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Sample records for drosophila larval development

  1. The development of the Drosophila larval brain.

    PubMed

    Hartenstein, Volker; Spindler, Shana; Pereanu, Wayne; Fung, Siaumin

    2008-01-01

    In this chapter we will start out by describing in more detail the progenitors of the nervous system, the neuroblasts and ganglion mother cells. Subsequently we will survey the generic cell types that make up the developing Drosophila brain, namely neurons, glial cells and tracheal cells. Finally, we will attempt a synopsis of the neuronal connectivity of the larval brain that can be deduced from the analysis of neural lineages and their relationship to neuropile compartments. PMID:18683635

  2. Development and plasticity of the Drosophila larval neuromuscular junction

    PubMed Central

    Menon, Kaushiki P.; Carrillo, Robert A.; Zinn, Kai

    2013-01-01

    The Drosophila larval neuromuscular system is relatively simple, containing only 32 motor neurons in each abdominal hemisegment, and its neuromuscular junctions (NMJs) are large, individually specified, and easy to visualize and record from. NMJ synapses exhibit developmental and functional plasticity while displaying stereotyped connectivity. Drosophila Type I NMJ synapses are glutamatergic, while the vertebrate NMJ uses acetylcholine as its primary neurotransmitter. The larval NMJ synapses use ionotropic glutamate receptors (GluRs) that are homologous to AMPA-type glutamate receptors in the mammalian brain, and they have postsynaptic scaffolds that resemble those found in mammalian postsynaptic densities. These features make the Drosophila neuromuscular system an excellent genetic model for the study of excitatory synapses in the mammalian central nervous system. The first section of the review presents an overview of NMJ development. The second section describes genes that regulate NMJ development, including: 1) genes that positively and negatively regulate growth of the NMJ; 2) genes required for maintenance of NMJ bouton structure; 3) genes that modulate neuronal activity and alter NMJ growth; 4) genes involved in trans-synaptic signaling at the NMJ. The third section describes genes that regulate acute plasticity, focusing on translational regulatory mechanisms. Since this review is intended for a developmental biology audience, it does not cover NMJ electrophysiology in detail, and does not review genes for which mutations produce only electrophysiological but no structural phenotypes. PMID:24014452

  3. 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

  4. Muscle organizers in Drosophila: the role of persistent larval fibers in adult flight muscle development

    NASA Technical Reports Server (NTRS)

    Farrell, E. R.; Fernandes, J.; Keshishian, H.

    1996-01-01

    In many organisms muscle formation depends on specialized cells that prefigure the pattern of the musculature and serve as templates for myoblast organization and fusion. These include muscle pioneers in insects and muscle organizing cells in leech. In Drosophila, muscle founder cells have been proposed to play a similar role in organizing larval muscle development during embryogenesis. During metamorphosis in Drosophila, following histolysis of most of the larval musculature, there is a second round of myogenesis that gives rise to the adult muscles. It is not known whether muscle founder cells organize the development of these muscles. However, in the thorax specific larval muscle fibers do not histolyze at the onset of metamorphosis, but instead serve as templates for the formation of a subset of adult muscles, the dorsal longitudinal flight muscles (DLMs). Because these persistent larval muscle fibers appear to be functioning in many respects like muscle founder cells, we investigated whether they were necessary for DLM development by using a microbeam laser to ablate them singly and in combination. We found that, in the absence of the larval muscle fibers, DLMs nonetheless develop. Our results show that the persistent larval muscle fibers are not required to initiate myoblast fusion, to determine DLM identity, to locate the DLMs in the thorax, or to specify the total DLM fiber volume. However, they are required to regulate the number of DLM fibers generated. Thus, while the persistent larval muscle fibers are not obligatory for DLM fiber formation and differentiation, they are necessary to ensure the development of the correct number of fibers.

  5. Persistent Activation of the Innate Immune Response in Adult Drosophila Following Radiation Exposure During Larval Development

    PubMed Central

    Sudmeier, Lisa J.; Samudrala, Sai-Suma; Howard, Steven P.; Ganetzky, Barry

    2015-01-01

    Cranial radiation therapy (CRT) is an effective treatment for pediatric central nervous system malignancies, but survivors often suffer from neurological and neurocognitive side effects that occur many years after radiation exposure. Although the biological mechanisms underlying these deleterious side effects are incompletely understood, radiation exposure triggers an acute inflammatory response that may evolve into chronic inflammation, offering one avenue of investigation. Recently, we developed a Drosophila model of the neurotoxic side effects of radiation exposure. Here we use this model to investigate the role of the innate immune system in response to radiation exposure. We show that the innate immune response and NF-ĸB target gene expression is activated in the adult Drosophila brain following radiation exposure during larval development, and that this response is sustained in adult flies weeks after radiation exposure. We also present preliminary data suggesting that innate immunity is radioprotective during Drosophila development. Together our data suggest that activation of the innate immune response may be beneficial initially for survival following radiation exposure but result in long-term deleterious consequences, with chronic inflammation leading to impaired neuronal function and viability at later stages. This work lays the foundation for future studies of how the innate immune response is triggered by radiation exposure and its role in mediating the biological responses to radiation. These studies may facilitate the development of strategies to reduce the deleterious side effects of CRT. PMID:26333838

  6. amontillado, the Drosophila homolog of the prohormone processing protease PC2, is required during embryogenesis and early larval development.

    PubMed Central

    Rayburn, Lowell Y M; Gooding, Holly C; Choksi, Semil P; Maloney, Dhea; Kidd, Ambrose R; Siekhaus, Daria E; Bender, Michael

    2003-01-01

    Biosynthesis of most peptide hormones and neuropeptides requires proteolytic excision of the active peptide from inactive proprotein precursors, an activity carried out by subtilisin-like proprotein convertases (SPCs) in constitutive or regulated secretory pathways. The Drosophila amontillado (amon) gene encodes a homolog of the mammalian PC2 protein, an SPC that functions in the regulated secretory pathway in neuroendocrine tissues. We have identified amon mutants by isolating ethylmethanesulfonate (EMS)-induced lethal and visible mutations that define two complementation groups in the amon interval at 97D1 of the third chromosome. DNA sequencing identified the amon complementation group and the DNA sequence change for each of the nine amon alleles isolated. amon mutants display partial embryonic lethality, are defective in larval growth, and arrest during the first to second instar larval molt. Mutant larvae can be rescued by heat-shock-induced expression of the amon protein. Rescued larvae arrest at the subsequent larval molt, suggesting that amon is also required for the second to third instar larval molt. Our data indicate that the amon proprotein convertase is required during embryogenesis and larval development in Drosophila and support the hypothesis that AMON acts to proteolytically process peptide hormones that regulate hatching, larval growth, and larval ecdysis. PMID:12586710

  7. '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

  8. Drosophila adult and larval pheromones modulate larval food choice

    PubMed Central

    Farine, Jean-Pierre; Cortot, Jérôme; Ferveur, Jean-François

    2014-01-01

    Insects use chemosensory cues to feed and mate. In Drosophila, the effect of pheromones has been extensively investigated in adults, but rarely in larvae. The colonization of natural food sources by Drosophila buzzatii and Drosophila simulans species may depend on species-specific chemical cues left in the food by larvae and adults. We identified such chemicals in both species and measured their influence on larval food preference and puparation behaviour. We also tested compounds that varied between these species: (i) two larval volatile compounds: hydroxy-3-butanone-2 and phenol (predominant in D. simulans and D. buzzatii, respectively), and (ii) adult cuticular hydrocarbons (CHs). Drosophila buzzatii larvae were rapidly attracted to non-CH adult conspecific cues, whereas D. simulans larvae were strongly repulsed by CHs of the two species and also by phenol. Larval cues from both species generally reduced larval attraction and pupariation on food, which was generally—but not always—low, and rarely reflected larval response. As these larval and adult pheromones specifically influence larval food search and the choice of a pupariation site, they may greatly affect the dispersion and survival of Drosophila species in nature. PMID:24741012

  9. Sternopleural is a regulatory mutation of wingless with both dominant and recessive effects on larval development of Drosophila melanogaster

    SciTech Connect

    Neumann, C.J.; Cohen, S.M.

    1996-04-01

    The Drosophila wingless (wg) gene encodes a secreted signaling protein that is required for many separate patterning events in both embryonic and larval development. wg functions in the development of the adult structures have been studied using the conditional mutant wg{sup ts} and also using regulatory mutations of wg that reduce larval functions. Here we present evidence that Sternopleural (Sp) is another regulatory allele of wg that affects a subset of larval functions. Sp has both a recessive loss-of-function component and a gain-of-function component. The loss-of-function component reflects a reduction of wg activity in the notum and in the antenna. The gain-of-function component apparently leads to ectopic wg activity in the dorsal first and second leg disc and thereby generates the dominant Sp phenotype. Sp and other wg alleles show a complex pattern of complementation. We present evidence that these genetic properties are due to transvection. These results have implications for the genetic definition of a null allele at loci subject to transvection. 31 refs., 6 figs., 1 tab.

  10. Effects of Three Volatile Oxylipins on Colony Development in Two Species of Fungi and on Drosophila Larval Metamorphosis.

    PubMed

    Yin, Guohua; Padhi, Sally; Lee, Samantha; Hung, Richard; Zhao, Guozhu; Bennett, Joan W

    2015-09-01

    The aim of this study is to investigate the effects of three volatile oxylipins on colony development in two fungi and on Drosophila larval metamorphosis. Using an airborne exposure technique, three common and volatile oxylipins (1-octen-3-ol, (E)-2-hexenal, and 1-hexanol) were compared for their effects on spore germination and colony growth in Aspergillus niger and Penicillium chrysogenum, as well as for their effects on the morphogenesis of larvae of Drosophila melanogaster. Conidia of both A. niger and P. chrysogenum plated in the presence of low concentrations (50 ppm) of these three volatile organic compounds (VOCs) formed fewer colony-forming units (CFUs) and exhibited reduced radial growth of colonies as compared to controls. When A. niger and P. chrysogenum spores were germinated in the presence of the enantiomers of 1-octen-3-ol, (R)-(-)-1-octen-3-ol had the greatest impact on colony morphology (decreased sporulation and colony diameter), while (S)-(+)-1-octen-3-ol and the racemic form yielded similar morphological changes but to a lesser extent. In addition, Drosophila larvae exposed to vapors of these oxylipins exhibited serious delays in metamorphosis and toxic effects on pupae and adult stages. Low concentration of these three VOCs can significantly inhibit the formation of CFUs and the growth of fungi. (R)-(-)-1-octen-3-ol imposed the greatest impact on fungal morphology compared to (S)-(+)-1-octen-3-ol and the racemic form. The three volatile oxylipins could also delay the metamorphosis of Drosophila and impose toxic effects on its pupae and adult stages. PMID:26126831

  11. Patterns of growth and tract formation during the early development of secondary lineages in the Drosophila larval brain.

    PubMed

    Lovick, Jennifer K; Kong, Angel; Omoto, Jaison J; Ngo, Kathy T; Younossi-Hartenstein, Amelia; Hartenstein, Volker

    2016-04-01

    The Drosophila brain consists of a relatively small number of invariant, genetically determined lineages which provide a model to study the relationship between gene function and neuronal architecture. In following this long-term goal, we reconstruct the morphology (projection pattern and connectivity) and gene expression patterns of brain lineages throughout development. In this article, we focus on the secondary phase of lineage morphogenesis, from the reactivation of neuroblast proliferation in the first larval instar to the time when proliferation ends and secondary axon tracts have fully extended in the late third larval instar. We have reconstructed the location and projection of secondary lineages at close (4 h) intervals and produced a detailed map in the form of confocal z-projections and digital three-dimensional models of all lineages at successive larval stages. Based on these reconstructions, we could compare the spatio-temporal pattern of axon formation and morphogenetic movements of different lineages in normal brain development. In addition to wild type, we reconstructed lineage morphology in two mutant conditions. (1) Expressing the construct UAS-p35 which rescues programmed cell death we could systematically determine which lineages normally lose hemilineages to apoptosis. (2) so-Gal4-driven expression of dominant-negative EGFR ablated the optic lobe, which allowed us to conclude that the global centrifugal movement normally affecting the cell bodies of lateral lineages in the late larva is causally related to the expansion of the optic lobe, and that the central pattern of axonal projections of these lineages is independent of the presence or absence of the optic lobe. PMID:26178322

  12. Drosophila cortex and neuropile glia influence secondary axon tract growth, pathfinding, and fasciculation in the developing larval brain.

    PubMed

    Spindler, Shana R; Ortiz, Irma; Fung, Siaumin; Takashima, Shigeo; Hartenstein, Volker

    2009-10-15

    Glial cells play important roles in the developing brain during axon fasciculation, growth cone guidance, and neuron survival. In the Drosophila brain, three main classes of glia have been identified including surface, cortex, and neuropile glia. While surface glia ensheaths the brain and is involved in the formation of the blood-brain-barrier and the control of neuroblast proliferation, the range of functions for cortex and neuropile glia is less well understood. In this study, we use the nirvana2-GAL4 driver to visualize the association of cortex and neuropile glia with axon tracts formed by different brain lineages and selectively eliminate these glial populations via induced apoptosis. The larval central brain consists of approximately 100 lineages. Each lineage forms a cohesive axon bundle, the secondary axon tract (SAT). While entering and traversing the brain neuropile, SATs interact in a characteristic way with glial cells. Some SATs are completely invested with glial processes; others show no particular association with glia, and most fall somewhere in between these extremes. Our results demonstrate that the elimination of glia results in abnormalities in SAT fasciculation and trajectory. The most prevalent phenotype is truncation or misguidance of axon tracts, or abnormal fasciculation of tracts that normally form separate pathways. Importantly, the degree of glial association with a given lineage is positively correlated with the severity of the phenotype resulting from glial ablation. Previous studies have focused on the embryonic nerve cord or adult-specific compartments to establish the role of glia. Our study provides, for the first time, an analysis of glial function in the brain during axon formation and growth in larval development. PMID:19646433

  13. Drosophila cortex and neuropile glia influence secondary axon tract growth, pathfinding, and fasciculation in the developing larval brain

    PubMed Central

    Spindler, Shana R; Ortiz, Irma; Fung, Siaumin; Takashima, Shigeo; Hartenstein, Volker

    2009-01-01

    Glial cells play important roles in the developing brain during axon fasciculation, growth cone guidance, and neuron survival. In the Drosophila brain, three main classes of glia have been identified including surface, cortex, and neuropile glia. While surface glia ensheaths the brain and is involved in the formation of the blood-brain-barrier and the control of neuroblast proliferation, the range of functions for cortex and neuropile glia is less well understood. In this study, we use the nirvana2-GAL4 driver to visualize the association of cortex and neuropile glia with axon tracts formed by different brain lineages and selectively eliminate these glial populations via induced apoptosis. The larval central brain consists of approximately 100 lineages. Each lineage forms a cohesive axon bundle, the secondary axon tract (SAT). While entering and traversing the brain neuropile, SATs interact in a characteristic way with glial cells. Some SATs are completely invested with glial processes; others show no particular association with glia, and most fall somewhere in between these extremes. Our results demonstrate that the elimination of glia results in abnormalities in SAT fasciculation and trajectory. The most prevalent phenotype is truncation or misguidance of axon tracts, or abnormal fasciculation of tracts that normally form separate pathways. Importantly, the degree of glial association with a given lineage is positively correlated with the severity of the phenotype resulting from glial ablation. Previous studies have focused on the embryonic nerve cord or adult specific compartments to establish the role of glia. Our study provides, for the first time, an analysis of glial function in the brain during axon formation and growth in larval development. PMID:19646433

  14. ‘Peer pressure’ in larval Drosophila?

    PubMed Central

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

    2014-01-01

    ABSTRACT 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

  15. Transgenerational Effects of Parental Larval Diet on Offspring Development Time, Adult Body Size and Pathogen Resistance in Drosophila melanogaster

    PubMed Central

    Valtonen, Terhi M.; Kangassalo, Katariina; Pölkki, Mari; Rantala, Markus J.

    2012-01-01

    Environmental conditions experienced by parents are increasingly recognized to affect offspring performance. We set out to investigate the effect of parental larval diet on offspring development time, adult body size and adult resistance to the bacterium Serratia marcescens in Drosophila melanogaster. Flies for the parental generation were raised on either poor or standard diet and then mated in the four possible sex-by-parental diet crosses. Females that were raised on poor food produced larger offspring than females that were raised on standard food. Furthermore, male progeny sired by fathers that were raised on poor food were larger than male progeny sired by males raised on standard food. Development times were shortest for offspring whose one parent (mother or the father) was raised on standard and the other parent on poor food and longest for offspring whose parents both were raised on poor food. No evidence for transgenerational effects of parental diet on offspring disease resistance was found. Although paternal effects have been previously demonstrated in D. melanogaster, no earlier studies have investigated male-mediated transgenerational effects of diet in this species. The results highlight the importance of not only considering the relative contribution each parental sex has on progeny performance but also the combined effects that the two sexes may have on offspring performance. PMID:22359607

  16. Gonadosomatic mosaicism for lethal mutations in Drosophila lethal mutations disturbing larval development

    SciTech Connect

    Ivanov, A.I.; Sakharova, N.Yu.

    1988-11-01

    Phenogenetic analysis of autonomous lethal mutations obtained by the method of gonadosomatic mosaicism which manifested during larval stages, established that the nuclei of hypodermal cells, salivary glands suprapharyngeal ganglion, pharynx, esophagus, gizzard, and hindgut are the derivatives of the same nucleus (from the first two nuclei of cleavage) as the nuclei of the cells of the imaginal-somatic tissues.

  17. Origin and development of neuropil glia of the Drosophila larval and adult brain: two distinct glial populations derived from separate progenitors

    PubMed Central

    Omoto, Jaison Jiro; Yogi, Puja; Hartenstein, Volker

    2015-01-01

    Glia comprise a conspicuous population of non-neuronal cells in vertebrate and invertebrate nervous systems. Drosophila serves as a favorable model to elucidate basic principles of glial biology in vivo. The Drosophila neuropil glia (NPG), subdivided into astrocyte-like (ALG) and ensheathing glia (EG), extend reticular processes which associate with synapses and sheath-like processes which surround neuropil compartments, respectively. In this paper we characterize the development of NPG throughout fly brain development. We find that differentiated neuropil glia of the larval brain originate as a cluster of precursors derived from embryonic progenitors located in the basal brain. These precursors undergo a characteristic migration to spread over the neuropil surface while specifying/differentiating into primary ALG and EG. Embryonically-derived primary NPG are large cells which are few in number, and occupy relatively stereotyped positions around the larval neuropil surface. During metamorphosis, primary NPG undergo cell death. Neuropil glia of the adult (secondary NPG) are derived from type II lineages during the postembryonic phase of neurogliogenesis. These secondary NPG are much smaller in size but greater in number than primary NPG. Lineage tracing reveals that both NPG subtypes derive from intermediate neural progenitors of multipotent type II lineages. Taken together, this study reveals previously uncharacterized dynamics of NPG development and provides a framework for future studies utilizing Drosophila glia as a model. PMID:25779704

  18. Origin and development of neuropil glia of the Drosophila larval and adult brain: Two distinct glial populations derived from separate progenitors.

    PubMed

    Omoto, Jaison Jiro; Yogi, Puja; Hartenstein, Volker

    2015-08-15

    Glia comprise a conspicuous population of non-neuronal cells in vertebrate and invertebrate nervous systems. Drosophila serves as a favorable model to elucidate basic principles of glial biology in vivo. The Drosophila neuropil glia (NPG), subdivided into astrocyte-like (ALG) and ensheathing glia (EG), extend reticular processes which associate with synapses and sheath-like processes which surround neuropil compartments, respectively. In this paper we characterize the development of NPG throughout fly brain development. We find that differentiated neuropil glia of the larval brain originate as a cluster of precursors derived from embryonic progenitors located in the basal brain. These precursors undergo a characteristic migration to spread over the neuropil surface while specifying/differentiating into primary ALG and EG. Embryonically-derived primary NPG are large cells which are few in number, and occupy relatively stereotyped positions around the larval neuropil surface. During metamorphosis, primary NPG undergo cell death. Neuropil glia of the adult (secondary NPG) are derived from type II lineages during the postembryonic phase of neurogliogenesis. These secondary NPG are much smaller in size but greater in number than primary NPG. Lineage tracing reveals that both NPG subtypes derive from intermediate neural progenitors of multipotent type II lineages. Taken together, this study reveals previously uncharacterized dynamics of NPG development and provides a framework for future studies utilizing Drosophila glia as a model. PMID:25779704

  19. Contribution of larval nutrition to adult reproduction in Drosophila melanogaster.

    PubMed

    Aguila, Jerell R; Hoshizaki, Deborah K; Gibbs, Allen G

    2013-02-01

    Within the complex life cycle of holometabolous insects, nutritional resources acquired during larval feeding are utilized by the pupa and the adult. The broad features of the transfer of larval resources to the pupae and the allocation of larval resources in the adult have been described by studies measuring and tracking macronutrients at different developmental stages. However, the mechanisms of resource transfer from the larva and the factors regulating the allocation of these resources in the adult between growth, reproduction and somatic maintenance are unknown. Drosophila melanogaster presents a tractable system in which to test cellular and tissue mechanisms of resource acquisition and allocation because of the detailed understanding of D. melanogaster development and the experimental tools to manipulate its tissues across developmental stages. In previous work, we demonstrated that the fat body of D. melanogaster larvae is important for survival of starvation stress in the young adult, and suggested that programmed cell death of the larval fat cells in the adult is important for allocation of resources for female reproduction. Here, we describe the temporal uptake of larval-derived carbon by the ovaries, and demonstrate the importance of larval fat-cell death in the maturation of the ovary and in fecundity. Larvae and adults were fed stable carbon isotopes to follow the acquisition of larval-derived carbon by the adult ovaries. We determined that over half of the nutrients acquired by the ovaries in 2-day-old adult females are dependent upon the death of the fat cells. Furthermore, when programmed cell death is inhibited in the larval fat cells, ovarian development was depressed and fecundity was reduced. PMID:23038728

  20. 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

  1. 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

  2. Dissection of larval CNS in Drosophila melanogaster.

    PubMed

    Hafer, Nathaniel; Schedl, Paul

    2006-12-01

    The central nervous system (CNS) of Drosophila larvae is complex and poorly understood. One way to investigate the CNS is to use immunohistochemistry to examine the expression of various novel and marker proteins. Staining of whole larvae is impractical because the tough cuticle prevents antibodies from penetrating inside the body cavity. In order to stain these tissues it is necessary to dissect the animal prior to fixing and staining. In this article we demonstrate how to dissect Drosophila larvae without damaging the CNS. Begin by tearing the larva in half with a pair of fine forceps, and then turn the cuticle "inside-out" to expose the CNS. If the dissection is performed carefully the CNS will remain attached to the cuticle. We usually keep the CNS attached to the cuticle throughout the fixation and staining steps, and only completely remove the CNS from the cuticle just prior to mounting the samples on glass slides. We also show some representative images of a larval CNS stained with Eve, a transcription factor expressed in a subset of neurons in the CNS. The article concludes with a discussion of some of the practical uses of this technique and the potential difficulties that may arise. PMID:18704179

  3. FACS purification of Drosophila larval Neuroblasts for next generation sequencing

    PubMed Central

    Conder, Ryan; Schmauss, Gerald; Knoblich, Juergen A.

    2014-01-01

    Elegant tools are available for the genetic analysis of neural stem cell lineages in Drosophila, but a methodology for purifying stem cells and their differentiated progeny for transcriptome analysis is currently missing. Previous attempts to overcome this problem either involved using RNA isolated from whole larval brain tissue or co-transcriptional in vivo mRNA tagging. As both methods have limited cell type specificity, we developed a protocol for the isolation of Drosophila neural stem cells (neuroblasts, NBs) and their differentiated sibling cells by FACS. We dissected larval brains from fly strains expressing GFP under the control of a NB lineage-specific GAL4 line. Upon dissociation, we made use of differences in GFP intensity and cell size to separate NBs and neurons. The resulting cell populations are over 98% pure and can readily be used for live imaging or gene expression analysis. Our method is optimized for neural stem cells, but it can also be applied to other Drosophila cell types. Primary cell suspensions and sorted cell populations can be obtained within 1 d; material for deep-sequencing library preparation can be obtained within 4 d. PMID:23660757

  4. Transmembrane channel-like (tmc) gene regulates Drosophila larval locomotion.

    PubMed

    Guo, Yanmeng; Wang, Yuping; Zhang, Wei; Meltzer, Shan; Zanini, Damiano; Yu, Yue; Li, Jiefu; Cheng, Tong; Guo, Zhenhao; Wang, Qingxiu; Jacobs, Julie S; Sharma, Yashoda; Eberl, Daniel F; Göpfert, Martin C; Jan, Lily Yeh; Jan, Yuh Nung; Wang, Zuoren

    2016-06-28

    Drosophila larval locomotion, which entails rhythmic body contractions, is controlled by sensory feedback from proprioceptors. The molecular mechanisms mediating this feedback are little understood. By using genetic knock-in and immunostaining, we found that the Drosophila melanogaster transmembrane channel-like (tmc) gene is expressed in the larval class I and class II dendritic arborization (da) neurons and bipolar dendrite (bd) neurons, both of which are known to provide sensory feedback for larval locomotion. Larvae with knockdown or loss of tmc function displayed reduced crawling speeds, increased head cast frequencies, and enhanced backward locomotion. Expressing Drosophila TMC or mammalian TMC1 and/or TMC2 in the tmc-positive neurons rescued these mutant phenotypes. Bending of the larval body activated the tmc-positive neurons, and in tmc mutants this bending response was impaired. This implicates TMC's roles in Drosophila proprioception and the sensory control of larval locomotion. It also provides evidence for a functional conservation between Drosophila and mammalian TMCs. PMID:27298354

  5. Adaptation to larval crowding in Drosophila ananassae and Drosophila nasuta nasuta: increased larval competitive ability without increased larval feeding rate.

