled receptor 3 (Lgr3) is regulated by Fru and Dsx in separate populations of neurons. Lgr3 is a member of the relaxin-receptor family and a receptor for Dilp8, necessary for control of organ growth. Lgr3 expression in the anterior central brain of males is inhibited by the B isoform of Fru, whose DNA binding domain interacts with a short region of an Lgr3 intron. Fru A and C isoform mutants had no observed effect on Lgr3 expression. The female form of Dsx (DsxF) separately up- and down-regulates Lgr3 expression in distinct neurons in the abdominal ganglion through female- and male-specific Lgr3 enhancers. Excitation of neural activity in the DsxF–up-regulated abdominal ganglion neurons inhibits female receptivity, indicating the importance of these neurons for sexual behavior. Coordinated regulation of Lgr3 by Fru and Dsx marks a point of convergence of the two branches of the sex-determination hierarchy. PMID:26884206

  19. Glucose modulates Drosophila longevity and immunity independent of the microbiota

    PubMed Central

    Galenza, Anthony; Hutchinson, Jaclyn; Campbell, Shelagh D.; Hazes, Bart; Foley, Edan

    2016-01-01

    ABSTRACT The acquisition of nutrients is essential for maintenance of metabolic processes in all organisms. Nutritional imbalance contributes to myriad metabolic disorders that include malnutrition, diabetes and even cancer. Recently, the importance of macronutrient ratio of food has emerged as a critical factor to determine health outcomes. Here we show that individual modifications to a completely defined diet markedly impact multiple aspects of organism wellbeing in Drosophila melanogaster. Through a longitudinal survey of several diets we demonstrate that increased levels of dietary glucose significantly improve longevity and immunity in adult Drosophila. Our metagenomic studies show that relative macronutrient levels not only influence the host, but also have a profound impact on microbiota composition. However, we found that elevated dietary glucose extended the lifespan of adult flies even when raised in a germ-free environment. Furthermore, when challenged with a chronic enteric infection, flies fed a diet with added glucose had increased survival times even in the absence of an intact microbiota. Thus, in contrast to known links between the microbiota and animal health, our findings uncover a novel microbiota-independent response to diet that impacts host wellbeing. As dietary responses are highly conserved in animals, we believe our results offer a general understanding of the association between glucose metabolism and animal health. PMID:26794610

  20. The Drosophila Helicase MLE Targets Hairpin Structures in Genomic Transcripts

    PubMed Central

    Cugusi, Simona; Li, Yujing; Jin, Peng; Lucchesi, John C.

    2016-01-01

    RNA hairpins are a common type of secondary structures that play a role in every aspect of RNA biochemistry including RNA editing, mRNA stability, localization and translation of transcripts, and in the activation of the RNA interference (RNAi) and microRNA (miRNA) pathways. Participation in these functions often requires restructuring the RNA molecules by the association of single-strand (ss) RNA-binding proteins or by the action of helicases. The Drosophila MLE helicase has long been identified as a member of the MSL complex responsible for dosage compensation. The complex includes one of two long non-coding RNAs and MLE was shown to remodel the roX RNA hairpin structures in order to initiate assembly of the complex. Here we report that this function of MLE may apply to the hairpins present in the primary RNA transcripts that generate the small molecules responsible for RNA interference. Using stocks from the Transgenic RNAi Project and the Vienna Drosophila Research Center, we show that MLE specifically targets hairpin RNAs at their site of transcription. The association of MLE at these sites is independent of sequence and chromosome location. We use two functional assays to test the biological relevance of this association and determine that MLE participates in the RNAi pathway. PMID:26752049