    PubMed

    Nagarajan, Archana; Natarajan, Sharmila Bharathi; Jayaram, Mohan; Thammanna, Ananda; Chari, Sudarshan; Bose, Joy; Jois, Shreyas V; Joshi, Amitabh

    2016-06-01

    The standard view of adaptation to larval crowding in fruitflies, built on results from 25 years of multiple experimental evolution studies on Drosophila melanogaster, was that enhanced competitive ability evolves primarily through increased larval feeding and foraging rate, and increased larval tolerance to nitrogenous wastes, at the cost of efficiency of food conversion to biomass. These results were at odds from the predictions of classical K-selection theory, notably the expectation that selection at high density should result in the increase of efficiency of conversion of food to biomass, and were better interpreted through the lens of α-selection. We show here that populations of D. ananassae and D. n. nasuta subjected to extreme larval crowding evolve greater competitive ability and pre-adult survivorship at high density, primarily through a combination of reduced larval duration, faster attainment of minimum critical size for pupation, greater time efficiency of food conversion to biomass and increased pupation height, with a relatively small role of increased urea/ammonia tolerance, if at all. This is a very different suite of traits than that seen to evolve under similar selection in D. melanogaster, and seems to be closer to the expectations from the canonical theory of K-selection. We also discuss possible reasons for these differences in results across the three species. Overall, the results reinforce the view that our understanding of the evolution of competitive ability in fruitflies needs to be more nuanced than before, with an appreciation that there may be multiple evolutionary routes through which higher competitive ability can be attained. PMID:27350686

  6. Measurement of Larval Activity in the Drosophila Activity Monitor

    PubMed Central

    McParland, Aidan L.; Follansbee, Taylor L.; Ganter, Geoffrey K.

    2016-01-01

    Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This protocol repurposes an instrument often used to measure adult activity, the TriKinetics Drosophila activity monitor (MB5 Multi-Beam Activity Monitor) to study larval activity. The instrument can monitor the movements of animals in 16 individual 8 cm glass assay tubes, using 17 infrared detection beams per tube. Logging software automatically saves data to a computer, recording parameters such as number of moves, times sensors were triggered, and animals’ positions within the tubes. The data can then be analyzed to represent overall locomotion and/or position preference as well as other measurements. All data are easily accessible and compatible with basic graphing and data manipulation software. This protocol will discuss how to use the apparatus, how to operate the software and how to run a larval activity assay from start to finish. PMID:25993121

  7. The Drosophila melanogaster importin alpha3 locus encodes an essential gene required for the development of both larval and adult tissues.

    PubMed Central

    Mason, D Adam; Máthé, Endre; Fleming, Robert J; Goldfarb, David S

    2003-01-01

    The nuclear transport of classical nuclear localization signal (cNLS)-containing proteins is mediated by the cNLS receptor importin alpha. The conventional importin alpha gene family in metazoan animals is composed of three clades that are conserved between flies and mammals and are referred to here as alpha1, alpha2, and alpha3. In contrast, plants and fungi contain only alpha1 genes. In this study we report that Drosophila importin alpha3 is required for the development of both larval and adult tissues. Importin alpha3 mutant flies die around the transition from first to second instar larvae, and homozygous importin alpha3 mutant eyes are defective. The transition to second instar larvae was rescued with importin alpha1, alpha2, or alpha3 transgenes, indicating that Importin alpha3 is normally required at this stage for an activity shared by all three importin alpha's. In contrast, an alpha3-specific biochemical activity(s) of Importin alpha3 is probably required for development to adults and photoreceptor cell development, since only an importin alpha3 transgene rescued these processes. These results are consistent with the view that the importin alpha's have both overlapping and distinct functions and that their role in animal development involves the spatial and temporal control of their expression. PMID:14704178

  8. Learning the specific quality of taste reinforcement in larval Drosophila

    PubMed Central

    Schleyer, Michael; Miura, Daisuke; Tanimura, Teiichi; Gerber, Bertram

    2015-01-01

    The only property of reinforcement insects are commonly thought to learn about is its value. We show that larval Drosophila not only remember the value of reinforcement (How much?), but also its quality (What?). This is demonstrated both within the appetitive domain by using sugar vs amino acid as different reward qualities, and within the aversive domain by using bitter vs high-concentration salt as different qualities of punishment. From the available literature, such nuanced memories for the quality of reinforcement are unexpected and pose a challenge to present models of how insect memory is organized. Given that animals as simple as larval Drosophila, endowed with but 10,000 neurons, operate with both reinforcement value and quality, we suggest that both are fundamental aspects of mnemonic processing—in any brain. DOI: http://dx.doi.org/10.7554/eLife.04711.001 PMID:25622533

  9. Patterning the dorsal longitudinal flight muscles (DLM) of Drosophila: insights from the ablation of larval scaffolds

    NASA Technical Reports Server (NTRS)

    Fernandes, J. J.; Keshishian, H.

    1996-01-01

    The six Dorsal Longitudinal flight Muscles (DLMs) of Drosophila develop from three larval muscles that persist into metamorphosis and serve as scaffolds for the formation of the adult fibers. We have examined the effect of muscle scaffold ablation on the development of DLMs during metamorphosis. Using markers that are specific to muscle and myoblasts we show that in response to the ablation, myoblasts which would normally fuse with the larval muscle, fuse with each other instead, to generate the adult fibers in the appropriate regions of the thorax. The development of these de novo DLMs is delayed and is reflected in the delayed expression of erect wing, a transcription factor thought to control differentiation events associated with myoblast fusion. The newly arising muscles express the appropriate adult-specific Actin isoform (88F), indicating that they have the correct muscle identity. However, there are frequent errors in the number of muscle fibers generated. Ablation of the larval scaffolds for the DLMs has revealed an underlying potential of the DLM myoblasts to initiate de novo myogenesis in a manner that resembles the mode of formation of the Dorso-Ventral Muscles, DVMs, which are the other group of indirect flight muscles. Therefore, it appears that the use of larval scaffolds is a superimposition on a commonly used mechanism of myogenesis in Drosophila. Our results show that the role of the persistent larval muscles in muscle patterning involves the partitioning of DLM myoblasts, and in doing so, they regulate formation of the correct number of DLM fibers.

  10. Genetic and evolutionary analysis of the Drosophila larval neuromuscular junction

    NASA Astrophysics Data System (ADS)

    Campbell, Megan

    Although evolution of brains and behaviors is of fundamental biological importance, we lack comprehensive understanding of the general principles governing these processes or the specific mechanisms and molecules through which the evolutionary changes are effected. Because synapses are the basic structural and functional units of nervous systems, one way to address these problems is to dissect the genetic and molecular pathways responsible for morphological evolution of a defined synapse. I have undertaken such an analysis by examining morphology of the larval neuromuscular junction (NMJ) in wild caught D. melanogaster as well as in over 20 other species of Drosophila. Whereas variation in NMJ morphology within a species is limited, I discovered a surprisingly extensive variation among different species. Compared with evolution of other morphological traits, NMJ morphology appears to be evolving very rapidly. Moreover, my data indicate that natural selection rather than genetic drift is primarily responsible for evolution of NMJ morphology. To dissect underlying molecular mechanisms that may govern NMJ growth and evolutionary divergence, I focused on a naturally occurring variant in D. melanogaster that causes NMJ overgrowth. I discovered that the variant mapped to Mob2, a gene encoding a kinase adapter protein originally described in yeast as a member of the Mitotic Exit Network (MEN). I have subsequently examined mutations in the Drosophila orthologs of all the core components of the yeast MEN and found that all of them function as part of a common pathway that acts presynaptically to negatively regulate NMJ growth. As in the regulation of yeast cytokinesis, these components of the MEN appear to act ultimately by regulating actin dynamics during the process of bouton growth and division. These studies have thus led to the discovery of an entirely new role for the MEN---regulation of synaptic growth---that is separate from its function in cell division. This work

  11. MicroRNA-dependent roles of Drosha and Pasha in the Drosophila larval ovary morphogenesis.

    PubMed

    Yang, Huiming; Li, Mengjie; Hu, Xiaolong; Xin, Tianchi; Zhang, Shu; Zhao, Gengchun; Xuan, Tao; Li, Mingfa

    2016-08-15

    The Drosophila larval ovary morphogenesis mainly involves coordinated development of somatic and germ cell lineages that is essential for forming a correct number of niche-germline stem cell (GSC) units (ovarioles) in the adult ovary. Ecdysone, Insulin, Activin, Dpp and EGFR signaling pathways form a regulatory network that orchestrates ovarian soma and germ line throughout larval development. Identification and characterization of additional genes or machineries involved in this process will provide more insights into the underlying mechanisms. Here, we show that the core microRNA (miRNA) pathway components Drosha and Pasha are required for coordinated development of somatic and germ cell precursors in the larval ovary. Drosha or pasha mutants display defective proliferation of primordial germ cells (PGCs), the precursors of GSCs prior to late third larval instar (LL3) and promoted PGC differentiation at LL3. In the mean time, loss of Drosha or Pasha function perturbs somatic precursor development, causing defects in formation of terminal filaments (TFs), a major composition of the GSC niche at LL3, as well as in TF precursor accumulation at early larval stages. Comparative analysis of the mutant phenotypes reveals that three other key miRNA pathway components, Dicer-1 (Dcr-1), Loquacious (Loqs) and Argonaute-1 (Ago-1) have similar effects as Drosha and Pasha indicated above, suggesting a role of the canonical miRNA pathway in the ovary development. Furthermore, genome-wide screening and genetic studies identify a set of Drosha-controlled miRNAs including miR-8, miR-14, miR-33, miR-184, miR-317 and let-7-C that function in this gonadogenesis. Taken together, this study provides the first ever demonstration that miRNA-mediated regulation is involved in the Drosophila larval ovary morphogenesis. PMID:27339292

  12. Functional genomics identifies regulators of the phototransduction machinery in the Drosophila larval eye and adult ocelli.

    PubMed

    Mishra, Abhishek Kumar; Bargmann, Bastiaan O R; Tsachaki, Maria; Fritsch, Cornelia; Sprecher, Simon G

    2016-02-15

    Sensory perception of light is mediated by specialized Photoreceptor neurons (PRs) in the eye. During development all PRs are genetically determined to express a specific Rhodopsin (Rh) gene and genes mediating a functional phototransduction pathway. While the genetic and molecular mechanisms of PR development is well described in the adult compound eye, it remains unclear how the expression of Rhodopsins and the phototransduction cascade is regulated in other visual organs in Drosophila, such as the larval eye and adult ocelli. Using transcriptome analysis of larval PR-subtypes and ocellar PRs we identify and study new regulators required during PR differentiation or necessary for the expression of specific signaling molecules of the functional phototransduction pathway. We found that the transcription factor Krüppel (Kr) is enriched in the larval eye and controls PR differentiation by promoting Rh5 and Rh6 expression. We also identified Camta, Lola, Dve and Hazy as key genes acting during ocellar PR differentiation. Further we show that these transcriptional regulators control gene expression of the phototransduction cascade in both larval eye and adult ocelli. Our results show that PR cell type-specific transcriptome profiling is a powerful tool to identify key transcriptional regulators involved during several aspects of PR development and differentiation. Our findings greatly contribute to the understanding of how combinatorial action of key transcriptional regulators control PR development and the regulation of a functional phototransduction pathway in both larval eye and adult ocelli. PMID:26769100

  13. Drosophila E-cadherin and its binding partner Armadillo/ beta-catenin are required for axonal pathway choices in the developing larval brain.

    PubMed

    Fung, Siaumin; Wang, Fay; Spindler, Shana R; Hartenstein, Volker

    2009-08-15

    The fly brain is formed by approximately hundred paired lineages of neurons, each lineage derived from one neuroblast. Embryonic neuroblasts undergo a small number of divisions and produce the primary neurons that form the functioning larval brain. In the larva, neuroblasts produce the secondary lineages that make up the bulk of the adult brain. Axons of a given secondary lineage fasciculate with each other and form a discrete bundle, the secondary axon tract (SAT). Secondary axon tracts prefigure the long axon connections of the adult brain, and therefore pathway choices of SATs made in the larva determine adult brain circuitry. Drosophila Shotgun/E-cadherin (DE-cad) and its binding partner Armadillo/beta-catenin (beta-cat) are expressed in newly born secondary neurons and their axons. The fact that the highly diverse, yet invariant pattern of secondary lineages and SATs has been recently mapped in the wild-type brain enabled us to investigate the role of DE-cad and beta-cat with the help of MARCM clones. Clones were validated by their absence of DE-cad immuno-reactivity. The most significant phenotype consists in the defasciculation and an increased amount of branching of SATs at the neuropile-cortex boundary, as well as subtle changes in the trajectory of SATs within the neuropile. In general, only a fraction of mutant clones in a given lineage showed structural abnormalities. Furthermore, although they all globally express DE-cad and beta-cat, lineages differ in their requirement for DE-cad function. Some lineages never showed morphological abnormalities in MARCM clones, whereas others reacted with abnormal branching and changes in SAT trajectory at a high frequency. We conclude that DE-cad/beta-cat form part of the mechanism that control branching and trajectory of axon tracts in the larval brain. PMID:19520071

  14. Drosophila E-cadherin and its binding partner Armadillo/ β-catenin are required for axonal pathway choices in the developing larval brain

    PubMed Central

    Fung, Siaumin; Wang, Fay; Spindler, Shana R; Hartenstein, Volker

    2009-01-01

    The fly brain is formed by approximately hundred paired lineages of neurons, each lineage derived from one neuroblast. Embryonic neuroblasts undergo a small number of divisions and produce the primary neurons that form the functioning larval brain. In the larva, neuroblasts produce the secondary lineages that make up the bulk of the adult brain. Axons of a given secondary lineage fasciculate with each other and form a discrete bundle, the secondary axon tract (SAT). Secondary axon tracts prefigure the long axon connections of the adult brain, and therefore pathway choices of SATs made in the larva determine adult brain circuitry. Drosophila Shotgun/E-cadherin (DE-cad) and its binding partner Armadillo/β-catenin (β-cat) are expressed in newly born secondary neurons and their axons. The fact that the highly diverse, yet invariant pattern of secondary lineages and SATs has been recently mapped in the wild-type brain enabled us to investigate the role of DE-cad and β-cat with the help of MARCM clones. Clones were validated by their absence of DE-cad immuno-reactivity. The most significant phenotype consists in the defasciculation and an increased amount of branching of SATs at the neuropile-cortex boundary, as well as subtle changes in the trajectory of SATs within the neuropile. In general, only a fraction of mutant clones in a given lineage showed structural abnormalities. Furthermore, although they all globally express DE-cad and β-cat, lineages differ in their requirement for DE-cad function. Some lineages never showed morphological abnormalities in MARCM clones, whereas others reacted with abnormal branching and changes in SAT trajectory at a high frequency. We conclude that DE-cad/β-cat form part of the mechanism that control branching and trajectory of axon tracts in the larval brain. PMID:19520071

  15. Properties of the larval neuromuscular junction in Drosophila melanogaster.

    PubMed Central

    Jan, L Y; Jan, Y N

    1976-01-01

    The anatomy and physiology of the Drosophila larval neuromuscular junction were studied. 2. The dependence of muscle resting potentials on [K+]o and [Na+]o follows the Goldman-Hodgkin-Katz equation (PNa/PK=0-23). Chloride ions distribute passively across the membrane. 3. The mean specific membrane resistance of muscle fibres is 4-3 X 10(3) omega cm2, and the mean specific membrane capacitance is 7-1 muF/cm2. The muscle fibre is virtually isopotential. 4.Transmitter release is quantal. Both the miniature excitatory junctional potential and the evoked release follow the Poisson distribution. 5. Transmitter release depends on approximately the fourth power of [Ca2+]o. If Sr2+ replaces Ca2+, it depends on approximately the fourth power of [Sr2+]o. Mg2+ reduces transmitter release without altering the fourth power dependence on [Ca2+]o. Images A B A B Plate 3 PMID:11339

  16. The Role of Dopamine in Drosophila Larval Classical Olfactory Conditioning

    PubMed Central

    Han, Kyung-An; Stocker, Reinhard F.; Thum, Andreas S.

    2009-01-01

    Learning and memory is not an attribute of higher animals. Even Drosophila larvae are able to form and recall an association of a given odor with an aversive or appetitive gustatory reinforcer. As the Drosophila larva has turned into a particularly simple model for studying odor processing, a detailed neuronal and functional map of the olfactory pathway is available up to the third order neurons in the mushroom bodies. At this point, a convergence of olfactory processing and gustatory reinforcement is suggested to underlie associative memory formation. The dopaminergic system was shown to be involved in mammalian and insect olfactory conditioning. To analyze the anatomy and function of the larval dopaminergic system, we first characterize dopaminergic neurons immunohistochemically up to the single cell level and subsequent test for the effects of distortions in the dopamine system upon aversive (odor-salt) as well as appetitive (odor-sugar) associative learning. Single cell analysis suggests that dopaminergic neurons do not directly connect gustatory input in the larval suboesophageal ganglion to olfactory information in the mushroom bodies. However, a number of dopaminergic neurons innervate different regions of the brain, including protocerebra, mushroom bodies and suboesophageal ganglion. We found that dopamine receptors are highly enriched in the mushroom bodies and that aversive and appetitive olfactory learning is strongly impaired in dopamine receptor mutants. Genetically interfering with dopaminergic signaling supports this finding, although our data do not exclude on naïve odor and sugar preferences of the larvae. Our data suggest that dopaminergic neurons provide input to different brain regions including protocerebra, suboesophageal ganglion and mushroom bodies by more than one route. We therefore propose that different types of dopaminergic neurons might be involved in different types of signaling necessary for aversive and appetitive olfactory memory

  17. Sonication-facilitated Immunofluorescence Staining of Late-stage Embryonic and Larval Drosophila Tissues In Situ

    PubMed Central

    Wawersik, Matthew

    2014-01-01

    Studies performed in Drosophila melanogaster embryos and larvae provide crucial insight into developmental processes such as cell fate specification and organogenesis. Immunostaining allows for the visualization of developing tissues and organs. However, a protective cuticle that forms at the end of embryogenesis prevents permeation of antibodies into late-stage embryos and larvae. While dissection prior to immunostaining is regularly used to analyze Drosophila larval tissues, it proves inefficient for some analyses because small tissues may be difficult to locate and isolate. Sonication provides an alternative to dissection in larval Drosophila immunostaining protocols. It allows for quick, simultaneous processing of large numbers of late-stage embryos and larvae and maintains in situ morphology. After fixation in formaldehyde, a sample is sonicated. Sample is then subjected to immunostaining with antigen-specific primary antibodies and fluorescently labeled secondary antibodies to visualize target cell types and specific proteins via fluorescence microscopy. During the process of sonication, proper placement of a sonicating probe above the sample, as well as the duration and intensity of sonication, is critical. Additonal minor modifications to standard immunostaining protocols may be required for high quality stains. For antibodies with low signal to noise ratio, longer incubation times are typically necessary. As a proof of concept for this sonication-facilitated protocol, we show immunostains of three tissue types (testes, ovaries, and neural tissues) at a range of developmental stages. PMID:25146311

  18. Genetic analysis of Eclosion hormone action during Drosophila larval ecdysis.

    PubMed

    Krüger, Eileen; Mena, Wilson; Lahr, Eleanor C; Johnson, Erik C; Ewer, John

    2015-12-15

    Insect growth is punctuated by molts, during which the animal produces a new exoskeleton. The molt culminates in ecdysis, an ordered sequence of behaviors that causes the old cuticle to be shed. This sequence is activated by Ecdysis triggering hormone (ETH), which acts on the CNS to activate neurons that produce neuropeptides implicated in ecdysis, including Eclosion hormone (EH), Crustacean cardioactive peptide (CCAP) and Bursicon. Despite more than 40 years of research on ecdysis, our understanding of the precise roles of these neurohormones remains rudimentary. Of particular interest is EH; although it is known to upregulate ETH release, other roles for EH have remained elusive. We isolated an Eh null mutant in Drosophila and used it to investigate the role of EH in larval ecdysis. We found that null mutant animals invariably died at around the time of ecdysis, revealing an essential role in its control. Further analyses showed that these animals failed to express the preparatory behavior of pre-ecdysis while directly expressing the motor program of ecdysis. Although ETH release could not be detected, the lack of pre-ecdysis could not be rescued by injections of ETH, suggesting that EH is required within the CNS for ETH to trigger the normal ecdysial sequence. Using a genetically encoded calcium probe, we showed that EH configured the response of the CNS to ETH. These findings show that EH plays an essential role in the Drosophila CNS in the control of ecdysis, in addition to its known role in the periphery of triggering ETH release. PMID:26395475

  19. Whole-central nervous system functional imaging in larval Drosophila

    PubMed Central

    Lemon, William C.; Pulver, Stefan R.; Höckendorf, Burkhard; McDole, Katie; Branson, Kristin; Freeman, Jeremy; Keller, Philipp J.

    2015-01-01

    Understanding how the brain works in tight concert with the rest of the central nervous system (CNS) hinges upon knowledge of coordinated activity patterns across the whole CNS. We present a method for measuring activity in an entire, non-transparent CNS with high spatiotemporal resolution. We combine a light-sheet microscope capable of simultaneous multi-view imaging at volumetric speeds 25-fold faster than the state-of-the-art, a whole-CNS imaging assay for the isolated Drosophila larval CNS and a computational framework for analysing multi-view, whole-CNS calcium imaging data. We image both brain and ventral nerve cord, covering the entire CNS at 2 or 5 Hz with two- or one-photon excitation, respectively. By mapping network activity during fictive behaviours and quantitatively comparing high-resolution whole-CNS activity maps across individuals, we predict functional connections between CNS regions and reveal neurons in the brain that identify type and temporal state of motor programs executed in the ventral nerve cord. PMID:26263051

  20. Using en-face optical coherence tomography to analyse gene function in Drosophila Melanogaster larval heart

    NASA Astrophysics Data System (ADS)

    Bradu, Adrian; Ma, Lisha; Bloor, Jim; Podoleanu, Adrian

    2008-09-01

    In-vivo Optical Coherence Tomography (OCT) imaging of the fruit fly Drosophila Melanogaster larval heart allows non invasive visualizations and assessment of its cardiac function. In order to image Drosophila heart, we have developed a dedicated imaging instrument able to provide simultaneous OCT and Laser Scanning Confocal Microscopy (LSCM) images. With this dual imaging system, the heart can easily be located and visualised within the specimen and the change of the heart shape in a cardiac cycle monitored. Here we have used targeted gene expression to knockdown the myospheroid (mys) gene in the larval heart using a specific RNAi construct. By knocking down a β integrin subunit encoded by mys we have recorded an enlarged heart chamber in both diastolic and systolic states. Also, the fraction of reduction of the chamber diameter was smaller in the knockdown heart. These phenotypic differences indicate that impaired cardiac contractility occurs in the heart where the integrin gene express level is reduced. At our knowledge, this is for the first time when it is shown that integrins have a direct relationship to a dilated heart defect.

  1. Sequential en-face optical coherence tomography imaging and monitoring of Drosophila Melanogaster larval heart

    NASA Astrophysics Data System (ADS)

    Bradu, A.; Ma, Lisha; Bloor, J.; Podoleanu, A. GH.

    2009-02-01

    This article demonstrates two modalities to acquire information on cardiac function in larval Drosophila Melanogaster: in-vivo imaging and heartbeat monitoring. To achieve these goals a dedicated imaging instrument able to provide simultaneous en-face Optical Coherence Tomography (OCT) and Laser Scanning Confocal Microscopy (LSCM) images has been developed. With this dual imaging system, the heart can easily be located and visualised within the specimen and the change of the heart shape in a cardiac cycle monitored. The system can easily be switched to a stethoscopic regime, simply by interrupting the scanning of the light beam across the sample, after selecting the point of interest in the imaging regime. Here we have used targeted gene expression to knockdown the myospheroid (mys) gene in the larval heart using a specific RNAi construct. By knocking down a β integrin subunit encoded by mys we have recorded an enlarged heart chamber in both diastolic and systolic states. Also, the fraction of reduction of the chamber diameter was smaller in the knockdown heart. These phenotypic differences indicate that impaired cardiac contractility occurs in the heart where the integrin gene express level is reduced. As far as we are aware, this is for the first time when it is shown in Drosophila that integrins have a direct relationship to a dilated heart defect, and conseqThis article demonstrates two modalities to acquire information on cardiac function in larval Drosophila Melanogaster: in-vivo imaging and heartbeat monitoring. To achieve these goals a dedicated imaging instrument able to provide simultaneous en-face Optical Coherence Tomography (OCT) and Laser Scanning Confocal Microscopy (LSCM) images has been developed. With this dual imaging system, the heart can easily be located and visualised within the specimen and the change of the heart shape in a cardiac cycle monitored. The system can easily be switched to a stethoscopic regime, simply by interrupting the

  2. Evolution of increased adult longevity in Drosophila melanogaster populations selected for adaptation to larval crowding.

    PubMed

    Shenoi, V N; Ali, S Z; Prasad, N G

    2016-02-01

    In holometabolous animals such as Drosophila melanogaster, larval crowding can affect a wide range of larval and adult traits. Adults emerging from high larval density cultures have smaller body size and increased mean life span compared to flies emerging from low larval density cultures. Therefore, adaptation to larval crowding could potentially affect adult longevity as a correlated response. We addressed this issue by studying a set of large, outbred populations of D. melanogaster, experimentally evolved for adaptation to larval crowding for 83 generations. We assayed longevity of adult flies from both selected (MCUs) and control populations (MBs) after growing them at different larval densities. We found that MCUs have evolved increased mean longevity compared to MBs at all larval densities. The interaction between selection regime and larval density was not significant, indicating that the density dependence of mean longevity had not evolved in the MCU populations. The increase in longevity in MCUs can be partially attributed to their lower rates of ageing. It is also noteworthy that reaction norm of dry body weight, a trait probably under direct selection in our populations, has indeed evolved in MCU populations. To the best of our knowledge, this is the first report of the evolution of adult longevity as a correlated response of adaptation to larval crowding. PMID:26575793

  3. 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...

  4. Natural variability in Drosophila larval and pupal NaCl tolerance.

    PubMed

    Riedl, Craig A L; Oster, Sara; Busto, Macarena; Mackay, Trudy F C; Sokolowski, Marla B

    2016-05-01

    The regulation of NaCl is essential for the maintenance of cellular tonicity and functionality, and excessive salt exposure has many adverse effects. The fruit fly, Drosophila melanogaster, is a good osmoregulator and some strains can survive on media with very low or high NaCl content. Previous analyses of mutant alleles have implicated various stress signaling cascades in NaCl sensitivity or tolerance; however, the genes influencing natural variability of NaCl tolerance remain for the most part unknown. Here, we use two approaches to investigate natural variation in D. melanogaster NaCl tolerance. We describe four D. melanogaster lines that were selected for different degrees of NaCl tolerance, and present data on their survival, development, and pupation position when raised on varying NaCl concentrations. After finding evidence for natural variation in salt tolerance, we present the results of Quantitative Trait Loci (QTL) mapping of natural variation in larval and pupal NaCl tolerance, and identify different genomic regions associated with NaCl tolerance during larval and pupal development. PMID:26874056

  5. Pharmacological identification of cholinergic receptor subtypes on Drosophila melanogaster larval heart.

    PubMed

    Malloy, Cole A; Ritter, Kyle; Robinson, Jonathan; English, Connor; Cooper, Robin L

    2016-01-01

    The Drosophila melanogaster heart is a popular model in which to study cardiac physiology and development. Progress has been made in understanding the role of endogenous compounds in regulating cardiac function in this model. It is well characterized that common neurotransmitters act on many peripheral and non-neuronal tissues as they flow through the hemolymph of insects. Many of these neuromodulators, including acetylcholine (ACh), have been shown to act directly on the D. melanogaster larval heart. ACh is a primary neurotransmitter in the central nervous system (CNS) of vertebrates and at the neuromuscular junctions on skeletal and cardiac tissue. In insects, ACh is the primary excitatory neurotransmitter of sensory neurons and is also prominent in the CNS. A full understanding regarding the regulation of the Drosophila cardiac physiology by the cholinergic system remains poorly understood. Here we use semi-intact D. melanogaster larvae to study the pharmacological profile of cholinergic receptor subtypes, nicotinic acetylcholine receptors (nAChRs) and muscarinic acetylcholine receptors (mAChRs), in modulating heart rate (HR). Cholinergic receptor agonists, nicotine and muscarine both increase HR, while nAChR agonist clothianidin exhibits no significant effect when exposed to an open preparation at concentrations as low as 100 nM. In addition, both nAChR and mAChR antagonists increase HR as well but also display capabilities of blocking agonist actions. These results provide evidence that both of these receptor subtypes display functional significance in regulating the larval heart's pacemaker activity. PMID:26438517

  6. Larval Population Density Alters Adult Sleep in Wild-Type Drosophila melanogaster but Not in Amnesiac Mutant Flies

    PubMed Central

    Chi, Michael W.; Griffith, Leslie C.; Vecsey, Christopher G.

    2014-01-01

    Sleep has many important biological functions, but how sleep is regulated remains poorly understood. In humans, social isolation and other stressors early in life can disrupt adult sleep. In fruit flies housed at different population densities during early adulthood, social enrichment was shown to increase subsequent sleep, but it is unknown if population density during early development can also influence adult sleep. To answer this question, we maintained Drosophila larvae at a range of population densities throughout larval development, kept them isolated during early adulthood, and then tested their sleep patterns. Our findings reveal that flies that had been isolated as larvae had more fragmented sleep than those that had been raised at higher population densities. This effect was more prominent in females than in males. Larval population density did not affect sleep in female flies that were mutant for amnesiac, which has been shown to be required for normal memory consolidation, adult sleep regulation, and brain development. In contrast, larval population density effects on sleep persisted in female flies lacking the olfactory receptor or83b, suggesting that olfactory signals are not required for the effects of larval population density on adult sleep. These findings show that population density during early development can alter sleep behavior in adulthood, suggesting that genetic and/or structural changes are induced by this developmental manipulation that persist through metamorphosis. PMID:25116571

  7. Drosophila Food-Associated Pheromones: Effect of Experience, Genotype and Antibiotics on Larval Behavior

    PubMed Central

    Thibert, Julien; Farine, Jean-Pierre; Cortot, Jérôme; Ferveur, Jean-François

    2016-01-01

    Animals ubiquitously use chemical signals to communicate many aspects of their social life. These chemical signals often consist of environmental cues mixed with species-specific signals—pheromones—emitted by conspecifics. During their life, insects can use pheromones to aggregate, disperse, choose a mate, or find the most suitable food source on which to lay eggs. Before pupariation, larvae of several Drosophila species migrate to food sources depending on their composition and the presence of pheromones. Some pheromones derive from microbiota gut activity and these food-associated cues can enhance larval attraction or repulsion. To explore the mechanisms underlying the preference (attraction/repulsion) to these cues and clarify their effect, we manipulated factors potentially involved in larval response. In particular, we found that the (i) early exposure to conspecifics, (ii) genotype, and (iii) antibiotic treatment changed D. melanogaster larval behavior. Generally, larvae—tested either individually or in groups—strongly avoided food processed by other larvae. Compared to previous reports on larval attractive pheromones, our data suggest that such attractive effects are largely masked by food-associated compounds eliciting larval aversion. The antagonistic effect of attractive vs. aversive compounds could modulate larval choice of a pupariation site and impact the dispersion of individuals in nature. PMID:26987117

  8. Insulin- and warts-dependent regulation of tracheal plasticity modulates systemic larval growth during hypoxia in Drosophila melanogaster.

    PubMed

    Wong, Daniel M; Shen, Zhouyang; Owyang, Kristin E; Martinez-Agosto, Julian A

    2014-01-01

    Adaptation to dynamic environmental cues during organismal development requires coordination of tissue growth with available resources. More specifically, the effects of oxygen availability on body size have been well-documented, but the mechanisms through which hypoxia restricts systemic growth have not been fully elucidated. Here, we characterize the larval growth and metabolic defects in Drosophila that result from hypoxia. Hypoxic conditions reduced fat body opacity and increased lipid droplet accumulation in this tissue, without eliciting lipid aggregation in hepatocyte-like cells called oenocytes. Additionally, hypoxia increased the retention of Dilp2 in the insulin-producing cells of the larval brain, associated with a reduction of insulin signaling in peripheral tissues. Overexpression of the wildtype form of the insulin receptor ubiquitously and in the larval trachea rendered larvae resistant to hypoxia-induced growth restriction. Furthermore, Warts downregulation in the trachea was similar to increased insulin receptor signaling during oxygen deprivation, which both rescued hypoxia-induced growth restriction, inhibition of tracheal molting, and developmental delay. Insulin signaling and loss of Warts function increased tracheal growth and augmented tracheal plasticity under hypoxic conditions, enhancing oxygen delivery during periods of oxygen deprivation. Our findings demonstrate a mechanism that coordinates oxygen availability with systemic growth in which hypoxia-induced reduction of insulin receptor signaling decreases plasticity of the larval trachea that is required for the maintenance of systemic growth during times of limiting oxygen availability. PMID:25541690

  9. Insulin- and Warts-Dependent Regulation of Tracheal Plasticity Modulates Systemic Larval Growth during Hypoxia in Drosophila melanogaster

    PubMed Central

    Wong, Daniel M.; Shen, Zhouyang; Owyang, Kristin E.; Martinez-Agosto, Julian A.

    2014-01-01

    Adaptation to dynamic environmental cues during organismal development requires coordination of tissue growth with available resources. More specifically, the effects of oxygen availability on body size have been well-documented, but the mechanisms through which hypoxia restricts systemic growth have not been fully elucidated. Here, we characterize the larval growth and metabolic defects in Drosophila that result from hypoxia. Hypoxic conditions reduced fat body opacity and increased lipid droplet accumulation in this tissue, without eliciting lipid aggregation in hepatocyte-like cells called oenocytes. Additionally, hypoxia increased the retention of Dilp2 in the insulin-producing cells of the larval brain, associated with a reduction of insulin signaling in peripheral tissues. Overexpression of the wildtype form of the insulin receptor ubiquitously and in the larval trachea rendered larvae resistant to hypoxia-induced growth restriction. Furthermore, Warts downregulation in the trachea was similar to increased insulin receptor signaling during oxygen deprivation, which both rescued hypoxia-induced growth restriction, inhibition of tracheal molting, and developmental delay. Insulin signaling and loss of Warts function increased tracheal growth and augmented tracheal plasticity under hypoxic conditions, enhancing oxygen delivery during periods of oxygen deprivation. Our findings demonstrate a mechanism that coordinates oxygen availability with systemic growth in which hypoxia-induced reduction of insulin receptor signaling decreases plasticity of the larval trachea that is required for the maintenance of systemic growth during times of limiting oxygen availability. PMID:25541690

  10. Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction.

    PubMed

    Sanhueza, Mario; Kubasik-Thayil, Anisha; Pennetta, Giuseppa

    2016-01-01

    Most studies on morphogenesis rely on qualitative descriptions of how anatomical traits are affected by the disruption of specific genes and genetic pathways. Quantitative descriptions are rarely performed, although genetic manipulations produce a range of phenotypic effects and variations are observed even among individuals within control groups. Emerging evidence shows that morphology, size and location of organelles play a previously underappreciated, yet fundamental role in cell function and survival. Here we provide step-by-step instructions for performing quantitative analyses of phenotypes at the Drosophila larval neuromuscular junction (NMJ). We use several reliable immuno-histochemical markers combined with bio-imaging techniques and morphometric analyses to examine the effects of genetic mutations on specific cellular processes. In particular, we focus on the quantitative analysis of phenotypes affecting morphology, size and position of nuclei within the striated muscles of Drosophila larvae. The Drosophila larval NMJ is a valuable experimental model to investigate the molecular mechanisms underlying the structure and the function of the neuromuscular system, both in health and disease. However, the methodologies we describe here can be extended to other systems as well. PMID:27213489

  11. Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

    PubMed Central

    Pennetta, Giuseppa

    2016-01-01

    Most studies on morphogenesis rely on qualitative descriptions of how anatomical traits are affected by the disruption of specific genes and genetic pathways. Quantitative descriptions are rarely performed, although genetic manipulations produce a range of phenotypic effects and variations are observed even among individuals within control groups. Emerging evidence shows that morphology, size and location of organelles play a previously underappreciated, yet fundamental role in cell function and survival. Here we provide step-by-step instructions for performing quantitative analyses of phenotypes at the Drosophila larval neuromuscular junction (NMJ). We use several reliable immuno-histochemical markers combined with bio-imaging techniques and morphometric analyses to examine the effects of genetic mutations on specific cellular processes. In particular, we focus on the quantitative analysis of phenotypes affecting morphology, size and position of nuclei within the striated muscles of Drosophila larvae. The Drosophila larval NMJ is a valuable experimental model to investigate the molecular mechanisms underlying the structure and the function of the neuromuscular system, both in health and disease. However, the methodologies we describe here can be extended to other systems as well. PMID:27213489

  12. Associative learning between odorants and mechanosensory punishment in larval Drosophila.

    PubMed

    Eschbach, Claire; Cano, Carmen; Haberkern, Hannah; Schraut, Karla; Guan, Chonglin; Triphan, Tilman; Gerber, Bertram

    2011-12-01

    We tested whether Drosophila larvae can associate odours with a mechanosensory disturbance as a punishment, using substrate vibration conveyed by a loudspeaker (buzz:). One odour (A) was presented with the buzz, while another odour (B) was presented without the buzz (A/B training). Then, animals were offered the choice between A and B. After reciprocal training (A/B), a second experimental group was tested in the same way. We found that larvae show conditioned escape from the previously punished odour. We further report an increase of associative performance scores with the number of punishments, and an increase according to the number of training cycles. Within the range tested (between 50 and 200 Hz), however, the pitch of the buzz does not apparently impact associative success. Last, but not least, we characterized odour-buzz memories with regard to the conditions under which they are behaviourally expressed--or not. In accordance with what has previously been found for associative learning between odours and bad taste (such as high concentration salt or quinine), we report that conditioned escape after odour-buzz learning is disabled if escape is not warranted, i.e. if no punishment to escape from is present during testing. Together with the already established paradigms for the association of odour and bad taste, the present assay offers the prospect of analysing how a relatively simple brain orchestrates memory and behaviour with regard to different kinds of 'bad' events. PMID:22071180

  13. Developmental compartments in the larval trachea of Drosophila

    PubMed Central

    Rao, Prashanth R; Lin, Li; Huang, Hai; Guha, Arjun; Roy, Sougata; Kornberg, Thomas B

    2015-01-01

    The Drosophila tracheal system is a branched tubular network that forms in the embryo by a post-mitotic program of morphogenesis. In third instar larvae (L3), cells constituting the second tracheal metamere (Tr2) reenter the cell cycle. Clonal analysis of L3 Tr2 revealed that dividing cells in the dorsal trunk, dorsal branch and transverse connective branches respect lineage restriction boundaries near branch junctions. These boundaries corresponded to domains of gene expression, for example where cells expressing Spalt, Delta and Serrate in the dorsal trunk meet vein–expressing cells in the dorsal branch or transverse connective. Notch signaling was activated to one side of these borders and was required for the identity, specializations and segregation of border cells. These findings suggest that Tr2 is comprised of developmental compartments and that developmental compartments are an organizational feature relevant to branched tubular networks. DOI: http://dx.doi.org/10.7554/eLife.08666.001 PMID:26491942

  14. Failure to Burrow and Tunnel Reveals Roles for jim lovell in the Growth and Endoreplication of the Drosophila Larval Tracheae

    PubMed Central

    Qiang, Karen M.; Beckingham, Kathleen M.

    2016-01-01

    The Drosophila protein Jim Lovell (Lov) is a putative transcription factor of the BTB/POZ (Bric- a-Brac/Tramtrack/Broad/ Pox virus and Zinc finger) domain class that is expressed in many elements of the developing larval nervous system. It has roles in innate behaviors such as larval locomotion and adult courtship. In performing tissue-specific knockdown with the Gal4-UAS system we identified a new behavioral phenotype for lov: larvae failed to burrow into their food during their growth phase and then failed to tunnel into an agarose substratum during their wandering phase. We determined that these phenotypes originate in a previously unrecognized role for lov in the tracheae. By using tracheal-specific Gal4 lines, Lov immunolocalization and a lov enhancer trap line, we established that lov is normally expressed in the tracheae from late in embryogenesis through larval life. Using an assay that monitors food burrowing, substrate tunneling and death we showed that lov tracheal knockdown results in tracheal fluid-filling, producing hypoxia that activates the aberrant behaviors and inhibits development. We investigated the role of lov in the tracheae that initiates this sequence of events. We discovered that when lov levels are reduced, the tracheal cells are smaller, more numerous and show lower levels of endopolyploidization. Together our findings indicate that Lov is necessary for tracheal endoreplicative growth and that its loss in this tissue causes loss of tracheal integrity resulting in chronic hypoxia and abnormal burrowing and tunneling behavior. PMID:27494251

  15. Temperature and parasitism by Asobara tabida (Hymenoptera: Braconidae) influence larval pupation behaviour in two Drosophila species

    NASA Astrophysics Data System (ADS)

    Josso, Céline; Moiroux, Joffrey; Vernon, Philippe; van Baaren, Joan; van Alphen, Jacques J. M.

    2011-08-01

    In holometabolous insects, pupation site selection behaviour has large consequences for survival. Here, we investigated the combined effects of temperature and parasitism by the parasitoid Asobara tabida on larval pupation behaviour in two of its main Drosophila sp. hosts differing in their climate origin. We found that larvae of Drosophila melanogaster—a species with a (sub)tropical origin—placed at 25°C pupated higher in rearing jars than those placed at 15°C. The opposite pattern was observed for Drosophila subobscura larvae—a species from temperate regions—which pupated lower, i.e. on or near the substrate at 25°C, than those placed at 15°C. When placed at 25°C, parasitized larvae of both species pupated closer to the substrate than unparasitized ones. Moreover, the Drosophila larvae that had been exposed and probably stung by A. tabida, but were not parasitized, pupated lower than the control unparasitized larvae. These results provide new insights of host behaviour manipulation by A. tabida larvae.

  16. Neuroblast lineage-specific origin of the neurons of the Drosophila larval olfactory system

    PubMed Central

    Das, Abhijit; Gupta, Tripti; Davla, Sejal; Godino, Laura Lucia Prieto; Diegelmann, Sören; Reddy, O. Venkateswara; VijayRaghavan, K.; Reichert, Heinrich; Lovick, Jennifer; Hartenstein, Volker

    2014-01-01

    The complete neuronal repertoire of the central brain of Drosophila originates from only approximately 100 pairs of neural stem cells, or neuroblasts. Each neuroblast produces a highly stereotyped lineage of neurons which innervate specific compartments of the brain. Neuroblasts undergo two rounds of mitotic activity: embryonic divisions produce lineages of primary neurons that build the larval nervous system; after a brief quiescence, the neuroblasts go through a second round of divisions in larval stage to produce secondary neurons which are integrated into the adult nervous system. Here we investigate the lineages that are associated with the larval antennal lobe, one of the most widely studied neuronal systems in fly. We find that the same five neuroblasts responsible for the adult antennal lobe also produce the antennal lobe of the larval brain. However, there are notable differences in the composition of larval (primary) lineages and their adult (secondary) counterparts. Significantly, in the adult, two lineages (lNB/BAlc and adNB/BAmv3) produce uniglomerular projection neurons connecting the antennal lobe with the mushroom body and lateral horn; another lineage, vNB/BAla1, generates multiglomerular neurons reaching the lateral horn directly. lNB/BAlc, as well as a fourth lineage, vlNB/BAla2, generate a diversity of local interneurons. We describe a fifth, previously unknown lineage, BAlp4, which connects the posterior part of the antennal lobe and the neighboring tritocerebrum (gustatory center) with a higher brain center located adjacent to the mushroom body. In the larva, only one of these lineages, adNB/BAmv3, generates all uniglomerular projection neurons. Also as in the adult, lNB/BAlc and vlNB/BAla2 produce local interneurons which, in terms of diversity in architecture and transmitter expression, resemble their adult counterparts. In addition, lineages lNB/BAlc and vNB/BAla1, as well as the newly described BAlp4, form numerous types of projection

  17. Neuroblast lineage-specific origin of the neurons of the Drosophila larval olfactory system.

    PubMed

    Das, Abhijit; Gupta, Tripti; Davla, Sejal; Prieto-Godino, Lucia L; Diegelmann, Sören; Reddy, O Venkateswara; Raghavan, K Vijay; Reichert, Heinrich; Lovick, Jennifer; Hartenstein, Volker

    2013-01-15

    The complete neuronal repertoire of the central brain of Drosophila originates from only approximately 100 pairs of neural stem cells, or neuroblasts. Each neuroblast produces a highly stereotyped lineage of neurons which innervate specific compartments of the brain. Neuroblasts undergo two rounds of mitotic activity: embryonic divisions produce lineages of primary neurons that build the larval nervous system; after a brief quiescence, the neuroblasts go through a second round of divisions in larval stage to produce secondary neurons which are integrated into the adult nervous system. Here we investigate the lineages that are associated with the larval antennal lobe, one of the most widely studied neuronal systems in fly. We find that the same five neuroblasts responsible for the adult antennal lobe also produce the antennal lobe of the larval brain. However, there are notable differences in the composition of larval (primary) lineages and their adult (secondary) counterparts. Significantly, in the adult, two lineages (lNB/BAlc and adNB/BAmv3) produce uniglomerular projection neurons connecting the antennal lobe with the mushroom body and lateral horn; another lineage, vNB/BAla1, generates multiglomerular neurons reaching the lateral horn directly. lNB/BAlc, as well as a fourth lineage, vlNB/BAla2, generate a diversity of local interneurons. We describe a fifth, previously unknown lineage, BAlp4, which connects the posterior part of the antennal lobe and the neighboring tritocerebrum (gustatory center) with a higher brain center located adjacent to the mushroom body. In the larva, only one of these lineages, adNB/BAmv3, generates all uniglomerular projection neurons. Also as in the adult, lNB/BAlc and vlNB/BAla2 produce local interneurons which, in terms of diversity in architecture and transmitter expression, resemble their adult counterparts. In addition, lineages lNB/BAlc and vNB/BAla1, as well as the newly described BAlp4, form numerous types of projection

  18. 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

  19. Drosulfakinin activates CCKLR-17D1 and promotes larval locomotion and escape response in Drosophila

    PubMed Central

    Chen, Xu; Peterson, Jonathan; Nachman, Ronald J.; Ganetzky, Barry

    2012-01-01

    Neuropeptides are ubiquitous in both mammals and invertebrates and play essential roles in regulation and modulation of many developmental and physiological processes through activation of G-protein-coupled-receptors (GPCRs). However, the mechanisms by which many of the neuropeptides regulate specific neural function and behaviors remain undefined. Here we investigate the functions of Drosulfakinin (DSK), the Drosophila homolog of vertebrate neuropeptide cholecystokinin (CCK), which is the most abundant neuropeptide in the central nervous system. We provide biochemical evidence that sulfated DSK-1 and DSK-2 activate the CCKLR-17D1 receptor in a cell culture assay. We further examine the role of DSK and CCKLR-17D1 in the regulation of larval locomotion, both in a semi-intact larval preparation and in intact larvae under intense light exposure. Our results suggest that DSK/CCKLR-17D1 signaling promote larval body wall muscle contraction and is necessary for mediating locomotor behavior in stress-induced escape response. PMID:22885328

  20. 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

  1. A reappraisal of the hormonal regulation of larval fat body histolysis in female Drosophila melanogaster.

    PubMed

    Richard, D S; Arnim, A E; Gilbert, L I

    1993-02-15

    The histolysis of larval fat body cells in adult female Drosophila melanogaster was examined in wild type and mutant animals. The fat body cells of wild type (Canton-S), apterous56f homozygotes, apterous78jts homozygotes and heterozygotes, apterous4/+, ecdysoneless1 homozygotes and heterozygotes all underwent histolysis normally during the 72 h following adult eclosion. Only in the case of ap4/ap4 adults did the cells fail to histolyze normally. The fat body cells of both diapausing and non-diapausing wild type females underwent histolysis at the same rate. Attempts to demonstrate histolysis in vitro were unsuccessful, even in the presence of juvenile hormones (JHs), larval ring glands, or adult ovaries. In all strains other than the ap4 homozygotes, a significant proportion of larval fat body cells were dead at any time while the ap4/ap4 animals, almost all cells remained viable. It is postulated that fat body cell lysis following eclosion is not a JH-mediated event, but is elicited by an as yet unidentified factor(s), possibly originating in the ovary. PMID:8440351

  2. Maintenance and regulation of extracellular volume and the ion environment in Drosophila larval nerves

    PubMed Central

    Leiserson, William M.; Keshishian, Haig

    2010-01-01

    In mammals and insects, paracellular blood barriers isolate the nervous system from the rest of the animal. Glia and accessory cells of the nervous system use pumps, channels, cotransporters, and exchangers collectively to maintain the extracellular ion environment and osmotic balance in the nervous system. At present the molecular mechanisms that regulate this process remain unclear. In humans, loss of extracellular ion and volume regulation in the nervous system poses serious health threats. Drosophila is a model genetic organism with a proven track record for uncovering molecular mechanisms relevant to human health and disease. Here, we review what is known about extracellular ion and volume regulation in larval abdominal nerves, present some new data about the impact of neural activity on the extracellular environment, and relate the findings to mammalian systems. Homologies have been found at the level of morphology, physiology, molecular mechanisms, and mutant phenotypes. The Fray-Ncc69 module regulates extracellular volume in larval nerves. Genetic rescue experiments with the mammalian orthologs prove this module has a direct correlate in humans. This and other molecular homologies, together with the similar physiological needs, suggest that uncovering the molecular mechanisms of ion and volume regulation in larval nerves will likely provide significant insights into this process in mammalian systems. PMID:21305613

  3. Drosophila 4EHP is essential for the larval-pupal transition and required in the prothoracic gland for ecdysone biosynthesis.

    PubMed

    Valzania, Luca; Ono, Hajime; Ignesti, Marilena; Cavaliere, Valeria; Bernardi, Fabio; Gamberi, Chiara; Lasko, Paul; Gargiulo, Giuseppe

    2016-02-01

    Maternal expression of the translational regulator 4EHP (eIF4E-Homologous Protein) has an established role in generating protein gradients essential for specifying the Drosophila embryonic pattern. We generated a null mutation of 4EHP, which revealed for the first time that it is essential for viability and for completion of development. In fact, 4EHP null larvae, and larvae ubiquitously expressing RNAi targeting 4EHP, are developmentally delayed, fail to grow and eventually die. In addition, we found that expressing RNAi that targets 4EHP specifically in the prothoracic gland disrupted ecdysone biosynthesis, causing a block of the transition from the larval to pupal stages. This phenotype can be rescued by dietary administration of ecdysone. Consistent with this, 4EHP is highly expressed in the prothoracic gland and it is required for wild type expression levels of steroidogenic enzymes. Taken together, these results uncover a novel essential function for 4EHP in regulating ecdysone biosynthesis. PMID:26721418

  4. A note on the population genetic consequences of delayed larval development in insects.

    PubMed

    de Salles, Marcos Mattoso; Otto, Paulo A

    2013-09-01

    Observations by Dobzhansky's group in the 1940s suggesting that the presence of recessive genotypes could account for lower larval developmental rates in Drosophila melanogaster were not confirmed at the time and all subsequent investigations on this subject focused on the analysis of ecological models based on competition among pre-adult individuals. However, a paper published in this journal in 1991 eventually confirmed the finding made by Dobzhansky and his co-workers. In this report, we provide a theoretical analysis of the population genetic effects of a delay in the rate of larval development produced by such a genetic mechanism. PMID:24130452

  5. Functional Genetic Screen to Identify Interneurons Governing Behaviorally Distinct Aspects of Drosophila Larval Motor Programs.

    PubMed

    Clark, Matt Q; McCumsey, Stephanie J; Lopez-Darwin, Sereno; Heckscher, Ellie S; Doe, Chris Q

    2016-01-01

    Drosophila larval crawling is an attractive system to study rhythmic motor output at the level of animal behavior. Larval crawling consists of waves of muscle contractions generating forward or reverse locomotion. In addition, larvae undergo additional behaviors, including head casts, turning, and feeding. It is likely that some neurons (e.g., motor neurons) are used in all these behaviors, but the identity (or even existence) of neurons dedicated to specific aspects of behavior is unclear. To identify neurons that regulate specific aspects of larval locomotion, we performed a genetic screen to identify neurons that, when activated, could elicit distinct motor programs. We used 165 Janelia CRM-Gal4 lines-chosen for sparse neuronal expression-to ectopically express the warmth-inducible neuronal activator TrpA1, and screened for locomotor defects. The primary screen measured forward locomotion velocity, and we identified 63 lines that had locomotion velocities significantly slower than controls following TrpA1 activation (28°). A secondary screen was performed on these lines, revealing multiple discrete behavioral phenotypes, including slow forward locomotion, excessive reverse locomotion, excessive turning, excessive feeding, immobile, rigid paralysis, and delayed paralysis. While many of the Gal4 lines had motor, sensory, or muscle expression that may account for some or all of the phenotype, some lines showed specific expression in a sparse pattern of interneurons. Our results show that distinct motor programs utilize distinct subsets of interneurons, and provide an entry point for characterizing interneurons governing different elements of the larval motor program. PMID:27172197

  6. Functional Genetic Screen to Identify Interneurons Governing Behaviorally Distinct Aspects of Drosophila Larval Motor Programs

    PubMed Central

    Clark, Matt Q.; McCumsey, Stephanie J.; Lopez-Darwin, Sereno; Heckscher, Ellie S.; Doe, Chris Q.

    2016-01-01

    Drosophila larval crawling is an attractive system to study rhythmic motor output at the level of animal behavior. Larval crawling consists of waves of muscle contractions generating forward or reverse locomotion. In addition, larvae undergo additional behaviors, including head casts, turning, and feeding. It is likely that some neurons (e.g., motor neurons) are used in all these behaviors, but the identity (or even existence) of neurons dedicated to specific aspects of behavior is unclear. To identify neurons that regulate specific aspects of larval locomotion, we performed a genetic screen to identify neurons that, when activated, could elicit distinct motor programs. We used 165 Janelia CRM-Gal4 lines—chosen for sparse neuronal expression—to ectopically express the warmth-inducible neuronal activator TrpA1, and screened for locomotor defects. The primary screen measured forward locomotion velocity, and we identified 63 lines that had locomotion velocities significantly slower than controls following TrpA1 activation (28°). A secondary screen was performed on these lines, revealing multiple discrete behavioral phenotypes, including slow forward locomotion, excessive reverse locomotion, excessive turning, excessive feeding, immobile, rigid paralysis, and delayed paralysis. While many of the Gal4 lines had motor, sensory, or muscle expression that may account for some or all of the phenotype, some lines showed specific expression in a sparse pattern of interneurons. Our results show that distinct motor programs utilize distinct subsets of interneurons, and provide an entry point for characterizing interneurons governing different elements of the larval motor program. PMID:27172197

  7. Patterns of Molecular Variation. II. Associations of Electrophoretic Mobility and Larval Substrate within Species of the DROSOPHILA MULLERI Complex

    PubMed Central

    Richardson, R. H.; Smouse, Peter E.; Richardson, Martha E.

    1977-01-01

    Electromorphic variation among populations of Drosophila mojavensis, D. arizonensis and D. longicornis was examined for seven genetic loci. The average electrophoretic mobility for a population was used as the metric. D. mojavensis and D. arizonensis use larval substrates in different parts of their geographic ranges, while D. longicornis is more narrowly restricted to different species of the cactus Opuntia in different localities. There is marked electromorphic variation among populations of either D. mojavensis or D. arizonensis, and the bulk of this variation is accounted for by differences in laval substrate. There is somewhat less variation among populations of D. longicornis, and only a moderate portion of this is accounted for by larval substrate differences. There appears to be an association between the taxonomic diversity of the larval substrates and the electromorphic diversity of the Drosophila populations utilizing those substrates. Evidence is reviewed that suggests physiological mechanisms for these possibly adaptive associations. PMID:838268

  8. Food selection in larval fruit flies: dynamics and effects on larval development

    NASA Astrophysics Data System (ADS)

    Schwarz, Sebastian; Durisko, Zachary; Dukas, Reuven

    2014-01-01

    Selecting food items and attaining a nutritionally balanced diet is an important challenge for all animals including humans. We aimed to establish fruit fly larvae ( Drosophila melanogaster) as a simple yet powerful model system for examining the mechanisms of specific hunger and diet selection. In two lab experiments with artificial diets, we found that larvae deprived of either sucrose or protein later selectively fed on a diet providing the missing nutrient. When allowed to freely move between two adjacent food patches, larvae surprisingly preferred to settle on one patch containing yeast and ignored the patch providing sucrose. Moreover, when allowed to move freely between three patches, which provided either yeast only, sucrose only or a balanced mixture of yeast and sucrose, the majority of larvae settled on the yeast-plus-sucrose patch and about one third chose to feed on the yeast only food. While protein (yeast) is essential for development, we also quantified larval success on diets with or without sucrose and show that larvae develop faster on diets containing sucrose. Our data suggest that fruit fly larvae can quickly assess major nutrients in food and seek a diet providing a missing nutrient. The larvae, however, probably prefer to quickly dig into a single food substrate for enhanced protection over achieving an optimal diet.

  9. UPRT, a suicide-gene therapy candidate in higher eukaryotes, is required for Drosophila larval growth and normal adult lifespan.

    PubMed

    Ghosh, Arpan C; Shimell, MaryJane; Leof, Emma R; Haley, Macy J; O'Connor, Michael B

    2015-01-01

    Uracil phosphoribosyltransferase (UPRT) is a pyrimidine salvage pathway enzyme that catalyzes the conversion of uracil to uridine monophosphate (UMP). The enzyme is highly conserved from prokaryotes to humans and yet phylogenetic evidence suggests that UPRT homologues from higher-eukaryotes, including Drosophila, are incapable of binding uracil. Purified human UPRT also do not show any enzymatic activity in vitro, making microbial UPRT an attractive candidate for anti-microbial drug development, suicide-gene therapy, and cell-specific mRNA labeling techniques. Nevertheless, the enzymatic site of UPRT remains conserved across the animal kingdom indicating an in vivo role for the enzyme. We find that the Drosophila UPRT homologue, krishah (kri), codes for an enzyme that is required for larval growth, pre-pupal/pupal viability and long-term adult lifespan. Our findings suggest that UPRT from all higher eukaryotes is likely enzymatically active in vivo and challenges the previous notion that the enzyme is non-essential in higher eukaryotes and cautions against targeting the enzyme for therapeutic purposes. Our findings also suggest that expression of the endogenous UPRT gene will likely cause background incorporation when using microbial UPRT as a cell-specific mRNA labeling reagent in higher eukaryotes. PMID:26271729

  10. Four Individually Identified Paired Dopamine Neurons Signal Reward in Larval Drosophila.

    PubMed

    Rohwedder, Astrid; Wenz, Nana L; Stehle, Bernhard; Huser, Annina; Yamagata, Nobuhiro; Zlatic, Marta; Truman, James W; Tanimoto, Hiromu; Saumweber, Timo; Gerber, Bertram; Thum, Andreas S

    2016-03-01

    Dopaminergic neurons serve multiple functions, including reinforcement processing during associative learning [1-12]. It is thus warranted to understand which dopaminergic neurons mediate which function. We study larval Drosophila, in which only approximately 120 of a total of 10,000 neurons are dopaminergic, as judged by the expression of tyrosine hydroxylase (TH), the rate-limiting enzyme of dopamine biosynthesis [5, 13]. Dopaminergic neurons mediating reinforcement in insect olfactory learning target the mushroom bodies, a higher-order "cortical" brain region [1-5, 11, 12, 14, 15]. We discover four previously undescribed paired neurons, the primary protocerebral anterior medial (pPAM) neurons. These neurons are TH positive and subdivide the medial lobe of the mushroom body into four distinct subunits. These pPAM neurons are acutely necessary for odor-sugar reward learning and require intact TH function in this process. However, they are dispensable for aversive learning and innate behavior toward the odors and sugars employed. Optogenetical activation of pPAM neurons is sufficient as a reward. Thus, the pPAM neurons convey a likely dopaminergic reward signal. In contrast, DL1 cluster neurons convey a corresponding punishment signal [5], suggesting a cellular division of labor to convey dopaminergic reward and punishment signals. On the level of individually identified neurons, this uncovers an organizational principle shared with adult Drosophila and mammals [1-4, 7, 9, 10] (but see [6]). The numerical simplicity and connectomic tractability of the larval nervous system [16-19] now offers a prospect for studying circuit principles of dopamine function at unprecedented resolution. PMID:26877086

  11. Lineage-associated tracts defining the anatomy of the Drosophila first instar larval brain.

    PubMed

    Hartenstein, Volker; Younossi-Hartenstein, Amelia; Lovick, Jennifer K; Kong, Angel; Omoto, Jaison J; Ngo, Kathy T; Viktorin, Gudrun

    2015-10-01

    Fixed lineages derived from unique, genetically specified neuroblasts form the anatomical building blocks of the Drosophila brain. Neurons belonging to the same lineage project their axons in a common tract, which is labeled by neuronal markers. In this paper, we present a detailed atlas of the lineage-associated tracts forming the brain of the early Drosophila larva, based on the use of global markers (anti-Neuroglian, anti-Neurotactin, inscuteable-Gal4>UAS-chRFP-Tub) and lineage-specific reporters. We describe 68 discrete fiber bundles that contain axons of one lineage or pairs/small sets of adjacent lineages. Bundles enter the neuropil at invariant locations, the lineage tract entry portals. Within the neuropil, these fiber bundles form larger fascicles that can be classified, by their main orientation, into longitudinal, transverse, and vertical (ascending/descending) fascicles. We present 3D digital models of lineage tract entry portals and neuropil fascicles, set into relationship to commonly used, easily recognizable reference structures such as the mushroom body, the antennal lobe, the optic lobe, and the Fasciclin II-positive fiber bundles that connect the brain and ventral nerve cord. Correspondences and differences between early larval tract anatomy and the previously described late larval and adult lineage patterns are highlighted. Our L1 neuro-anatomical atlas of lineages constitutes an essential step towards following morphologically defined lineages to the neuroblasts of the early embryo, which will ultimately make it possible to link the structure and connectivity of a lineage to the expression of genes in the particular neuroblast that gives rise to that lineage. Furthermore, the L1 atlas will be important for a host of ongoing work that attempts to reconstruct neuronal connectivity at the level of resolution of single neurons and their synapses. PMID:26141956

  12. Distinct visual pathways mediate Drosophila larval light avoidance and circadian clock entrainment.

    PubMed

    Keene, Alex C; Mazzoni, Esteban O; Zhen, Jamie; Younger, Meg A; Yamaguchi, Satoko; Blau, Justin; Desplan, Claude; Sprecher, Simon G

    2011-04-27

    Visual organs perceive environmental stimuli required for rapid initiation of behaviors and can also entrain the circadian clock. The larval eye of Drosophila is capable of both functions. Each eye contains only 12 photoreceptors (PRs), which can be subdivided into two subtypes. Four PRs express blue-sensitive rhodopsin5 (rh5) and eight express green-sensitive rhodopsin6 (rh6). We found that either PR-subtype is sufficient to entrain the molecular clock by light, while only the Rh5-PR subtype is essential for light avoidance. Acetylcholine released from PRs confers both functions. Both subtypes of larval PRs innervate the main circadian pacemaker neurons of the larva, the neuropeptide PDF (pigment-dispersing factor)-expressing lateral neurons (LNs), providing sensory input to control circadian rhythms. However, we show that PDF-expressing LNs are dispensable for light avoidance, and a distinct set of three clock neurons is required. Thus we have identified distinct sensory and central circuitry regulating light avoidance behavior and clock entrainment. Our findings provide insights into the coding of sensory information for distinct behavioral functions and the underlying molecular and neuronal circuitry. PMID:21525293

  13. A role for blind DN2 clock neurons in temperature entrainment of the Drosophila larval brain.

    PubMed

    Picot, Marie; Klarsfeld, André; Chélot, Elisabeth; Malpel, Sébastien; Rouyer, François

    2009-07-01

    Circadian clocks synchronize to the solar day by sensing the diurnal changes in light and temperature. In adult Drosophila, the brain clock that controls rest-activity rhythms relies on neurons showing Period oscillations. Nine of these neurons are present in each larval brain hemisphere. They can receive light inputs through Cryptochrome (CRY) and the visual system, but temperature input pathways are unknown. Here, we investigate how the larval clock network responds to light and temperature. We focused on the CRY-negative dorsal neurons (DN2s), in which light-dark (LD) cycles set molecular oscillations almost in antiphase to all other clock neurons. We first showed that the phasing of the DN2s in LD depends on the pigment-dispersing factor (PDF) neuropeptide in four lateral neurons (LNs), and on the PDF receptor in the DN2s. In the absence of PDF signaling, these cells appear blind, but still synchronize to temperature cycles. Period oscillations in the DN2s were stronger in thermocycles than in LD, but with a very similar phase. Conversely, the oscillations of LNs were weaker in thermocycles than in LD, and were phase-shifted in synchrony with the DN2s, whereas the phase of the three other clock neurons was advanced by a few hours. In the absence of any other functional clock neurons, the PDF-positive LNs were entrained by LD cycles but not by temperature cycles. Our results show that the larval clock neurons respond very differently to light and temperature, and strongly suggest that the CRY-negative DN2s play a prominent role in the temperature entrainment of the network. PMID:19571122

  14. Divergence of larval resource acquisition for water conservation and starvation resistance in Drosophila melanogaster.

    PubMed

    Parkash, Ravi; Aggarwal, Dau Dayal; Ranga, Poonam; Singh, Divya

    2012-07-01

    Laboratory selection experiments have evidenced storage of energy metabolites in adult flies of desiccation and starvation resistant strains of D. melanogaster but resource acquisition during larval stages has received lesser attention. For wild populations of D. melanogaster, it is not clear whether larvae acquire similar or different energy metabolites for desiccation and starvation resistance. We tested the hypothesis whether larval acquisition of energy metabolites is consistent with divergence of desiccation and starvation resistance in darker and lighter isofemale lines of D. melanogaster. Our results are interesting in several respects. First, we found contrasting patterns of larval resource acquisition, i.e., accumulation of higher carbohydrates during 3rd instar larval stage of darker flies versus higher levels of triglycerides in 1st and 2nd larval instars of lighter flies. Second, 3rd instar larvae of darker flies showed ~40 h longer duration of development at 21°C; and greater accumulation of carbohydrates (trehalose and glycogen) in fed larvae as compared with larvae non-fed after 150 h of egg laying. Third, darker isofemale lines have shown significant increase in total water content (18%); hemolymph (86%) and dehydration tolerance (11%) as compared to lighter isofemale lines. Loss of hemolymph water under desiccation stress until death was significantly higher in darker as compared to lighter isofemale lines but tissue water loss was similar. Fourth, for larvae of darker flies, about 65% energy content is contributed by carbohydrates for conferring greater desiccation resistance while the larvae of lighter flies acquire 2/3 energy from lipids for sustaining starvation resistance; and such energy differences persist in the newly eclosed flies. Thus, larval stages of wild-caught darker and lighter flies have evolved independent physiological processes for the accumulation of energy metabolites to cope with desiccation or starvation stress. PMID

  15. Acquisition of high-quality digital video of Drosophila larval and adult behaviors from a lateral perspective.

    PubMed

    Zenger, Beatrix; Wetzel, Sabine; Duncan, Jason

    2014-01-01

    Drosophila melanogaster is a powerful experimental model system for studying the function of the nervous system. Gene mutations that cause dysfunction of the nervous system often produce viable larvae and adults that have locomotion defective phenotypes that are difficult to adequately describe with text or completely represent with a single photographic image. Current modes of scientific publishing, however, support the submission of digital video media as supplemental material to accompany a manuscript. Here we describe a simple and widely accessible microscopy technique for acquiring high-quality digital video of both Drosophila larval and adult phenotypes from a lateral perspective. Video of larval and adult locomotion from a side-view is advantageous because it allows the observation and analysis of subtle distinctions and variations in aberrant locomotive behaviors. We have successfully used the technique to visualize and quantify aberrant crawling behaviors in third instar larvae, in addition to adult mutant phenotypes and behaviors including grooming. PMID:25350294

  16. The tamas gene, identified as a mutation that disrupts larval behavior in Drosophila melanogaster, codes for the mitochondrial DNA polymerase catalytic subunit (DNApol-gamma125).

    PubMed Central

    Iyengar, B; Roote, J; Campos, A R

    1999-01-01

    From a screen of pupal lethal lines of Drosophila melanogaster we identified a mutant strain that displayed a reproducible reduction in the larval response to light. Moreover, this mutant strain showed defects in the development of the adult visual system and failure to undergo behavioral changes characteristic of the wandering stage. The foraging third instar larvae remained in the food substrate for a prolonged period and died at or just before pupariation. Using a new assay for individual larval photobehavior we determined that the lack of response to light in these mutants was due to a primary deficit in locomotion. The mutation responsible for these phenotypes was mapped to the lethal complementation group l(2)34Dc, which we renamed tamas (translated from Sanskrit as "dark inertia"). Sequencing of mutant alleles demonstrated that tamas codes for the mitochondrial DNA polymerase catalytic subunit (DNApol-gamma125). PMID:10581287

  17. Using Drosophila Larval Imaginal Discs to Study Low-Dose Radiation-Induced Cell Cycle Arrest

    PubMed Central

    Yan, Shian-Jang; Li, Willis X.

    2012-01-01

    Under genotoxic stress, activation of cell cycle checkpoint responses leads to cell cycle arrest, which allows cells to repair DNA damage before continuing to cycle. Drosophila larval epithelial sacs, called imaginal discs, are an excellent in vivo model system for studying radiation-induced cell cycle arrest. Larval imaginal discs go into cell cycle arrest after being subjected to low-dose irradiation, are subject to easy genetic manipulation, are not crucial for survival of the organism, and can be dissected easily for further molecular or cellular analysis. In this chapter, we describe methods for assessing low-dose irradiation-induced cell cycle arrest. Mitotic cells are identified by immunofluorescence staining for the mitotic marker phosphorylated histone H3 (phospho-histone H3 or pH3). When wandering third-instar control larvae, without transgene expression, are exposed to 500 rads of X-ray or γ-ray irradiation, the number of pH3-positive cells in wing imaginal discs is reduced from hundreds before irradiation to approximately 30 after irradiation, with an equal distribution between the anterior and posterior compartments (Yan et al., 2011, FASEB J). Using the GAL4/UAS system, RNAi, cDNA, or microRNA sponge transgenes can be expressed in the posterior compartment of the wing disc using drivers such as engrailed (en)-Gal4, while the anterior compartment serves as an internal control. This approach makes it possible to do genome-wide genetic screening for molecules involved in radiation-induced cell cycle arrest. PMID:21870287

  18. Using Drosophila larval imaginal discs to study low-dose radiation-induced cell cycle arrest.

    PubMed

    Yan, Shian-Jang; Li, Willis X

    2011-01-01

    Under genotoxic stress, activation of cell cycle checkpoint responses leads to cell cycle arrest, which allows cells to repair DNA damage before continuing to cycle. Drosophila larval epithelial sacs, called imaginal discs, are an excellent in vivo model system for studying radiation-induced cell cycle arrest. Larval imaginal discs go into cell cycle arrest after being subjected to low-dose irradiation, are subject to easy genetic manipulation, are not crucial for survival of the organism, and can be dissected easily for further molecular or cellular analysis. In this chapter, we describe methods for assessing low-dose irradiation-induced cell cycle arrest. Mitotic cells are identified by immunofluorescence staining for the mitotic marker phosphorylated histone H3 (phospho-histone H3 or pH3). When wandering third-instar control larvae, without transgene expression, are exposed to 500 rads of X-ray or γ-ray irradiation, the number of pH3-positive cells in wing imaginal discs is reduced from hundreds before irradiation to approximately 30 after irradiation, with an equal distribution between the anterior and posterior compartments (Yan et al., 2011, FASEB J). Using the GAL4/UAS system, RNAi, cDNA, or microRNA sponge transgenes can be expressed in the posterior compartment of the wing disc using drivers such as engrailed (en)-Gal4, while the anterior compartment serves as an internal control. This approach makes it possible to do genome-wide genetic screening for molecules involved in radiation-induced cell cycle arrest. PMID:21870287

  19. Synaptic excitation is regulated by the postsynaptic dSK channel at the Drosophila larval NMJ

    PubMed Central

    Gertner, Daniel M.; Desai, Sunil

    2014-01-01

    In the mammalian central nervous system, the postsynaptic small-conductance Ca2+-dependent K+ (SK) channel has been shown to reduce postsynaptic depolarization and limit Ca2+ influx through N-methyl-d-aspartate receptors. To examine further the role of the postsynaptic SK channel in synaptic transmission, we studied its action at the Drosophila larval neuromuscular junction (NMJ). Repetitive synaptic stimulation produced an increase in postsynaptic membrane conductance leading to depression of excitatory postsynaptic potential amplitude and hyperpolarization of the resting membrane potential (RMP). This reduction in synaptic excitation was due to the postsynaptic Drosophila SK (dSK) channel; synaptic depression, increased membrane conductance and RMP hyperpolarization were reduced in dSK mutants or after expressing a Ca2+ buffer in the muscle. Ca2+ entering at the postsynaptic membrane was sufficient to activate dSK channels based upon studies in which the muscle membrane was voltage clamped to prevent opening voltage-dependent Ca2+ channels. Increasing external Ca2+ produced an increase in resting membrane conductance and RMP that was not seen in dSK mutants or after adding the glutamate-receptor blocker philanthotoxin. Thus it appeared that dSK channels were also activated by spontaneous transmitter release and played a role in setting membrane conductance and RMP. In mammals, dephosphorylation by protein phosphatase 2A (PP2A) increased the Ca2+ sensitivity of the SK channel; PP2A appeared to increase the sensitivity of the dSK channel since PP2A inhibitors reduced activation of the dSK channel by evoked synaptic activity or increased external Ca2+. It is proposed that spontaneous and evoked transmitter release activate the postsynaptic dSK channel to limit synaptic excitation and stabilize synapses. PMID:24671529

  20. The Drosophila larval visual system: high-resolution analysis of a simple visual neuropil.

    PubMed

    Sprecher, Simon G; Cardona, Albert; Hartenstein, Volker

    2011-10-01

    The task of the visual system is to translate light into neuronal encoded information. This translation of photons into neuronal signals is achieved by photoreceptor neurons (PRs), specialized sensory neurons, located in the eye. Upon perception of light the PRs will send a signal to target neurons, which represent a first station of visual processing. Increasing complexity of visual processing stems from the number of distinct PR subtypes and their various types of target neurons that are contacted. The visual system of the fruit fly larva represents a simple visual system (larval optic neuropil, LON) that consists of 12 PRs falling into two classes: blue-senstive PRs expressing Rhodopsin 5 (Rh5) and green-sensitive PRs expressing Rhodopsin 6 (Rh6). These afferents contact a small number of target neurons, including optic lobe pioneers (OLPs) and lateral clock neurons (LNs). We combine the use of genetic markers to label both PR subtypes and the distinct, identifiable sets of target neurons with a serial EM reconstruction to generate a high-resolution map of the larval optic neuropil. We find that the larval optic neuropil shows a clear bipartite organization consisting of one domain innervated by PRs and one devoid of PR axons. The topology of PR projections, in particular the relationship between Rh5 and Rh6 afferents, is maintained from the nerve entering the brain to the axon terminals. The target neurons can be subdivided according to neurotransmitter or neuropeptide they use as well as the location within the brain. We further track the larval optic neuropil through development from first larval instar to its location in the adult brain as the accessory medulla. PMID:21781960

  1. The Drosophila larval visual system: high-resolution analysis of a simple visual neuropil

    PubMed Central

    Sprecher, Simon G.; Cardona, Albert; Hartenstein, Volker

    2012-01-01

    The task of the visual system is to translate light into neuronal encoded information. This translation of photons into neuronal signals is achieved by photoreceptor neurons (PRs), specialized sensory neurons, located in the eye. Upon perception of light the PRs will send a signal to target neurons, which represent a first station of visual processing. Increasing complexity of visual processing stems from the number of distinct PR-subtypes and their various types of target neurons that are contacted. The visual system of the fruit fly larva represents a simple visual system (larval optic neuropil, LON) that consists of 12 PRs falling into two classes: blue-senstive PRs expressing Rhodopsin 5 (Rh5) and green-sensitive PRs expressing Rhodopsin 6 (Rh6). These afferents contact a small number of target neurons, including optic lobe pioneers (OLPs) and lateral clock neurons (LNs). We combine the use of genetic markers to label both PR-subtypes and the distinct, identifiable sets of target neurons with a serial EM reconstruction to generate a high-resolution map of the larval optic neuropil. We find that the larval optic neuropil shows a clear bipartite organization consisting of one domain innervated by PRs and one devoid of PR axons. The topology of PR projections, in particular the relationship between Rh5 and Rh6 afferents, is maintained from the nerve entering the brain to the axon terminals. The target neurons can be subdivided according to neurotransmitter or neuropeptide they use as well as the location within the brain. We further track the larval optic neuropil through development from first larval instar to its location in the adult brain as the accessory medulla. PMID:21781960

  2. Larval development of Evermannia zosterura (Perciformes: Gobiidae).

    PubMed

    González-Navarro, Enrique; Saldierna-Martínez, Ricardo Javier; Aceves-Medina, Gerardo

    2014-06-01

    Gobiidae is the most specious fish family in the world with almost 2 000 species, however only 11% of them have been described for their larval stages. The entire life cycle information is essential to understand the biology and ecology of this important fish group. Previous studies on zooplankton samples from Ensenada de La Paz, México, have shown the presence of several Gobiidae larvae and juveniles which were identified as Evermania zosterura. The main objective of this work was to describe the larval stages of this species, widely distributed in the Eastern tropical Pacific. The development of E. zosterura larvae was described based on 66 specimens. A total of 53 specimens were used to describe morphometrics and pigmentation patterns, while 13 specimens were cleared and stained, to obtain meristic characteristics. Cleared specimens had 30 to 31 total vertebrae; dorsal-fin elements: IV; 1, 13-14, anal-fin elements: 1, 13-14, and most had pterygiophore formula 4-111100. The combination of these characteristics confirmed these specimens as E. zosterura. The pigment pattern is similar throughout ontogeny. Larvae are characterized by having three to five dendritic melanophores along the post-anal ventral margin, four to nine smaller melanophores along the ventral margin between the isthmus and anus, and one on the midpoint of the dorsal margin of the tail. There is one small pigment spot on the angle of the jaw, and other on the tip of lower lip. There is an elongated internal pigment under the notochord, between the head and gas bladder. Notochord flexion starts near 3.5mm BL and ends at 4.6mm BL; transformalion to the juvenile stage is at about 13.6mm BL. Our conclusion is that the most useful characters to distinguish this species early-larval stages from those of similar species in the area, are the number of myomeres, the large melanophores (approximately uniformly in size) on the post anal ventral margin, and the elongate internal pigment under the notochord

  3. Drosophila Immunity: Analysis of Larval Hemocytes by P-Element-Mediated Enhancer Trap

    PubMed Central

    Braun, A.; Lemaitre, B.; Lanot, R.; Zachary, D.; Meister, M.

    1997-01-01

    Our aim was to identify new genes involved in the cellular aspects of defense mechanisms of Drosophila, as well as in melanotic tumor formation processes that are linked to blood cell disregulation. We have screened 1341 enhancer detector fly lines for expression of the lacZ reporter gene in larval hemocytes at the end of the third instar. We have selected 21 lines in which we observed a reproducible lacZ expression in blood cells. These lines were classified according to the subsets of hemocytes in which lacZ was expressed, and we identified five lines that can be used as lamellocyte markers. Three lines were selected for further analysis. The first exhibited strong lacZ expression in all lamellocytes. The second expressed lacZ in plasmatocytes and lamellocytes, and exhibited a melanotic tumor phenotype in larvae homozygous for the insertion. A third line showed a striking insertion-linked phenotype of melanized lymph glands (the hematopoietic organ), which resulted in the total absence of circulating hemocytes in the mutant larvae. We anticipate that this mutation, which we named domino, will prove a useful tool in the analysis of the role of hemocytes during the various aspects of immune response and melanotic tumor formation. PMID:9335599

  4. Lineage mapping identifies molecular and architectural similarities between the larval and adult Drosophila central nervous system

    PubMed Central

    Lacin, Haluk; Truman, James W

    2016-01-01

    Neurogenesis in Drosophila occurs in two phases, embryonic and post-embryonic, in which the same set of neuroblasts give rise to the distinct larval and adult nervous systems, respectively. Here, we identified the embryonic neuroblast origin of the adult neuronal lineages in the ventral nervous system via lineage-specific GAL4 lines and molecular markers. Our lineage mapping revealed that neurons born late in the embryonic phase show axonal morphology and transcription factor profiles that are similar to the neurons born post-embryonically from the same neuroblast. Moreover, we identified three thorax-specific neuroblasts not previously characterized and show that HOX genes confine them to the thoracic segments. Two of these, NB2-3 and NB3-4, generate leg motor neurons. The other neuroblast is novel and appears to have arisen recently during insect evolution. Our findings provide a comprehensive view of neurogenesis and show how proliferation of individual neuroblasts is dictated by temporal and spatial cues. DOI: http://dx.doi.org/10.7554/eLife.13399.001 PMID:26975248

  5. Gemin3 Is an Essential Gene Required for Larval Motor Function and Pupation in Drosophila

    PubMed Central

    Shpargel, Karl B.; Praveen, Kavita; Rajendra, T. K.

    2009-01-01

    The assembly of metazoan Sm-class small nuclear ribonucleoproteins (snRNPs) is an elaborate, step-wise process that takes place in multiple subcellular compartments. The initial steps, including formation of the core RNP, are mediated by the survival motor neuron (SMN) protein complex. Loss-of-function mutations in human SMN1 result in a neuromuscular disease called spinal muscular atrophy. The SMN complex is comprised of SMN and a number of tightly associated proteins, collectively called Gemins. In this report, we identify and characterize the fruitfly ortholog of the DEAD box protein, Gemin3. Drosophila Gemin3 (dGem3) colocalizes and interacts with dSMN in vitro and in vivo. RNA interference for dGem3 codepletes dSMN and inhibits efficient Sm core assembly in vitro. Transposon insertion mutations in Gemin3 are larval lethals and also codeplete dSMN. Transgenic overexpression of dGem3 rescues lethality, but overexpression of dSMN does not, indicating that loss of dSMN is not the primary cause of death. Gemin3 mutant larvae exhibit motor defects similar to previously characterized Smn alleles. Remarkably, appreciable numbers of Gemin3 mutants (along with one previously undescribed Smn allele) survive as larvae for several weeks without pupating. Our results demonstrate the conservation of Gemin3 protein function in metazoan snRNP assembly and reveal that loss of either Smn or Gemin3 can contribute to neuromuscular dysfunction. PMID:18923150

  6. A single GABAergic neuron mediates feedback of odor-evoked signals in the mushroom body of larval Drosophila

    PubMed Central

    Masuda-Nakagawa, Liria M.; Ito, Kei; Awasaki, Takeshi; O'Kane, Cahir J.

    2014-01-01

    Inhibition has a central role in defining the selectivity of the responses of higher order neurons to sensory stimuli. However, the circuit mechanisms of regulation of these responses by inhibitory neurons are still unclear. In Drosophila, the mushroom bodies (MBs) are necessary for olfactory memory, and by implication for the selectivity of learned responses to specific odors. To understand the circuitry of inhibition in the calyx (the input dendritic region) of the MBs, and its relationship with MB excitatory activity, we used the simple anatomy of the Drosophila larval olfactory system to identify any inhibitory inputs that could contribute to the selectivity of MB odor responses. We found that a single neuron accounts for all detectable GABA innervation in the calyx of the MBs, and that this neuron has pre-synaptic terminals in the calyx and post-synaptic branches in the MB lobes (output axonal area). We call this neuron the larval anterior paired lateral (APL) neuron, because of its similarity to the previously described adult APL neuron. Reconstitution of GFP partners (GRASP) suggests that the larval APL makes extensive contacts with the MB intrinsic neurons, Kenyon Cells (KCs), but few contacts with incoming projection neurons (PNs). Using calcium imaging of neuronal activity in live larvae, we show that the larval APL responds to odors, in a mannner that requires output from KCs. Our data suggest that the larval APL is the sole GABAergic neuron that innervates the MB input region and carries inhibitory feedback from the MB output region, consistent with a role in modulating the olfactory selectivity of MB neurons. PMID:24782716

  7. The conserved P body component HPat/Pat1 negatively regulates synaptic terminal growth at the larval Drosophila neuromuscular junction.

    PubMed

    Pradhan, Sarala J; Nesler, Katherine R; Rosen, Sarah F; Kato, Yasuko; Nakamura, Akira; Ramaswami, Mani; Barbee, Scott A

    2012-12-15

    The temporal and spatial regulation of protein synthesis plays an important role in the control of neural physiology. In axons and dendrites, translationally repressed mRNAs are actively transported to their destinations in a variety of ribonucleoprotein particles (RNPs). A subset of these neuronal RNPs has been shown to contain proteins associated with mRNA processing bodies (P bodies). P bodies are a class of highly conserved cytoplasmic granules that have been linked to both mRNA decay and translational repression via general and miRNA-mediated pathways. Here, we characterize functions for HPat/Pat1 (also known as Patr-1), a core component of P bodies, at the glutamatergic larval Drosophila neuromuscular junction (NMJ). We show that hpat mutants exhibit a strong synaptic hyperplasia at the NMJ. The synaptic defects observed in hpat mutants are associated with rearrangement of the axonal microtubule cytoskeleton suggesting that HPat negatively regulates presynaptic microtubule-based growth during NMJ development. Consistent with this, overexpression of HPat also blocks the rapid growth of presynaptic boutons induced by spaced depolarization. Finally, we demonstrate that HPat interacts genetically with the catalytic subunit of the deadenylase complex (twin/CCR4) and the miRNA pathway (Argonaute 1) to control bouton formation. We propose that HPat is required to target mRNAs involved in the control of microtubule architecture and synaptic terminal growth for repression, presumably in P bodies, via both general and miRNA-mediated mechanisms. PMID:23097047

  8. The conserved P body component HPat/Pat1 negatively regulates synaptic terminal growth at the larval Drosophila neuromuscular junction

    PubMed Central

    Pradhan, Sarala J.; Nesler, Katherine R.; Rosen, Sarah F.; Kato, Yasuko; Nakamura, Akira; Ramaswami, Mani; Barbee, Scott A.

    2012-01-01

    Summary The temporal and spatial regulation of protein synthesis plays an important role in the control of neural physiology. In axons and dendrites, translationally repressed mRNAs are actively transported to their destinations in a variety of ribonucleoprotein particles (RNPs). A subset of these neuronal RNPs has been shown to contain proteins associated with mRNA processing bodies (P bodies). P bodies are a class of highly conserved cytoplasmic granules that have been linked to both mRNA decay and translational repression via general and miRNA-mediated pathways. Here, we characterize functions for HPat/Pat1 (also known as Patr-1), a core component of P bodies, at the glutamatergic larval Drosophila neuromuscular junction (NMJ). We show that hpat mutants exhibit a strong synaptic hyperplasia at the NMJ. The synaptic defects observed in hpat mutants are associated with rearrangement of the axonal microtubule cytoskeleton suggesting that HPat negatively regulates presynaptic microtubule-based growth during NMJ development. Consistent with this, overexpression of HPat also blocks the rapid growth of presynaptic boutons induced by spaced depolarization. Finally, we demonstrate that HPat interacts genetically with the catalytic subunit of the deadenylase complex (twin/CCR4) and the miRNA pathway (Argonaute 1) to control bouton formation. We propose that HPat is required to target mRNAs involved in the control of microtubule architecture and synaptic terminal growth for repression, presumably in P bodies, via both general and miRNA-mediated mechanisms. PMID:23097047

  9. Lamellipodia-based migrations of larval epithelial cells are required for normal closure of the adult epidermis of Drosophila

    PubMed Central

    Bischoff, Marcus

    2012-01-01

    Cell migrations are an important feature of animal development. They are, furthermore, essential to wound healing and tumour progression. Despite recent progress, it is still mysterious how cell migration is spatially and temporally regulated during morphogenesis and how cell migration is coordinated with other cellular behaviours to shape tissues and organs. The formation of the abdominal epithelium of Drosophila during metamorphosis provides an attractive system to study morphogenesis. Here, the diploid adult histoblasts replace the polyploid larval epithelial cells (LECs). Using in vivo 4D microscopy, I show that, besides apical constriction and apoptosis, the LECs undergo extensive coordinated migrations. The migrations follow a transition from a stationary (epithelial) to a migratory mode. The migratory behaviour is stimulated by autocrine Dpp signalling. Directed apical lamellipodia-like protrusions propel the cells. Initially, planar cell polarity determines the orientation of LEC migration. While LECs are migrating they also constrict apically, and changes in activity of the small GTPase Rho1 can favour one behaviour over the other. This study shows that the LECs play a more active role in morphogenesis than previously thought, with their migrations contributing to abdominal closure. It furthermore provides insights into how the migratory behaviour of cells is regulated during morphogenesis. PMID:22230614

  10. Expression and Function of Scalloped During Drosophila Development

    PubMed Central

    Guss, Kirsten A.; Benson, Michael; Gubitosi, Nicholas; Brondell, Karrie; Broadie, Kendal; Skeath, James B.

    2013-01-01

    Background The scalloped (sd) and vestigial (vg) genes function together in Drosophila wing development. Little is known about sd protein (SD) expression during development, or whether sd and vg interact in other developing tissues. To begin to address these questions, we generated an anti-SD antibody. Results During embryogenesis, SD is expressed in both central and peripheral nervous systems, and the musculature. SD is also expressed in developing flight appendages. Despite SD expression herein, the peripheral nervous system, musculature, and dorsal limb primordia appeared generally normal in the absence of sd function. SD is also expressed in subsets of ventral nerve cord cells, including neuroblast 1–2 descendants and ventral unpaired median motor neurons (mVUMs). While sd function is not required to specify these neurons, it is necessary for the correct innervation of somatic muscles by the mVUMs. We also show that SD and VG are co-expressed in overlapping and distinctive subsets of cells in embryonic and larval tissues. Conclusions We describe the full breadth of SD expression during Drosophila embryogenesis, and identify a requirement for sd function in a subset of motor neurons. This work provides the necessary foundation for functional studies regarding the roles of sd during Drosophila development. PMID:23389965

  11. 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.

  12. Searching for motifs in the behaviour of larval Drosophila melanogaster and Caenorhabditis elegans reveals continuity between behavioural states

    PubMed Central

    Szigeti, Balázs; Webb, Barbara

    2015-01-01

    We present a novel method for the unsupervised discovery of behavioural motifs in larval Drosophila melanogaster and Caenorhabditis elegans. A motif is defined as a particular sequence of postures that recurs frequently. The animal's changing posture is represented by an eigenshape time series, and we look for motifs in this time series. To find motifs, the eigenshape time series is segmented, and the segments clustered using spline regression. Unlike previous approaches, our method can classify sequences of unequal duration as the same motif. The behavioural motifs are used as the basis of a probabilistic behavioural annotator, the eigenshape annotator (ESA). Probabilistic annotation avoids rigid threshold values and allows classification uncertainty to be quantified. We apply eigenshape annotation to both larval Drosophila and C. elegans and produce a good match to hand annotation of behavioural states. However, we find many behavioural events cannot be unambiguously classified. By comparing the results with ESA of an artificial agent's behaviour, we argue that the ambiguity is due to greater continuity between behavioural states than is generally assumed for these organisms. PMID:26609067

  13. 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

  14. TIF-IA-Dependent Regulation of Ribosome Synthesis in Drosophila Muscle Is Required to Maintain Systemic Insulin Signaling and Larval Growth

    PubMed Central

    Ghosh, Abhishek; Rideout, Elizabeth J.; Grewal, Savraj S.

    2014-01-01

    The conserved TOR kinase signaling network links nutrient availability to cell, tissue and body growth in animals. One important growth-regulatory target of TOR signaling is ribosome biogenesis. Studies in yeast and mammalian cell culture have described how TOR controls rRNA synthesis—a limiting step in ribosome biogenesis—via the RNA Polymerase I transcription factor TIF-IA. However, the contribution of TOR-dependent ribosome synthesis to tissue and body growth in animals is less clear. Here we show in Drosophila larvae that ribosome synthesis in muscle is required non-autonomously to maintain normal body growth and development. We find that amino acid starvation and TOR inhibition lead to reduced levels of TIF-IA, and decreased rRNA synthesis in larval muscle. When we mimic this decrease in muscle ribosome synthesis using RNAi-mediated knockdown of TIF-IA, we observe delayed larval development and reduced body growth. This reduction in growth is caused by lowered systemic insulin signaling via two endocrine responses: reduced expression of Drosophila insulin-like peptides (dILPs) from the brain and increased expression of Imp-L2—a secreted factor that binds and inhibits dILP activity—from muscle. We also observed that maintaining TIF-IA levels in muscle could partially reverse the starvation-mediated suppression of systemic insulin signaling. Finally, we show that activation of TOR specifically in muscle can increase overall body size and this effect requires TIF-IA function. These data suggest that muscle ribosome synthesis functions as a nutrient-dependent checkpoint for overall body growth: in nutrient rich conditions, TOR is required to maintain levels of TIF-IA and ribosome synthesis to promote high levels of systemic insulin, but under conditions of starvation stress, reduced muscle ribosome synthesis triggers an endocrine response that limits systemic insulin signaling to restrict growth and maintain homeostasis. PMID:25356674

  15. 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

  16. Differences in larval nutritional requirements and female oviposition preference reflect the order of fruit colonization of Zaprionus indianus and Drosophila simulans.

    PubMed

    Matavelli, Cristiane; Carvalho, Maria João A; Martins, Nelson E; Mirth, Christen K

    2015-11-01

    Species coexist using the same nutritional resource by partitioning it either in space or time, but few studies explore how species-specific nutritional requirements allow partitioning. Zaprionus indianus and Drosophila simulans co-exist in figs by invading the fruit at different stages; Z. indianus colonizes ripe figs, whereas D. simulans oviposits in decaying fruit. Larvae feed on yeast growing on the fruit, which serves as their primary protein source. Because yeast populations increase as fruit decays, we find that ripe fruit has lower protein content than rotting fruit. Therefore, we hypothesized that Z. indianus and D. simulans larvae differ in their dietary requirements for protein. We used nutritional geometry to assess the effects of protein and carbohydrate concentration in the larval diet on life history characters in both species. Survival, development time, and ovariole number respond differently to the composition of the larval diet, with Z. indianus generally performing better across a wider range of protein concentrations. Correspondingly, we found that Z. indianus females preferred to lay eggs on low protein foods, while D. simulans females chose higher protein foods for oviposition when competing with Z. indianus. We propose the different nutritional requirements and oviposition preference of these two species allows them to temporally partition their habitat. PMID:26358399

  17. Nerve-muscle interactions during flight muscle development in Drosophila

    NASA Technical Reports Server (NTRS)

    Fernandes, J. J.; Keshishian, H.

    1998-01-01

    During Drosophila pupal metamorphosis, the motoneurons and muscles differentiate synchronously, providing an opportunity for extensive intercellular regulation during synapse formation. We examined the existence of such interactions by developmentally delaying or permanently eliminating synaptic partners during the formation of indirect flight muscles. When we experimentally delayed muscle development, we found that although adult-specific primary motoneuron branching still occurred, the higher order (synaptic) branching was suspended until the delayed muscle fibers reached a favourable developmental state. In reciprocal experiments we found that denervation caused a decrease in the myoblast pool. Furthermore, the formation of certain muscle fibers (dorsoventral muscles) was specifically blocked. Exceptions were the adult muscles that use larval muscle fibers as myoblast fusion targets (dorsal longitudinal muscles). However, when these muscles were experimentally compelled to develop without their larval precursors, they showed an absolute dependence on the motoneurons for their formation. These data show that the size of the myoblast pool and early events in fiber formation depend on the presence of the nerve, and that, conversely, peripheral arbor development and synaptogenesis is closely synchronized with the developmental state of the muscle.

  18. Phormidium animalis (Cyanobacteria: Oscillatoriaceae) supports larval development of Anopheles albimanus.

    PubMed

    Vázquez-Martínez, M Guadalupe; Rodríguez, Mario H; Arredondo-Jiménez, Juan I; Méndez-Sánchez, José D

    2003-06-01

    The capability of Phormidium animalis, a cyanobacterium commonly found in larval habitats of Anopheles albimanus in southern Mexico, to support larval development of this mosquito was investigated. First-stage larvae were reared under insectary conditions with P. animalis ad libitum and their development was compared with larvae fed with wheat germ. The time of pupation and adult mosquito size, assessed by wing length, were similar in both groups, but fewer adult mosquitoes were obtained from larvae fed with the cyanobacteria. Nevertheless, these observations indicate that P. animalis is ingested and assimilated by larval An. albimanus, making this cyanobacterium a good candidate for genetic engineering for the introduction of mosquitocidal toxins for malaria control in the region. PMID:12825668

  19. Quantitative trait loci for life span in Drosophila melanogaster: interactions with genetic background and larval density.

    PubMed Central

    Leips, J; Mackay, T F

    2000-01-01

    The genetic architecture of variation in adult life span was examined for a population of recombinant inbred lines, each of which had been crossed to both inbred parental strains from which the lines were derived, after emergence from both high and low larval density. QTL affecting life span were mapped within each sex and larval density treatment by linkage to highly polymorphic roo-transposable element markers, using a composite interval mapping method. We detected a total of six QTL affecting life span; the additive effects and degrees of dominance for all were highly sex- and larval environment-specific. There were significant epistatic interactions between five of the life span QTL, the effects of which also differed according to genetic background, sex, and larval density. Five additional QTL were identified that contributed to differences among lines in their sensitivity to variation in larval density. Further fine-scale mapping is necessary to determine whether candidate genes within the regions to which the QTL map are actually responsible for the observed variation in life span. PMID:10924473

  20. An Allele of Sequoia Dominantly Enhances a Trio Mutant Phenotype to Influence Drosophila Larval Behavior

    PubMed Central

    Liebl, Eric C.

    2013-01-01

    The transition of Drosophila third instar larvae from feeding, photo-phobic foragers to non-feeding, photo-neutral wanderers is a classic behavioral switch that precedes pupariation. The neuronal network responsible for this behavior has recently begun to be defined. Previous genetic analyses have identified signaling components for food and light sensory inputs and neuropeptide hormonal outputs as being critical for the forager to wanderer transition. Trio is a Rho-Guanine Nucleotide Exchange Factor integrated into a variety of signaling networks including those governing axon pathfinding in early development. Sequoia is a pan-neuronally expressed zinc-finger transcription factor that governs dendrite and axon outgrowth. Using pre-pupal lethality as an endpoint, we have screened for dominant second-site enhancers of a weakly lethal trio mutant background. In these screens, an allele of sequoia has been identified. While these mutants have no obvious disruption of embryonic central nervous system architecture and survive to third instar larvae similar to controls, they retain forager behavior and thus fail to pupariate at high frequency. PMID:24376789

  1. Planar cell polarity: the Dachsous/Fat system contributes differently to the embryonic and larval stages of Drosophila

    PubMed Central

    Saavedra, Pedro; Brittle, Amy; Palacios, Isabel M.; Strutt, David; Casal, José; Lawrence, Peter A.

    2016-01-01

    ABSTRACT The epidermal patterns of all three larval instars (L1–L3) of Drosophila are made by one unchanging set of cells. The seven rows of cuticular denticles of all larval stages are consistently planar polarised, some pointing forwards, others backwards. In L1 all the predenticles originate at the back of the cells but, in L2 and L3, they form at the front or the back of the cell depending on the polarity of the forthcoming denticles. We find that, to polarise all rows, the Dachsous/Fat system is differentially utilised; in L1 it is active in the placement of the actin-based predenticles but is not crucial for the final orientation of the cuticular denticles, in L2 and L3 it is needed for placement and polarity. We find Four-jointed to be strongly expressed in the tendon cells and show how this might explain the orientation of all seven rows. Unexpectedly, we find that L3 that lack Dachsous differ from larvae lacking Fat and we present evidence that this is due to differently mislocalised Dachs. We make some progress in understanding how Dachs contributes to phenotypes of wildtype and mutant larvae and adults. PMID:26935392

  2. Generalization and discrimination tasks yield concordant measures of perceived distance between odours and their binary mixtures in larval Drosophila

    PubMed Central

    Chen, Yi-chun; Gerber, Bertram

    2014-01-01

    Similarity between odours is notoriously difficult to measure. Widely used behavioural approaches in insect olfaction research are cross-adaptation, masking, as well as associative tasks based on olfactory learning and the subsequent testing for how specific the established memory is. A concern with such memory-based approaches is that the learning process required to establish an odour memory may alter the way the odour is processed, such that measures of perception taken at the test are distorted. The present study was therefore designed to see whether behavioural judgements of perceptual distance are different for two different memory-based tasks, namely generalization and discrimination. We used odour–reward learning in larval Drosophila as a study case. In order to challenge the larvae's olfactory system, we chose to work with binary mixtures and their elements (1-octanol, n-amyl acetate, 3-octanol, benzaldehyde and hexyl acetate). We determined the perceptual distance between each mixture and its elements, first in a generalization task, and then in a discrimination task. It turns out that scores of perceptual distance are correlated between both tasks. A re-analysis of published studies looking at element-to-element perceptual distances in larval reward learning and in adult punishment learning confirms this result. We therefore suggest that across a given set of olfactory stimuli, associative training does not grossly alter the pattern of perceptual distances. PMID:24920835

  3. 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

  4. LARVAL DEVELOPMENT OF THE SPIDER CRAB, 'LIBINIA EMERGINATA' (MAJIDAE)

    EPA Science Inventory

    Larval development of the spider crab, Libinia emarginata, consists of two zoeal stages and megalopa. Laboratory-reared larvae (South Carolina and Rhode Island) are described and compared with planktonic larvae from Narragansett Bay, RI. No significant variations in morphology we...

  5. Transcriptional Signatures in Response to Wheat Germ Agglutinin and Starvation in Drosophila melanogaster Larval Midgut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One function of plant lectins such as wheat germ agglutinin (WGA) is to serve as defenses against herbivorous insects. The midgut is one critical site affected by dietary lectins. We observed marked cellular, structural, and gene expression changes in the midguts of Drosophila melanogaster third-i...

  6. A Behavior-Based Circuit Model of How Outcome Expectations Organize Learned Behavior in Larval "Drosophila"

    ERIC Educational Resources Information Center

    Schleyer, Michael; Saumweber, Timo; Nahrendorf, Wiebke; Fischer, Benjamin; von Alpen, Desiree; Pauls, Dennis; Thum, Andreas; Gerber, Bertram

    2011-01-01

    Drosophila larvae combine a numerically simple brain, a correspondingly moderate behavioral complexity, and the availability of a rich toolbox for transgenic manipulation. This makes them attractive as a study case when trying to achieve a circuit-level understanding of behavior organization. From a series of behavioral experiments, we suggest a…

  7. 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)…

  8. Chemical mutagenesis testing in Drosophila. VII. Results of 22 coded compounds tested in larval feeding experiments

    SciTech Connect

    Zimmering, S.; Mason, J.M.; Valencia, R. )

    1989-01-01

    Twenty-two chemicals were tested for mutagenicity in the sex-linked recessive lethal (SLRL) mutation assay after being fed to Drosophila melanogaster larvae. One compound, maleic hydrazide, was found to be mutagenic. It was tested for the ability to produce reciprocal translocations (RTs) and was positive in that assay as well.

  9. The Drosophila BTB Domain Protein Jim Lovell Has Roles in Multiple Larval and Adult Behaviors

    PubMed Central

    Bjorum, Sonia M.; Simonette, Rebecca A.; Alanis, Raul; Wang, Jennifer E.; Lewis, Benjamin M.; Trejo, Michael H.; Hanson, Keith A.; Beckingham, Kathleen M.

    2013-01-01

    Innate behaviors have their origins in the specification of neural fates during development. Within Drosophila, BTB (Bric-a-brac,Tramtrack, Broad) domain proteins such as Fruitless are known to play key roles in the neural differentiation underlying such responses. We previously identified a gene, which we have termed jim lovell (lov), encoding a BTB protein with a role in gravity responses. To understand more fully the behavioral roles of this gene we have investigated its function through several approaches. Transcript and protein expression patterns have been examined and behavioral phenotypes of new lov mutations have been characterized. Lov is a nuclear protein, suggesting a role as a transcriptional regulator, as for other BTB proteins. In late embryogenesis, Lov is expressed in many CNS and PNS neurons. An examination of the PNS expression indicates that lov functions in the late specification of several classes of sensory neurons. In particular, only two of the five abdominal lateral chordotonal neurons express Lov, predicting functional variation within this highly similar group. Surprisingly, Lov is also expressed very early in embryogenesis in ways that suggests roles in morphogenetic movements, amnioserosa function and head neurogenesis. The phenotypes of two new lov mutations that delete adjacent non-coding DNA regions are strikingly different suggesting removal of different regulatory elements. In lov47, Lov expression is lost in many embryonic neurons including the two lateral chordotonal neurons. lov47 mutant larvae show feeding and locomotor defects including spontaneous backward movement. Adult lov47 males perform aberrant courtship behavior distinguished by courtship displays that are not directed at the female. lov47 adults also show more defective negative gravitaxis than the previously isolated lov91Y mutant. In contrast, lov66 produces largely normal behavior but severe female sterility associated with ectopic lov expression in the ovary. We

  10. Embryogenesis, hatching and larval development of Artemia during orbital spaceflight

    NASA Technical Reports Server (NTRS)

    Spooner, B. S.; Debell, L.; Armbrust, L.; Guikema, J. A.; Metcalf, J.; Paulsen, A.

    1994-01-01

    Developmental biology studies, using gastrula-arrested cysts of the brine shrimp Artemia franciscana, were conducted during two flights of the space shuttle Atlantis (missions STS-37 and STS-43) in 1991. Dehydrated cysts were activated, on orbit, by addition of salt water to the cysts, and then development was terminated by the addition of fixative. Development took place in 5 ml syringes, connected by tubing to activation syringes, containing salt water, and termination syringes, containing fixative. Comparison of space results with simultaneous ground control experiments showed that equivalent percentages of naupliar larvae hatched in the syringes (40%). Thus, reactivation of development, completion of embryogenesis, emergence and hatching took place, during spaceflight, without recognizable alteration in numbers of larvae produced. Post-hatching larval development was studied in experiments where development was terminated, by introduction of fixative, 2 days, 4 days, and 8 days after reinitiation of development. During spaceflight, successive larval instars or stages, interrupted by molts, occurred, generating brine shrimp at appropriate larval instars. Naupliar larvae possessed the single naupliar eye, and development of the lateral pair of adult eyes also took place in space. Transmission electron microscopy revealed extensive differentiation, including skeletal muscle and gut endoderm, as well as the eye tissues. These studies demonstrate the potential value of Artemia for developmental biology studies during spa ceflight, and show that extensive degrees of development can take place in this microgravity environment.

  11. Embryogenesis, hatching and larval development of Artemia during orbital spaceflight

    NASA Astrophysics Data System (ADS)

    Spooner, B. S.; Debell, L.; Armbrust, L.; Guikema, J. A.; Metcalf, J.; Paulsen, A.

    1994-08-01

    Developmental biology studies, using gastrula-arrested cysts of the brine shrimp Artemia franciscana, were conducted during two flights of the space shuttle Atlantis (missions STS-37 and STS-43) in 1991. Dehydrated cysts were activated, on orbit, by addition of salt water to the cysts, and then development was terminated by the addition of fixative. Development took place in 5 ml syringes, connected by tubing to activation syringes, containing salt water, and termination syringes, containing fixative. Comparison of space results with simultaneous ground control experiments showed that equivalent percentages of naupliar larvae hatched in the syringes (40%). Thus, reactivation of development, completion of embryogenesis, emergence and hatching took place, during spaceflight, without recognizable alteration in numbers of larvae produced. Post-hatching larval development was studied in experiments where development was terminated, by intrduction of fixative, 2 days, 4 days, and 8 days after reinitiation of development. During spaceflight, successive larval instars or stages, interrupted by molts, occurred, generating brine shrimp at appropriate larval instars. Naupliar larvae possessed the single naupliar eye, and development of the lateral pair of adult eyes also took place in space. Transmission electron microscopy revealed extensive differentiation, including skeletal muscle and gut endoderm, as well as the eye tissues. These studies demonstrate the potential value of Artemia for developmental biology studies during spaceflight, and show that extensive degress of development can take place in this microgravity environment.

  12. Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS

    PubMed Central

    Fore, Taylor R.; Ojwang, Audrey A.; Warner, Margaret L.; Peng, Xinyun; Bohm, Rudolf A.; Welch, William P.; Goodnight, Lindsey K.; Bao, Hong; Zhang, Bing

    2011-01-01

    The Gal4/ UAS binary method is powerful for gene and neural circuitry manipulation in Drosophila. For most neurobiological studies, however, Gal4 expression is rarely tissue-specific enough to allow for precise correlation of the circuit with behavioral readouts. To overcome this major hurdle, we recently developed the FINGR method to achieve a more restrictive Gal4 expression in the tissue of interest. The FINGR method has three components: 1) the traditional Gal4/UAS system; 2) a set of FLP/FRT-mediated Gal80 converting tools; and 3) enhancer-trap FLP (ET-FLP). Gal4 is used to define the primary neural circuitry of interest. Paring the Gal4 with a UAS-effector, such as UAS-MJD78Q or UAS-Shits, regulates the neuronal activity, which is in turn manifested by alterations in the fly behavior. With an additional UAS-reporter such as UAS-GFP, the neural circuit involved in the specific behavior can be simultaneously mapped for morphological analysis. For Gal4 lines with broad expression, Gal4 expression can be restricted by using two complementary Gal80-converting tools: tubP>Gal80> ('flip out') and tubP>stop>Gal80 ('flip in'). Finally, investigators can turn Gal80 on or off, respectively, with the help of tissue-specific ET-FLP. In the flip-in mode, Gal80 will repress Gal4 expression wherever Gal4 and ET-FLP intersect. In the flip-out mode, Gal80 will relieve Gal4 repression in cells in which Gal4 and FLP overlap. Both approaches enable the restriction of the number of cells in the Gal4-defined circuitry, but in an inverse pattern. The FINGR method is compatible with the vast collection of Gal4 lines in the fly community and highly versatile for traditional clonal analysis and for neural circuit mapping. In this protocol, we demonstrate the mapping of FLP expression patterns in select ET-FLPx2 lines and the effectiveness of the FINGR method in photoreceptor cells. The principle of the FINGR method should also be applicable to other genetic model organisms in which Gal

  13. Vertebrate eye development as modeled in Drosophila.

    PubMed

    Wawersik, S; Maas, R L

    2000-04-12

    Pax6, a member of the paired-box family of transcription factors, is critical for oculogenesis in both vertebrates and insects. Identification of potential vertebrate Pax6 targets has been guided by studies in Drosophila, where the Pax6 homologs eyeless ( ey ) and twin of eyeless ( toy ) function within a network of genes that synergistically pattern the developing fly eye. These targets, which share homology with the fly genes sine oculis, eyes absent and dachshund, exist in mice and humans as the Six, Eya and Dach gene families. Members of these gene families are present in the developing vertebrate eye, and preliminary studies suggest that they may function in a network analogous to that in the fly. Thus, despite radically different architecture, a similar molecular scaffold underlies both vertebrate and fly eye patterning, suggesting that the considerable power of Drosophila genetics can be harnessed to study mammalian ocular development. PMID:10767315

  14. Social coercion of larval development in an ant species

    NASA Astrophysics Data System (ADS)

    Villalta, Irene; Amor, Fernando; Cerdá, Xim; Boulay, Raphaël

    2016-04-01

    Ants provide one of the best examples of the division of labor in animal societies. While the queens reproduce, workers generally refrain from laying eggs and dedicate themselves exclusively to domestic tasks. In many species, the small diploid larvae are bipotent and can develop either into workers or queens depending mostly on environmental cues. This generates a conflicting situation between the adults that tend to rear a majority of larvae into workers and the larvae whose individual interest may be to develop into reproductive queens. We tested the social regulation of larval caste fate in the fission-performing ant Aphaenogaster senilis. We first observed interactions between resident workers and queen- and worker-destined larvae in presence/absence of the queen. The results show that workers tend to specifically eliminate queen-destined larvae when the queen is present but not when she is absent or imprisoned in a small cage allowing for volatile pheromone exchanges. In addition, we found that the presence of already developed queen-destined larvae does not inhibit the development of younger still bipotent larvae into queens. Finally, we analyzed the cuticular hydrocarbon profiles of queen- and worker-destined larvae and found no significant quantitative or qualitative difference. Interestingly, the total amount of hydrocarbons on both larval castes is extremely low, which lends credence on the chemical insignificance hypothesis of larval ants. Overall, our results suggest that workers control larval development and police larvae that would develop into queens instead of workers. Such policing behavior is similar in many aspects to what is known of worker policing among adults.

  15. Social coercion of larval development in an ant species.

    PubMed

    Villalta, Irene; Amor, Fernando; Cerdá, Xim; Boulay, Raphaël

    2016-04-01

    Ants provide one of the best examples of the division of labor in animal societies. While the queens reproduce, workers generally refrain from laying eggs and dedicate themselves exclusively to domestic tasks. In many species, the small diploid larvae are bipotent and can develop either into workers or queens depending mostly on environmental cues. This generates a conflicting situation between the adults that tend to rear a majority of larvae into workers and the larvae whose individual interest may be to develop into reproductive queens. We tested the social regulation of larval caste fate in the fission-performing ant Aphaenogaster senilis. We first observed interactions between resident workers and queen- and worker-destined larvae in presence/absence of the queen. The results show that workers tend to specifically eliminate queen-destined larvae when the queen is present but not when she is absent or imprisoned in a small cage allowing for volatile pheromone exchanges. In addition, we found that the presence of already developed queen-destined larvae does not inhibit the development of younger still bipotent larvae into queens. Finally, we analyzed the cuticular hydrocarbon profiles of queen- and worker-destined larvae and found no significant quantitative or qualitative difference. Interestingly, the total amount of hydrocarbons on both larval castes is extremely low, which lends credence on the chemical insignificance hypothesis of larval ants. Overall, our results suggest that workers control larval development and police larvae that would develop into queens instead of workers. Such policing behavior is similar in many aspects to what is known of worker policing among adults. PMID:26874941

  16. Hydroxyurea-mediated neuroblast ablation establishes birth dates of secondary lineages and addresses neuronal interactions in the developing Drosophila brain.

    PubMed

    Lovick, Jennifer K; Hartenstein, Volker

    2015-06-01

    The Drosophila brain is comprised of neurons formed by approximately 100 lineages, each of which is derived from a stereotyped, asymmetrically dividing neuroblast. Lineages serve as structural and developmental units of Drosophila brain anatomy and reconstruction of lineage projection patterns represents a suitable map of Drosophila brain circuitry at the level of neuron populations ("macro-circuitry"). Two phases of neuroblast proliferation, the first in the embryo and the second during the larval phase (following a period of mitotic quiescence), produce primary and secondary lineages, respectively. Using temporally controlled pulses of hydroxyurea (HU) to ablate neuroblasts and their corresponding secondary lineages during the larval phase, we analyzed the effect on development of primary and secondary lineages in the late larval and adult brain. Our findings indicate that timing of neuroblast re-activation is highly stereotyped, allowing us to establish "birth dates" for all secondary lineages. Furthermore, our results demonstrate that, whereas the trajectory and projection pattern of primary and secondary lineages is established in a largely independent manner, the final branching pattern of secondary neurons is dependent upon the presence of appropriate neuronal targets. Taken together, our data provide new insights into the degree of neuronal plasticity during Drosophila brain development. PMID:25773365

  17. Hydroxyurea-mediated neuroblast ablation establishes birthdates of secondary lineages and addresses neuronal interactions in the developing Drosophila brain

    PubMed Central

    Lovick, Jennifer K.; Hartenstein, Volker

    2015-01-01

    The Drosophila brain is comprised of neurons formed by approximately 100 lineages, each of which is derived from a stereotyped, asymmetrically dividing neuroblast. Lineages serve as structural and developmental units of Drosophila brain anatomy and reconstruction of lineage projection patterns represents a suitable map of Drosophila brain circuitry at the level of neuron populations (“macro-circuitry”). Two phases of neuroblast proliferation, the first in the embryo and the second during the larval phase (following a period of mitotic quiescence), produce primary and secondary lineages, respectively. Using temporally controlled pulses of hydroxyurea (HU) to ablate neuroblasts and their corresponding secondary lineages during the larval phase, we analyzed the effect on development of primary and secondary lineages in the late larval and adult brain. Our findings indicate that timing of neuroblast re-activation is highly stereotyped, allowing us to establish “birth dates” for all secondary lineages. Furthermore, our results demonstrate that, whereas the trajectory and projection pattern of primary and secondary lineages is established in a largely independent manner, the final branching pattern of secondary neurons is dependent upon the presence of appropriate neuronal targets. Taken together, our data provide new insights into the degree of neuronal plasticity during Drosophila brain development. PMID:25773365

  18. A development-based compartmentalization of the Drosophila central brain

    PubMed Central

    Pereanu, Wayne; Kumar, Abilasha; Jennett, Arnim; Reichert, Heinrich; Hartenstein, Volker

    2010-01-01

    The neuropile of the Drosophila brain is subdivided into anatomically discrete compartments. Compartments are rich in terminal neurite branching and synapses; they are the neuropile domains in which signal processing takes place. Compartment boundaries are defined by more or less dense layers of glial cells, as well as long neurite fascicles. These fascicles are formed during the larval period when the approximately 100 neuronal lineages that constitute the Drosophila central brain differentiate. Each lineage forms an axon tract with a characteristic trajectory in the neuropile; groups of spatially related tracts congregate into the brain fascicles that can be followed from the larva throughout metamorphosis into the adult stage. In this paper we provide a map of the adult brain compartments and the relevant fascicles defining compartmental boundaries. We have identified the neuronal lineages contributing to each fascicle, which allowed us to directly compare compartments of the larval and adult brain. Most adult compartments can be recognized already in the early larval brain where they form a “protomap” of the later adult compartments. Our analysis highlights the morphogenetic changes shaping the Drosophila brain; the data will be important for studies that link early acting genetic mechanisms to the adult neuronal structures and circuits controlled by these mechanisms. PMID:20533357

  19. Development-based compartmentalization of the Drosophila central brain.

    PubMed

    Pereanu, Wayne; Kumar, Abilasha; Jennett, Arnim; Reichert, Heinrich; Hartenstein, Volker

    2010-08-01

    The neuropile of the Drosophila brain is subdivided into anatomically discrete compartments. Compartments are rich in terminal neurite branching and synapses; they are the neuropile domains in which signal processing takes place. Compartment boundaries are defined by more or less dense layers of glial cells as well as long neurite fascicles. These fascicles are formed during the larval period, when the approximately 100 neuronal lineages that constitute the Drosophila central brain differentiate. Each lineage forms an axon tract with a characteristic trajectory in the neuropile; groups of spatially related tracts congregate into the brain fascicles that can be followed from the larva throughout metamorphosis into the adult stage. Here we provide a map of the adult brain compartments and the relevant fascicles defining compartmental boundaries. We have identified the neuronal lineages contributing to each fascicle, which allowed us to compare compartments of the larval and adult brain directly. Most adult compartments can be recognized already in the early larval brain, where they form a "protomap" of the later adult compartments. Our analysis highlights the morphogenetic changes shaping the Drosophila brain; the data will be important for studies that link early-acting genetic mechanisms to the adult neuronal structures and circuits controlled by these mechanisms. PMID:20533357

  20. Drosophila Homolog of Human KIF22 at the Autism-Linked 16p11.2 Loci Influences Synaptic Connectivity at Larval Neuromuscular Junctions.

    PubMed

    Park, Sang Mee; Littleton, J Troy; Park, Hae Ryoun; Lee, Ji Hye

    2016-02-01

    Copy number variations at multiple chromosomal loci, including 16p11.2, have recently been implicated in the pathogenesis of autism spectrum disorder (ASD), a neurodevelopmental disease that affects 1~3% of children worldwide. The aim of this study was to investigate the roles of human genes at the 16p11.2 loci in synaptic development using Drosophila larval neuromuscular junctions (NMJ), a well-established model synapse with stereotypic innervation patterns. We conducted a preliminary genetic screen based on RNA interference in combination with the GAL4-UAS system, followed by mutational analyses. Our result indicated that disruption of klp68D, a gene closely related to human KIF22, caused ectopic innervations of axon branches forming type III boutons in muscle 13, along with less frequent re-routing of other axon branches. In addition, mutations in klp64D, of which gene product forms Kinesin-2 complex with KLP68D, led to similar targeting errors of type III axons. Mutant phenotypes were at least partially reproduced by knockdown of each gene via RNA interference. Taken together, our data suggest the roles of Kinesin-2 proteins, including KLP68D and KLP64D, in ensuring proper synaptic wiring. PMID:26924931

  1. Serpent, Suppressor of Hairless and U-shaped are crucial regulators of hedgehog niche expression and prohemocyte maintenance during Drosophila larval hematopoiesis

    PubMed Central

    Tokusumi, Yumiko; Tokusumi, Tsuyoshi; Stoller-Conrad, Jessica; Schulz, Robert A.

    2010-01-01

    The lymph gland is a specialized organ for hematopoiesis, utilized during larval development in Drosophila. This tissue is composed of distinct cellular domains populated by blood cell progenitors (the medullary zone), niche cells that regulate the choice between progenitor quiescence and hemocyte differentiation [the posterior signaling center (PSC)], and mature blood cells of distinct lineages (the cortical zone). Cells of the PSC express the Hedgehog (Hh) signaling molecule, which instructs cells within the neighboring medullary zone to maintain a hematopoietic precursor state while preventing hemocyte differentiation. As a means to understand the regulatory mechanisms controlling Hh production, we characterized a PSC-active transcriptional enhancer that drives hh expression in supportive niche cells. Our findings indicate that a combination of positive and negative transcriptional inputs program the precise PSC expression of the instructive Hh signal. The GATA factor Serpent (Srp) is essential for hh activation in niche cells, whereas the Suppressor of Hairless [Su(H)] and U-shaped (Ush) transcriptional regulators prevent hh expression in blood cell progenitors and differentiated hemocytes. Furthermore, Srp function is required for the proper differentiation of niche cells. Phenotypic analyses also indicated that the normal activity of all three transcriptional regulators is essential for maintaining the progenitor population and preventing premature hemocyte differentiation. Together, these studies provide mechanistic insights into hh transcriptional regulation in hematopoietic progenitor niche cells, and demonstrate the requirement of the Srp, Su(H) and Ush proteins in the control of niche cell differentiation and blood cell precursor maintenance. PMID:20876645

  2. Drosophila Homolog of Human KIF22 at the Autism-Linked 16p11.2 Loci Influences Synaptic Connectivity at Larval Neuromuscular Junctions

    PubMed Central

    Park, Sang Mee; Littleton, J. Troy; Park, Hae Ryoun

    2016-01-01

    Copy number variations at multiple chromosomal loci, including 16p11.2, have recently been implicated in the pathogenesis of autism spectrum disorder (ASD), a neurodevelopmental disease that affects 1~3% of children worldwide. The aim of this study was to investigate the roles of human genes at the 16p11.2 loci in synaptic development using Drosophila larval neuromuscular junctions (NMJ), a well-established model synapse with stereotypic innervation patterns. We conducted a preliminary genetic screen based on RNA interference in combination with the GAL4-UAS system, followed by mutational analyses. Our result indicated that disruption of klp68D, a gene closely related to human KIF22, caused ectopic innervations of axon branches forming type III boutons in muscle 13, along with less frequent re-routing of other axon branches. In addition, mutations in klp64D, of which gene product forms Kinesin-2 complex with KLP68D, led to similar targeting errors of type III axons. Mutant phenotypes were at least partially reproduced by knockdown of each gene via RNA interference. Taken together, our data suggest the roles of Kinesin-2 proteins, including KLP68D and KLP64D, in ensuring proper synaptic wiring. PMID:26924931

  3. Identification of Inhibitory Premotor Interneurons Activated at a Late Phase in a Motor Cycle during Drosophila Larval Locomotion

    PubMed Central

    Itakura, Yuki; Kohsaka, Hiroshi; Ohyama, Tomoko; Zlatic, Marta

    2015-01-01

    Rhythmic motor patterns underlying many types of locomotion are thought to be produced by central pattern generators (CPGs). Our knowledge of how CPG networks generate motor patterns in complex nervous systems remains incomplete, despite decades of work in a variety of model organisms. Substrate borne locomotion in Drosophila larvae is driven by waves of muscular contraction that propagate through multiple body segments. We use the motor circuitry underlying crawling in larval Drosophila as a model to try to understand how segmentally coordinated rhythmic motor patterns are generated. Whereas muscles, motoneurons and sensory neurons have been well investigated in this system, far less is known about the identities and function of interneurons. Our recent study identified a class of glutamatergic premotor interneurons, PMSIs (period-positive median segmental interneurons), that regulate the speed of locomotion. Here, we report on the identification of a distinct class of glutamatergic premotor interneurons called Glutamatergic Ventro-Lateral Interneurons (GVLIs). We used calcium imaging to search for interneurons that show rhythmic activity and identified GVLIs as interneurons showing wave-like activity during peristalsis. Paired GVLIs were present in each abdominal segment A1-A7 and locally extended an axon towards a dorsal neuropile region, where they formed GRASP-positive putative synaptic contacts with motoneurons. The interneurons expressed vesicular glutamate transporter (vGluT) and thus likely secrete glutamate, a neurotransmitter known to inhibit motoneurons. These anatomical results suggest that GVLIs are premotor interneurons that locally inhibit motoneurons in the same segment. Consistent with this, optogenetic activation of GVLIs with the red-shifted channelrhodopsin, CsChrimson ceased ongoing peristalsis in crawling larvae. Simultaneous calcium imaging of the activity of GVLIs and motoneurons showed that GVLIs’ wave-like activity lagged behind that of

  4. Immediate and punitive impact of mechanosensory disturbance on olfactory behaviour of larval Drosophila

    PubMed Central

    Saumweber, Timo; Cano, Carmen; Klessen, Juliane; Eichler, Katharina; Fendt, Markus; Gerber, Bertram

    2014-01-01

    ABSTRACT The ability to respond to and to learn about mechanosensory disturbance is widespread among animals. Using Drosophila larvae, we describe how the frequency of mechanosensory disturbance (‘buzz’) affects three aspects of behaviour: free locomotion, innate olfactory preference, and potency as a punishment. We report that (i) during 2–3 seconds after buzz onset the larvae slowed down and then turned, arguably to escape this situation; this was seen for buzz frequencies of 10, 100, and 1000 Hz, (ii) innate olfactory preference was reduced when tested in the presence of the buzz; this effect was strongest for the 100 Hz frequency, (iii) after odour-buzz associative training, we observed escape from the buzz-associated odour; this effect was apparent for 10 and 100, but not for 1000 Hz. We discuss the multiple behavioural effects of mechanosensation and stress that the immediate effects on locomotion and the impact as punishment differ in their frequency-dependence. Similar dissociations between immediate, reflexive behavioural effects and reinforcement potency were previously reported for sweet, salty and bitter tastants. It should be interesting to see how these features map onto the organization of sensory, ascending pathways. PMID:25260919

  5. Immediate and punitive impact of mechanosensory disturbance on olfactory behaviour of larval Drosophila.

    PubMed

    Saumweber, Timo; Cano, Carmen; Klessen, Juliane; Eichler, Katharina; Fendt, Markus; Gerber, Bertram

    2014-01-01

    The ability to respond to and to learn about mechanosensory disturbance is widespread among animals. Using Drosophila larvae, we describe how the frequency of mechanosensory disturbance ('buzz') affects three aspects of behaviour: free locomotion, innate olfactory preference, and potency as a punishment. We report that (i) during 2-3 seconds after buzz onset the larvae slowed down and then turned, arguably to escape this situation; this was seen for buzz frequencies of 10, 100, and 1000 Hz, (ii) innate olfactory preference was reduced when tested in the presence of the buzz; this effect was strongest for the 100 Hz frequency, (iii) after odour-buzz associative training, we observed escape from the buzz-associated odour; this effect was apparent for 10 and 100, but not for 1000 Hz. We discuss the multiple behavioural effects of mechanosensation and stress that the immediate effects on locomotion and the impact as punishment differ in their frequency-dependence. Similar dissociations between immediate, reflexive behavioural effects and reinforcement potency were previously reported for sweet, salty and bitter tastants. It should be interesting to see how these features map onto the organization of sensory, ascending pathways. PMID:25260919

  6. The impact of odor–reward memory on chemotaxis in larval Drosophila

    PubMed Central

    Schleyer, Michael; Reid, Samuel F.; Pamir, Evren; Saumweber, Timo; Paisios, Emmanouil; Davies, Alexander

    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) where to direct a turn. We first report how odor-source intensities modulate these decisions to bring about higher levels of chemotactic performance for higher odor-source intensities during innate chemotaxis. We then examine whether the same modulations are responsible for alterations of chemotactic performance by learned odor “valence” (understood throughout as level of attractiveness). We find that run speed (i) is neither modulated by the innate nor by the learned valence of an odor. Turn rate (ii), however, is modulated by both: the higher the innate or learned valence of the odor, the less often larvae turn whenever heading toward the odor source, and the more often they turn when heading away. Likewise, turning direction (iii) is modulated concordantly by innate and learned valence: turning is biased more strongly toward the odor source when either innate or learned valence is high. Using numerical simulations, we show that a modulation of both turn rate and of turning direction is sufficient to account for the empirically found differences in preference scores across experimental conditions. Our results suggest that innate and learned valence organize adaptive olfactory search behavior by their summed effects on turn rate and turning direction, but not on run speed. This work should aid studies into the neural mechanisms by which memory impacts specific aspects of behavior. PMID:25887280

  7. Resource Limitation, Controphic Ostracod Density and Larval Mosquito Development

    PubMed Central

    Rowbottom, Raylea; Carver, Scott; Barmuta, Leon A.; Weinstein, Philip; Foo, Dahlia; Allen, Geoff R.

    2015-01-01

    Aquatic environments can be restricted with the amount of available food resources especially with changes to both abiotic and biotic conditions. Mosquito larvae, in particular, are sensitive to changes in food resources. Resource limitation through inter-, and intra-specific competition among mosquitoes are known to affect both their development and survival. However, much less is understood about the effects of non-culicid controphic competitors (species that share the same trophic level). To address this knowledge gap, we investigated and compared mosquito larval development, survival and adult size in two experiments, one with different densities of non-culicid controphic conditions and the other with altered resource conditions. We used Aedes camptorhynchus, a salt marsh breeding mosquito and a prominent vector for Ross River virus in Australia. Aedes camptorhynchus usually has few competitors due to its halo-tolerance and distribution in salt marshes. However, sympatric ostracod micro-crustaceans often co-occur within these salt marshes and can be found in dense populations, with field evidence suggesting exploitative competition for resources. Our experiments demonstrate resource limiting conditions caused significant increases in mosquito developmental times, decreased adult survival and decreased adult size. Overall, non-culicid exploitation experiments showed little effect on larval development and survival, but similar effects on adult size. We suggest that the alterations of adult traits owing to non-culicid controphic competition has potential to extend to vector-borne disease transmission. PMID:26558896

  8. Resource Limitation, Controphic Ostracod Density and Larval Mosquito Development.

    PubMed

    Rowbottom, Raylea; Carver, Scott; Barmuta, Leon A; Weinstein, Philip; Foo, Dahlia; Allen, Geoff R

    2015-01-01

    Aquatic environments can be restricted with the amount of available food resources especially with changes to both abiotic and biotic conditions. Mosquito larvae, in particular, are sensitive to changes in food resources. Resource limitation through inter-, and intra-specific competition among mosquitoes are known to affect both their development and survival. However, much less is understood about the effects of non-culicid controphic competitors (species that share the same trophic level). To address this knowledge gap, we investigated and compared mosquito larval development, survival and adult size in two experiments, one with different densities of non-culicid controphic conditions and the other with altered resource conditions. We used Aedes camptorhynchus, a salt marsh breeding mosquito and a prominent vector for Ross River virus in Australia. Aedes camptorhynchus usually has few competitors due to its halo-tolerance and distribution in salt marshes. However, sympatric ostracod micro-crustaceans often co-occur within these salt marshes and can be found in dense populations, with field evidence suggesting exploitative competition for resources. Our experiments demonstrate resource limiting conditions caused significant increases in mosquito developmental times, decreased adult survival and decreased adult size. Overall, non-culicid exploitation experiments showed little effect on larval development and survival, but similar effects on adult size. We suggest that the alterations of adult traits owing to non-culicid controphic competition has potential to extend to vector-borne disease transmission. PMID:26558896

  9. Hypomorphic Mutations in the Larval Photokinesis a (Lpha) Gene Have Stage-Specific Effects on Visual System Function in Drosophila Melanogaster

    PubMed Central

    Gordesky-Gold, B.; Warrick, J. M.; Bixler, A.; Beasley, J. E.; Tompkins, L.

    1995-01-01

    Of the many genes that are expressed in the visual system of Drosophila melanogaster adults, some affect larval vision. However, with the exception of one X-linked mutation, no genes that have larval-specific effects on visual system structure or function have previously been reported. We describe the isolation and characterization of two mutant alleles that define the larval photokinesis A (lphA) gene, one allele of which is associated with a P-element insertion at cytogenetic locus 8E1-10. Larvae that express lphA mutations are, like normal animals, negatively photokinetic, but they are less responsive to white light than lphA+ controls. Larvae that are heterozygous in trans for a mutant lphA allele and a deficiency that uncovers the lphA locus are blind, which indicates that the mutant allele is hypomorphic. lphA larvae respond normally to odorants and taste stimuli. Moreover, the lphA mutations do not affect adult flies' fast phototaxis or visually driven aspects of male sexual behavior, and electroretinograms recorded from the compound eyes of lphA/deficiency heterozygotes and lphA(1)/lphA(2) females are normal. These observations suggest that the lphA gene affects a larval-specific aspect of visual system function. PMID:7789764

  10. Torso, a Drosophila receptor tyrosine kinase, plays a novel role in the larval fat body in regulating insulin signaling and body growth.

    PubMed

    Jun, Jong Woo; Han, Gangsik; Yun, Hyun Myoung; Lee, Gang Jun; Hyun, Seogang

    2016-08-01

    Torso is a receptor tyrosine kinase whose localized activation at the termini of the Drosophila embryo is mediated by its ligand, Trunk. Recent studies have unveiled a second function of Torso in the larval prothoracic gland (PG) as the receptor for the prothoracicotropic hormone, which triggers pupariation. As such, inhibition of Torso in the PG prolongs the larval growth period, thereby increasing the final pupa size. Here, we report that Torso also acts in the larval fat body, regulating body size in a manner opposite from that of Torso in PG. We confirmed the expression of torso mRNA in the larval fat body and its reduction by RNA interference (RNAi). Fat body-specific knockdown of torso, by either of the two independent RNAi transgenes, significantly decreased the final pupal size. We found that torso knockdown suppresses insulin/target of rapamycin (TOR) signaling in the fat body, as confirmed by repression of Akt and S6K. Notably, the decrease in insulin/TOR signaling and decrease of pupal size induced by the knockdown of torso were rescued by the expression of a constitutively active form of the insulin receptor or by the knockdown of FOXO. Our study revealed a novel role for Torso in the fat body with respect to regulation of insulin/TOR signaling and body size. This finding exemplifies the contrasting effects of the same gene expressed in two different organs on organismal physiology. PMID:27126913

  11. Hypomorphic mutations in the larval photokinesis A (lphA) gene have stage-specific effects on visual system function in Drosophila melanogaster

    SciTech Connect

    Gordesky-Gold, B.; Warrick, J.M.; Bixler, A.

    1995-04-01

    Of the many genes that are expressed in the visual system of Drosophila melanogaster adults, some affect larval vision. However, with the exception of one X-linked mutation, no genes that have larval-specific effects on visual system structure or function have previously been reported. We describe the isolation and characterization of two mutant alleles that define the larval photokinesis A (lphA) gene, one allele of which is associated with a P-element insertion at cytogenetic locus 8E1-10. Larvae that express lphA mutations are, like normal animals, negatively photokinetic, but they are less responsive to white light than lphA+ controls. Larvae that are heterozygous in trans for a mutant allele and a deficiency that uncovers the lphA locus are blind, which indicates that the mutant allele is hypomorphic. lphA larvae respond normally to odorants and taste stimuli. Moreover, the lphA mutations do not affect adult flies` fast phototaxis or visually driven aspects of male sexual behavior, and electroretinograms recorded from the compound eyes of lphA/deficiency heterozygotes and lphA{sup 1}/lphA{sup 2} females are normal. These observations suggest that the lphA gene affects a larval-specific aspect of visual system function. 27 refs., 2 figs., 4 tabs.

  12. Proteomic analysis through larval development of Solea senegalensis flatfish.

    PubMed

    Chicano-Gálvez, Eduardo; Asensio, Esther; Cañavate, José Pedro; Alhama, José; López-Barea, Juan

    2015-12-01

    The post-embryonic development of the Senegalese sole, Solea senegalensis, a flatfish of growing interest in fisheries and aquaculture, is associated with drastic morpho-physiological changes during metamorphosis. Although in the last two decades knowledge on sole culture has notably increased, especially in Southern Europe, its progress was restricted due to lack of methods to control reproduction, improve larval quality and increase juvenile disease resistance. A limited knowledge of the physiological, molecular and genetic mechanisms involved is at the base of such limitation. A proteomic study was carried out to explore the molecular events that occur during S. senegalensis ontogenesis. Protein expression changes were monitored in larvae from 5 to 21 dph by combining 2DE and protein identification with de novo MS/MS sequencing. An average of 6177 ± 282 spots was resolved in 2DE gels. Hierarchical cluster analysis of the 705 selected spots grouped them in eight patterns. Thirty-four proteins were identified and assigned biological functions including structure, metabolism highlighting energy metabolism, transport, protein folding, stress response, chromatin organization and regulation of gene expression. These changes provide a sequential description of the molecular events associated with the biochemical and biological transformations that occur during sole larval development. PMID:26365915

  13. Biased gene expression in early honeybee larval development

    PubMed Central

    2013-01-01

    Background Female larvae of the honeybee (Apis mellifera) develop into either queens or workers depending on nutrition. This nutritional stimulus triggers different developmental trajectories, resulting in adults that differ from each other in physiology, behaviour and life span. Results To understand how these trajectories are established we have generated a comprehensive atlas of gene expression throughout larval development. We found substantial differences in gene expression between worker and queen-destined larvae at 6 hours after hatching. Some of these early changes in gene expression are maintained throughout larval development, indicating that caste-specific developmental trajectories are established much earlier than previously thought. Within our gene expression data we identified processes that potentially underlie caste differentiation. Queen-destined larvae have higher expression of genes involved in transcription, translation and protein folding early in development with a later switch to genes involved in energy generation. Using RNA interference, we were able to demonstrate that one of these genes, hexamerin 70b, has a role in caste differentiation. Both queen and worker developmental trajectories are associated with the expression of genes that have alternative splice variants, although only a single variant of a gene tends to be differentially expressed in a given caste. Conclusions Our data, based on the biases in gene expression early in development together with published data, supports the idea that caste development in the honeybee consists of two phases; an initial biased phase of development, where larvae can still switch to the other caste by differential feeding, followed by commitment to a particular developmental trajectory. PMID:24350621

  14. Transcription factor broad suppresses precocious development of adult structures during larval-pupal metamorphosis in the red flour beetle, Tribolium castaneum.

    PubMed

    Parthasarathy, R; Tan, A; Bai, H; Palli, Subba R

    2008-01-01

    Broad (br), a transcription factor containing the Broad-Tramtrack-Bric-a-brac (BTB) and zinc finger domains was shown to mediate 20-hydroxyecdysone (20E) action and pupal development in Drosophila melanogaster and Manduca sexta. We determined the key roles of br during larval-pupal metamorphosis using RNA interference (RNAi) in a coleopteran insect, Tribolium castaneum. Two major peaks of T. castaneum broad (Tcbr) mRNA, one peak at the end of feeding stage prior to the larvae entering the quiescent stage and another peak during the quiescent stage were detected in the whole body and midgut tissue dissected from staged insects. Expression of br during the final instar larval stage is essential for successful larval-pupal metamorphosis, because, RNAi-mediated knock-down of Tcbr during this stage derailed larval-pupal metamorphosis and produced insects that showed larval, pupal and adult structures. Tcbr dsRNA injected into the final instar larvae caused reduction in the mRNA levels of genes known to be involved in 20E action (EcRA, E74 and E75B). Tcbr dsRNA injected into the final instar larvae also caused an increase in the mRNA levels of JH-response genes (JHE and Kr-h1b). Knock-down of Tcbr expression also affected 20E-mediated remodeling of midgut during larval-pupal metamorphosis. These data suggest that the expression of Tcbr during the final instar larval stage promotes pupal program while suppressing the larval and adult programs ensuring a transitory pupal stage in holometabolous insects. PMID:18083350

  15. Functions of the Nonsense-Mediated mRNA Decay Pathway in Drosophila Development

    PubMed Central

    Metzstein, Mark M; Krasnow, Mark A

    2006-01-01

    Nonsense-mediated mRNA decay (NMD) is a cellular surveillance mechanism that degrades transcripts containing premature translation termination codons, and it also influences expression of certain wild-type transcripts. Although the biochemical mechanisms of NMD have been studied intensively, its developmental functions and importance are less clear. Here, we describe the isolation and characterization of Drosophila “photoshop” mutations, which increase expression of green fluorescent protein and other transgenes. Mapping and molecular analyses show that photoshop mutations are loss-of-function mutations in the Drosophila homologs of NMD genes Upf1, Upf2, and Smg1. We find that Upf1 and Upf2 are broadly active during development, and they are required for NMD as well as for proper expression of dozens of wild-type genes during development and for larval viability. Genetic mosaic analysis shows that Upf1 and Upf2 are required for growth and/or survival of imaginal cell clones, but this defect can be overcome if surrounding wild-type cells are eliminated. By contrast, we find that the PI3K-related kinase Smg1 potentiates but is not required for NMD or for viability, implying that the Upf1 phosphorylation cycle that is required for mammalian and Caenorhabditis elegans NMD has a more limited role during Drosophila development. Finally, we show that the SV40 3′ UTR, present in many Drosophila transgenes, targets the transgenes for regulation by the NMD pathway. The results establish that the Drosophila NMD pathway is broadly active and essential for development, and one critical function of the pathway is to endow proliferating imaginal cells with a competitive growth advantage that prevents them from being overtaken by other proliferating cells. PMID:17196039

  16. The black-pearl gene of Drosophila defines a novel conserved protein family and is required for larval growth and survival.

    PubMed

    Becker, S; Gehrsitz, A; Bork, P; Buchner, S; Buchner, E

    2001-01-10

    Using a transposon insertion line of the Drosophila Genome Project we have cloned the black-pearl gene (blp), analyzed cDNA clones, generated various mutants, and characterized their phenotypes. The blp gene codes for a protein of 15.7 kDa calculated molecular weight that has been conserved from yeast to plants and mammals with high homology. A domain of these new proteins shows distant similarity to DnaJ domains indicating a functionally relevant interaction with other proteins. The P element insertion in line P1539 lies within the 5' untranslated leader of the black-pearl gene. Flies homozygous for this insertion are semi-lethal, escapers produce very few offspring and show melanotic inclusions in the hemocoel ('black pearls') similar to various melanotic 'tumor' mutants. Two small deletions confined to the blp gene and two EMS-induced mutations are homozygous lethal. These null mutants appear normal up to a prolonged first instar larval stage but fail to grow and die. Thus in Drosophila the blp gene is specifically required for larval growth. The evolutionary conservation in both unicellular and multicellular organisms suggests for the new protein family described here a fundamental role in cell growth. PMID:11179663

  17. Drosophila brain development: closing the gap between a macroarchitectural and microarchitectural approach.

    PubMed

    Cardona, A; Saalfeld, S; Tomancak, P; Hartenstein, V

    2009-01-01

    Neurobiologists address neural structure, development, and function at the level of "macrocircuits" (how different brain compartments are interconnected; what overall pattern of activity they produce) and at the level of "microcircuits" (how connectivity and physiology of individual neurons and their processes within a compartment determine the functional output of this compartment). Work in our lab aims at reconstructing the developing Drosophila brain at both levels. Macrocircuits can be approached conveniently by reconstructing the pattern of brain lineages, which form groups of neurons whose projections form cohesive fascicles interconnecting the compartments of the larval and adult brain. The reconstruction of microcircuits requires serial section electron microscopy, due to the small size of terminal neuronal processes and their synaptic contacts. Because of the amount of labor that traditionally comes with this approach, very little is known about microcircuitry in brains across the animal kingdom. Many of the problems of serial electron microscopy reconstruction are now solvable with digital image recording and specialized software for both image acquisition and postprocessing. In this chapter, we introduce our efforts to reconstruct the small Drosophila larval brain and discuss our results in light of the published data on neuropile ultrastructure in other animal taxa. PMID:20028843

  18. Drosophila brain development: Closing the gap between a macroarchitectural and microarchitectural approach

    PubMed Central

    Cardona, Albert; Saalfeld, Stefan; Tomancak, Pavel; Hartenstein, Volker

    2014-01-01

    Neurobiologists address neural structure, development and function at the level of “macrocircuits” (how are different brain compartments interconnected, what overall pattern of activity do they produce), and at the level of “microcircuits” (how does connectivity and physiology of individual neurons and their processes within a compartment determine the functional output of this compartment). Work in our lab aims at reconstructing the developing Drosophila brain at both levels. Macrocircuits can be approached conveniently by reconstructing the pattern of brain lineages, which form groups of neurons whose projections form cohesive fascicles interconnecting the compartments of the larval and adult brain. The reconstruction of microcircuits requires serial section electron microscopy, due to the small size of terminal neuronal processes and their synaptic contacts. Because of the amount of labor that traditionally comes with this approach, very little is known about microcircuitry in brains across the animal kingdom. Many of the problems of serial EM reconstruction is now solvable with digital image recording and specialized software for both image acquisition and post-processing. In this paper we introduce our efforts to reconstruct the small Drosophila larval brain, and discuss our results in light of the published data on neuropile ultrastructure in other animal taxa. PMID:20028843

  19. 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

  20. 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.

  1. Embryonic and larval development of Brycon amazonicus (SPIX & AGASSIZ, 1829).

    PubMed

    Nakauth, A C S Sampaio; Villacorta-Correa, M A; Figueiredo, M R; Bernardino, G; França, J M

    2016-02-01

    The objective of this study was to describe the embryonic and larval development of Brycon amazonicus, featuring the main events up to 50 hours after fertilization (AF). The material was provided by the Aquaculture Training, Technology and Production Center, Presidente Figueiredo (AM). The characterization was based on stereomicroscopic examination of the morphology of eggs, embryos and larvae and comparison with the literature. Matrinxã eggs are free, transparent, and spherical, with a perivitelline space of 0.56 ± 0.3 mm. The successive divisions give rise to cells with 64 blastomeres during the first hour AF. The gastrula stage, beginning 02 h 40 min AF, was characterized by progressive regression cells and the formation of the embryonic axis, leading to differentiation of the head and tail 05 h 30 min AF. From 06 to 09 h AF the somites, notochord, otic and optic vesicles and otoliths were observed, in addition to heart rate and the release of the tail. The larvae hatched at 10 h 30 min AF (29.9 °C), with a total length of 3.56 ± 0.46 mm. Between 19 and 30 h AF, we observed 1) pigmentation and gut formation, 2) branchial arches, 3) pectoral fins, 4) a mouth opening and 5) teeth. Cannibalism was initiated earlier (34 h AF) which was associated with rapid yolk absorption (more than 90% until 50 h AF), signaling the need for an exogenous nutritional source. The environmental conditions (especially temperature) influenced the time course of some events throughout the embryonic and larval development, suggesting the need for further studies on this subject. PMID:26909629

  2. Loss of a Clueless-dGRASP complex results in ER stress and blocks Integrin exit from the perinuclear endoplasmic reticulum in Drosophila larval muscle

    PubMed Central

    Wang, Zong-Heng; Rabouille, Catherine; Geisbrecht, Erika R.

    2015-01-01

    Drosophila Clueless (Clu) and its conserved orthologs are known for their role in the prevention of mitochondrial clustering. Here, we uncover a new role for Clu in the delivery of integrin subunits in muscle tissue. In clu mutants, αPS2 integrin, but not βPS integrin, abnormally accumulates in a perinuclear endoplasmic reticulum (ER) subdomain, a site that mirrors the endogenous localization of Clu. Loss of components essential for mitochondrial distribution do not phenocopy the clu mutant αPS2 phenotype. Conversely, RNAi knockdown of the Drosophila Golgi reassembly and stacking protein GRASP55/65 (dGRASP) recapitulates clu defects, including the abnormal accumulation of αPS2 and larval locomotor activity. Both Clu and dGRASP proteins physically interact and loss of Clu displaces dGRASP from ER exit sites, suggesting that Clu cooperates with dGRASP for the exit of αPS2 from a perinuclear subdomain in the ER. We also found that Clu and dGRASP loss of function leads to ER stress and that the stability of the ER exit site protein Sec16 is severely compromised in the clu mutants, thus explaining the ER accumulation of αPS2. Remarkably, exposure of clu RNAi larvae to chemical chaperones restores both αPS2 delivery and functional ER exit sites. We propose that Clu together with dGRASP prevents ER stress and therefore maintains Sec16 stability essential for the functional organization of perinuclear early secretory pathway. This, in turn, is essential for integrin subunit αPS2 ER exit in Drosophila larval myofibers. PMID:25862246

  3. Ascorbic Acid Influences the Development and Immunocompetence of Larval Heliothis virescens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report that ascorbic acid, which is known to be a free radical scavenger, to be important not only in insect development but also in larval resistance to baculovirus infection. We sequentially elevated the ascorbic acid content in diet and evaluated the effect on larval H. virescens development ...

  4. Morphological identification and development of neurite in Drosophila ventral nerve cord neuropil.

    PubMed

    Gan, Guangming; Lv, Huihui; Xie, Wei

    2014-01-01

    In Drosophila, ventral nerve cord (VNC) occupies most of the larval central nervous system (CNS). However, there is little literature elaborating upon the specific types and growth of neurites as defined by their structural appearance in Drosophila larval VNC neuropil. Here we report the ultrastructural development of different types VNC neurites in ten selected time points in embryonic and larval stages utilizing transmission electron microscopy. There are four types of axonal neurites as classified by the type of vesicular content: clear vesicle (CV) neurites have clear vesicles and some T-bar structures; Dense-core vesicle (DV) neurites have dense-core vesicles and without T-bar structures; Mixed vesicle (MV) neurites have mixed vesicles and some T-bar structures; Large vesicle (LV) neurites are dominated by large, translucent spherical vesicles but rarely display T-bar structures. We found dramatic remodeling in CV neurites which can be divided into five developmental phases. The neurite is vacuolated in primary (P) phase, they have mitochondria, microtubules or big dark vesicles in the second (S) phase, and they contain immature synaptic features in the third (T) phase. The subsequent bifurcate (B) phase appears to undergo major remodeling with the appearance of the bifurcation or dendritic growth. In the final mature (M) phase, high density of commensurate synaptic vesicles are distributed around T-bar structures. There are four kinds of morphological elaboration of the CVI neurite sub-types. First, new neurite produces at the end of axon. Second, new neurite bubbles along the axon. Third, the preexisting neurite buds and develops into several neurites. The last, the bundled axons form irregularly shape neurites. Most CVI neurites in M phase have about 1.5-3 µm diameter, they could be suitable to analyze their morphology and subcellular localization of specific proteins by light microscopy, and they could serve as a potential model in CNS in vivo development

  5. Effects of Spaceflight on Drosophila Neural Development

    NASA Technical Reports Server (NTRS)

    Keshishian, Haig S.

    1997-01-01

    The major goal from the animal side, however, has been achieved, namely to develop Drosophila lines where we can assay individual neuromuscular endings directly without dissection. This was achieved by means of using the GAL4-UAS system, where we have succeeded in establishing stocks of flies where the key neuromuscular connections can be assayed directly in undissected larvae by means of the expression of endogenously fluorescent reporters in the specific motor endings. The green fluorescent protein (GFP) as a reporter allows scoring of neural anatomy en-masse in whole mount using fluorescent microscopy without the need for either dissection or specific labeling. Two stocks have been developed. The first, which we developed first, uses the S65T mutant form, which has a dramatically brighter expression than the native protein. This animal will use GAL4 drivers with expression under the control of the elav gene, and which will ensure expression in all neurons of the embryo and larva. The second transgenic animal we have developed is of a novel kind, and makes use of dicistronic design, so that two copies of the protein will be expressed per insert. We have also developed a tricistronic form, but this has not yet been transformed into flies, and we do not imagine that this third line will be ready in time for the flight.

  6. Correlated Evolution between Mode of Larval Development and Habitat in Muricid Gastropods

    PubMed Central

    Pappalardo, Paula; Rodríguez-Serrano, Enrique; Fernández, Miriam

    2014-01-01

    Abstract Larval modes of development affect evolutionary processes and influence the distribution of marine invertebrates in the ocean. The decrease in pelagic development toward higher latitudes is one of the patterns of distribution most frequently discussed in marine organisms (Thorson's rule), which has been related to increased larval mortality associated with long pelagic durations in colder waters. However, the type of substrate occupied by adults has been suggested to influence the generality of the latitudinal patterns in larval development. To help understand how the environment affects the evolution of larval types we evaluated the association between larval development and habitat using gastropods of the Muricidae family as a model group. To achieve this goal, we collected information on latitudinal distribution, sea water temperature, larval development and type of substrate occupied by adults. We constructed a molecular phylogeny for 45 species of muricids to estimate the ancestral character states and to assess the relationship between traits using comparative methods in a Bayesian framework. Our results showed high probability for a common ancestor of the muricids with nonpelagic (and nonfeeding) development, that lived in hard bottoms and cold temperatures. From this ancestor, a pelagic feeding larva evolved three times, and some species shifted to warmer temperatures or sand bottoms. The evolution of larval development was not independent of habitat; the most probable evolutionary route reconstructed in the analysis of correlated evolution showed that type of larval development may change in soft bottoms but in hard bottoms this change is highly unlikely. Lower sea water temperatures were associated with nonpelagic modes of development, supporting Thorson's rule. We show how environmental pressures can favor a particular mode of larval development or transitions between larval modes and discuss the reacquisition of feeding larva in muricids

  7. Correlated evolution between mode of larval development and habitat in muricid gastropods.

    PubMed

    Pappalardo, Paula; Rodríguez-Serrano, Enrique; Fernández, Miriam

    2014-01-01

    Larval modes of development affect evolutionary processes and influence the distribution of marine invertebrates in the ocean. The decrease in pelagic development toward higher latitudes is one of the patterns of distribution most frequently discussed in marine organisms (Thorson's rule), which has been related to increased larval mortality associated with long pelagic durations in colder waters. However, the type of substrate occupied by adults has been suggested to influence the generality of the latitudinal patterns in larval development. To help understand how the environment affects the evolution of larval types we evaluated the association between larval development and habitat using gastropods of the Muricidae family as a model group. To achieve this goal, we collected information on latitudinal distribution, sea water temperature, larval development and type of substrate occupied by adults. We constructed a molecular phylogeny for 45 species of muricids to estimate the ancestral character states and to assess the relationship between traits using comparative methods in a Bayesian framework. Our results showed high probability for a common ancestor of the muricids with nonpelagic (and nonfeeding) development, that lived in hard bottoms and cold temperatures. From this ancestor, a pelagic feeding larva evolved three times, and some species shifted to warmer temperatures or sand bottoms. The evolution of larval development was not independent of habitat; the most probable evolutionary route reconstructed in the analysis of correlated evolution showed that type of larval development may change in soft bottoms but in hard bottoms this change is highly unlikely. Lower sea water temperatures were associated with nonpelagic modes of development, supporting Thorson's rule. We show how environmental pressures can favor a particular mode of larval development or transitions between larval modes and discuss the reacquisition of feeding larva in muricids gastropods

  8. Collective synchronization of divisions in Drosophila development

    NASA Astrophysics Data System (ADS)

    Vergassola, Massimo

    Mitoses in the early development of most metazoans are rapid and synchronized across the entire embryo. While diffusion is too slow, in vitro experiments have shown that waves of the cell-cycle regulator Cdk1 can transfer information rapidly across hundreds of microns. However, the signaling dynamics and the physical properties of chemical waves during embryonic development remain unclear. We develop FRET biosensors for the activity of Cdk1 and the checkpoint kinase Chk1 in Drosophila embryos and exploit them to measure waves in vivo. We demonstrate that Cdk1 chemical waves control mitotic waves and that their speed is regulated by the activity of Cdk1 during the S-phase (and not mitosis). We quantify the progressive slowdown of the waves with developmental cycles and identify its underlying control mechanism by the DNA replication checkpoint through the Chk1/Wee1 pathway. The global dynamics of the mitotic signaling network illustrates a novel control principle: the S-phase activity of Cdk1 regulates the speed of the mitotic wave, while the Cdk1 positive feedback ensures an invariantly rapid onset of mitosis. Mathematical modeling captures the speed of the waves and predicts a fundamental distinction between the S-phase Cdk1 trigger waves and the mitotic phase waves, which is illustrated by embryonic ablation experiments. In collaboration with Victoria Deneke1, Anna Melbinger2, and Stefano Di Talia1 1 Department of Cell Biology, Duke University Medical Center 2 Department of Physics, University of California San Diego.

  9. Effects of Disinfectants on Larval Development of Ascaris suum Eggs.

    PubMed

    Oh, Ki-Seok; Kim, Geon-Tae; Ahn, Kyu-Sung; Shin, Sung-Shik

    2016-02-01

    The objective of this study was to evaluate the effects of several different commercial disinfectants on the embryogenic development of Ascaris suum eggs. A 1-ml aliquot of each disinfectant was mixed with approximately 40,000 decorticated or intact A. suum eggs in sterile tubes. After each treatment time (at 0.5, 1, 5, 10, 30, and 60 min), disinfectants were washed away, and egg suspensions were incubated at 25˚C in distilled water for development of larvae inside. At 3 weeks of incubation after exposure, ethanol, methanol, and chlorohexidin treatments did not affect the larval development of A. suum eggs, regardless of their concentration and treatment time. Among disinfectants tested in this study, 3% cresol, 0.2% sodium hypochlorite and 0.02% sodium hypochlorite delayed but not inactivated the embryonation of decorticated eggs at 3 weeks of incubation, because at 6 weeks of incubation, undeveloped eggs completed embryonation regardless of exposure time, except for 10% povidone iodine. When the albumin layer of A. suum eggs remained intact, however, even the 10% povidone iodine solution took at least 5 min to reasonably inactivate most eggs, but never completely kill them with even 60 min of exposure. This study demonstrated that the treatment of A. suum eggs with many commercially available disinfectants does not affect the embryonation. Although some disinfectants may delay or stop the embryonation of A. suum eggs, they can hardly kill them completely. PMID:26951988

  10. Effects of Disinfectants on Larval Development of Ascaris suum Eggs

    PubMed Central

    Oh, Ki-Seok; Kim, Geon-Tae; Ahn, Kyu-Sung; Shin, Sung-Shik

    2016-01-01

    The objective of this study was to evaluate the effects of several different commercial disinfectants on the embryogenic development of Ascaris suum eggs. A 1-ml aliquot of each disinfectant was mixed with approximately 40,000 decorticated or intact A. suum eggs in sterile tubes. After each treatment time (at 0.5, 1, 5, 10, 30, and 60 min), disinfectants were washed away, and egg suspensions were incubated at 25˚C in distilled water for development of larvae inside. At 3 weeks of incubation after exposure, ethanol, methanol, and chlorohexidin treatments did not affect the larval development of A. suum eggs, regardless of their concentration and treatment time. Among disinfectants tested in this study, 3% cresol, 0.2% sodium hypochlorite and 0.02% sodium hypochlorite delayed but not inactivated the embryonation of decorticated eggs at 3 weeks of incubation, because at 6 weeks of incubation, undeveloped eggs completed embryonation regardless of exposure time, except for 10% povidone iodine. When the albumin layer of A. suum eggs remained intact, however, even the 10% povidone iodine solution took at least 5 min to reasonably inactivate most eggs, but never completely kill them with even 60 min of exposure. This study demonstrated that the treatment of A. suum eggs with many commercially available disinfectants does not affect the embryonation. Although some disinfectants may delay or stop the embryonation of A. suum eggs, they can hardly kill them completely. PMID:26951988

  11. Central brain postembryonic development in Drosophila: implication of genes expressed at the interhemispheric junction.

    PubMed

    Boquet, I; Hitier, R; Dumas, M; Chaminade, M; Préat, T

    2000-01-01

    Postembryonic brain development of Drosophila has become recently a subject of intense investigations. In particular, the linotte (lio) mutants display strong structural defects in the mushroom bodies and the central complex. The Lio kinase is expressed in a glial structure at the interhemispheric junction of late larval and young pupal brain. With the aim of identifying new genes involved in the formation of adult central brain structures, 821 enhancer-trap Gal4 lines were generated and screened for late larval expression. We identified 167 lines showing expression at or near the interhemispheric junction of third-instar larval brain, an area from which the central complex differentiates. Adult brains from 104 of these 167 lines were analyzed through paraffin sections. This secondary screen allowed the recovery of five central brain mutants. Of 89 control lines showing various patterns of expression excluding the interhemispheric junction, only one anatomical mutant was isolated. These six mutations, which have been thoroughly characterized, affect the midline area of the adult brain with phenotypes of split central complex structures and/or fused mushroom body lobes. This work opens the way for further analysis of the molecular and cellular events involved in central brain reorganization during metamorphosis. PMID:10623899

  12. Patterns of growth, axonal extension and axonal arborization of neuronal lineages in the developing Drosophila brain.

    PubMed

    Larsen, Camilla; Shy, Diana; Spindler, Shana R; Fung, Siaumin; Pereanu, Wayne; Younossi-Hartenstein, Amelia; Hartenstein, Volker

    2009-11-15

    The Drosophila central brain is composed of approximately 100 paired lineages, with most lineages comprising 100-150 neurons. Most lineages have a number of important characteristics in common. Typically, neurons of a lineage stay together as a coherent cluster and project their axons into a coherent bundle visible from late embryo to adult. Neurons born during the embryonic period form the primary axon tracts (PATs) that follow stereotyped pathways in the neuropile. Apoptotic cell death removes an average of 30-40% of primary neurons around the time of hatching. Secondary neurons generated during the larval period form secondary axon tracts (SATs) that typically fasciculate with their corresponding primary axon tract. SATs develop into the long fascicles that interconnect the different compartments of the adult brain. Structurally, we distinguish between three types of lineages: PD lineages, characterized by distinct, spatially separate proximal and distal arborizations; C lineages with arborizations distributed continuously along the entire length of their tract; D lineages that lack proximal arborizations. Arborizations of many lineages, in particular those of the PD type, are restricted to distinct neuropile compartments. We propose that compartments are "scaffolded" by individual lineages, or small groups thereof. Thereby, the relatively small number of primary neurons of each primary lineage set up the compartment map in the late embryo. Compartments grow during the larval period simply by an increase in arbor volume of primary neurons. Arbors of secondary neurons form within or adjacent to the larval compartments, resulting in smaller compartment subdivisions and additional, adult specific compartments. PMID:19538956

  13. Chitosan/siRNA nanoparticle targeting demonstrates a requirement for single-minded during larval and pupal olfactory system development of the vector mosquito Aedes aegypti

    PubMed Central

    2014-01-01

    Background Essentially nothing is known about the genetic regulation of olfactory system development in vector mosquitoes, which use olfactory cues to detect blood meal hosts. Studies in Drosophila melanogaster have identified a regulatory matrix of transcription factors that controls pupal/adult odorant receptor (OR) gene expression in olfactory receptor neurons (ORNs). However, it is unclear if transcription factors that function in the D. melanogaster regulatory matrix are required for OR expression in mosquitoes. Furthermore, the regulation of OR expression during development of the larval olfactory system, which is far less complex than that of pupae/adults, is not well understood in any insect, including D. melanogaster. Here, we examine the regulation of OR expression in the developing larval olfactory system of Aedes aegypti, the dengue vector mosquito. Results A. aegypti bears orthologs of eight transcription factors that regulate OR expression in D. melanogaster pupae/adults. These transcription factors are expressed in A. aegypti larval antennal sensory neurons, and consensus binding sites for these transcription factors reside in the 5’ flanking regions of A. aegypti OR genes. Consensus binding sites for Single-minded (Sim) are located adjacent to over half the A. aegypti OR genes, suggesting that this transcription factor functions as a major regulator of mosquito OR expression. To functionally test this hypothesis, chitosan/siRNA nanoparticles were used to target sim during larval olfactory development. These experiments demonstrated that Sim positively regulates expression of a large subset of OR genes, including orco, the obligate co-receptor in the assembly and function of heteromeric OR/Orco complexes. Decreased innervation of the antennal lobe was also noted in sim knockdown larvae. These OR expression and antennal lobe defects correlated with a larval odorant tracking behavioral defect. OR expression and antennal lobe defects were also

  14. The proprotein convertase amontillado (amon) is required during Drosophila pupal development.

    PubMed

    Rayburn, Lowell Y M; Rhea, Jeanne; Jocoy, Steven R; Bender, Michael

    2009-09-01

    Peptide hormones governing many developmental processes are generated via endoproteolysis of inactive precursor molecules by a family of subtilisin-like proprotein convertases (SPCs). We previously identified mutations in the Drosophila amontillado (amon) gene, a homolog of the vertebrate neuroendocrine-specific Prohormone Convertase 2 (PC2) gene, and showed that amon is required during embryogenesis, early larval development, and larval molting. Here, we define amon requirements during later developmental stages using a conditional rescue system and find that amon is required during pupal development for head eversion, leg and wing disc extension, and abdominal differentiation. Immuno-localization experiments show that amon protein is expressed in a subset of central nervous system cells but does not co-localize with peptide hormones known to elicit molting behavior, suggesting the involvement of novel regulatory peptides in this process. The amon protein is expressed in neuronal cells that innervate the corpus allatum and corpora cardiaca of the ring gland, an endocrine organ which is the release site for many key hormonal signals. Expression of amon in a subset of these cell types using the GAL4/UAS system in an amon mutant background partially rescues larval molting and growth. Our results show that amon is required for pupal development and identify a subset of neuronal cell types in which amon function is sufficient to rescue developmental progression and growth defects shown by amon mutants. The results are consistent with a model that the amon protein acts to proteolytically process a diverse suite of peptide hormones that coordinate larval and pupal growth and development. PMID:19559693

  15. The proprotein convertase amontillado (amon) is required during Drosophila pupal development

    PubMed Central

    Rayburn, Lowell Y.M.; Rhea, Jeanne; Jocoy, Steven R.; Bender, Michael

    2009-01-01

    Peptide hormones governing many developmental processes are generated via endoproteolysis of inactive precursor molecules by a family of subtilisin-like proprotein convertases (SPCs). We previously identified mutations in the Drosophila amontillado (amon) gene, a homolog of the vertebrate neuroendocrine-specific Prohormone Convertase 2 (PC2) gene, and showed that amon is required during embryogenesis, early larval development, and larval molting. Here, we define amon requirements during later developmental stages using a conditional rescue system and find that amon is required during pupal development for head eversion, leg and wing disc extension, and abdominal differentiation. Immuno-localization experiments show that amon protein is expressed in a subset of central nervous system cells but does not co-localize with peptide hormones known to elicit molting behavior, suggesting the involvement of novel regulatory peptides in this process. The amon protein is expressed in neuronal cells that innervate the corpus allatum and corpora cardiaca of the ring gland, an endocrine organ which is the release site for many key hormonal signals. Expression of amon in a subset of these cell types using the GAL4/UAS system in an amon mutant background partially rescues larval molting and growth. Our results show that amon is required for pupal development and identify a subset of neuronal cell types in which amon function is sufficient to rescue developmental progression and growth defects shown by amon mutants. The results are consistent with a model that the amon protein acts to proteolytically process a diverse suite of peptide hormones that coordinate larval and pupal growth and development. PMID:19559693

  16. Elevated major ion concentrations inhibit larval mayfly growth and development.

    PubMed

    Johnson, Brent R; Weaver, Paul C; Nietch, Christopher T; Lazorchak, James M; Struewing, Katherine A; Funk, David H

    2015-01-01

    Anthropogenic disturbances, including those from developing energy resources, can alter stream chemistry significantly by elevating total dissolved solids. Field studies have indicated that mayflies (Order Ephemeroptera) are particularly sensitive to high total dissolved solids. In the present study, the authors measured 20-d growth and survivorship of larval Neocloeon triangulifer exposed to a gradient of brine salt (mixed NaCl and CaCl2 ) concentrations. Daily growth rates were reduced significantly in all salt concentrations above the control (363 µS cm(-1) ) and larvae in treatments with specific conductance >812 µS cm(-1) were in comparatively earlier developmental stages (instars) at the end of the experiment. Survivorship declined significantly when specific conductance was >1513 µS cm(-1) and the calculated 20-d 50% lethal concentration was 2866 µS cm(-1) . The present study's results provide strong experimental evidence that elevated ion concentrations similar to those observed in developing energy resources, such as oil and gas drilling or coal mining, can adversely affect sensitive aquatic insect species. PMID:25307284

  17. Effects of Synthetic Diets Enriched in Specific Nutrients on Drosophila Development, Body Fat, and Lifespan

    PubMed Central

    Reis, Tânia

    2016-01-01

    Gene-diet interactions play a crucial but poorly understood role in susceptibility to obesity. Accordingly, the development of genetically tractable model systems to study the influence of diets in obesity-prone genetic backgrounds is a focus of current research. Here I present a modified synthetic Drosophila diet optimized for timely larval development, a stage dedicated to energy storage. Specifically increasing the levels of individual macronutrients–carbohydrate, lipid, or protein–resulted in markedly different organismal effects. A high-carbohydrate diet adversely affected the timing of development, size, early lifespan and body fat. Strikingly, quadrupling the amount of dietary lipids had none of these effects. Diets rich in protein appeared to be the most beneficial, as larvae developed faster, with no change in size, into long-lived adults. I believe this synthetic diet will significantly facilitate the study of gene-diet interactions in organismal energy balance. PMID:26741692

  18. Effects of Synthetic Diets Enriched in Specific Nutrients on Drosophila Development, Body Fat, and Lifespan.

    PubMed

    Reis, Tânia

    2016-01-01

    Gene-diet interactions play a crucial but poorly understood role in susceptibility to obesity. Accordingly, the development of genetically tractable model systems to study the influence of diets in obesity-prone genetic backgrounds is a focus of current research. Here I present a modified synthetic Drosophila diet optimized for timely larval development, a stage dedicated to energy storage. Specifically increasing the levels of individual macronutrients-carbohydrate, lipid, or protein-resulted in markedly different organismal effects. A high-carbohydrate diet adversely affected the timing of development, size, early lifespan and body fat. Strikingly, quadrupling the amount of dietary lipids had none of these effects. Diets rich in protein appeared to be the most beneficial, as larvae developed faster, with no change in size, into long-lived adults. I believe this synthetic diet will significantly facilitate the study of gene-diet interactions in organismal energy balance. PMID:26741692

  19. Larval development of Culex quinquefasciatus in water with low to moderate.

    PubMed

    Noori, Navideh; Lockaby, B Graeme; Kalin, Latif

    2015-12-01

    Population growth and urbanization have increased the potential habitats, and consequently the abundance of Culex quinquefasciatus, the southern house mosquito, a vector of West Nile Virus in urban areas. Water quality is critical in larval habitat distribution and in providing microbial food resources for larvae. A mesocosm experiment was designed to demonstrate which specific components of water chemistry are conducive to larval Culex mosquitoes. Dose-response relationships between larval development and NO3 , NH4 , and PO4 concentrations in stream water were developed through this experiment to describe the isolated effects of each nutrient on pre-adult development. The emergence pattern of Culex mosquitoes was found to be strongly related to certain nutrients, and results showed that breeding sites with higher PO4 or NO3 concentrations had higher larval survival rates. High NO3 concentrations favor the development of male mosquitoes and suppress the development of female mosquitoes, but those adult females that do emerge develop faster in containers with high NO3 levels compared to the reference group. The addition of PO4 in the absence of nitrogen sources to the larval habitat slowed larval development, however, it took fewer days for larvae to reach the pupal stage in containers with combinations of NO3 and PO4 or NH4 and PO4 nutrients. Results from this study may bolster efforts to control WNV in urban landscapes by exploring water quality conditions of Culex larval habitats that produce adult mosquitoes. PMID:26611953

  20. Effect of ace inhibitors and TMOF on growth, development, and trypsin activity of larval Spodoptera littoralis.

    PubMed

    Lemeire, Els; Borovsky, Dov; Van Camp, John; Smagghe, Guy

    2008-12-01

    Angiotensin converting enzyme (ACE) is a zinc metallopeptidase capable of cleaving dipeptide or dipeptideamide moieties at the C-terminal end of peptides. ACE is present in the hemolymph and reproductive tissues of insects. The presence of ACE in the hemolymph and its broad substrate specificity suggests an important role in processing of bioactive peptides. This study reports the effects of ACE inhibitors on larval growth in the cotton leafworm Spodoptera littoralis. Feeding ACE inhibitors ad lib decreased the growth rate, inhibited ACE activity in the larval hemolymph, and down-regulated trypsin activity in the larval gut. These results indicate that S. littoralis ACE may influence trypsin biosynthesis in the larval gut by interacting with a trypsin-modulating oostatic factor (TMOF). Injecting third instar larvae with a combination of Aea-TMOF and the ACE inhibitor captopril, down-regulated trypsin biosynthesis in the larval gut indicating that an Aea-TMOF gut receptor analogue could be present. Injecting captopril and enalapril into newly molted fifth instar larvae stopped larval feeding and decreased weight gain. Together, these results indicate that ACE inhibitors are efficacious in stunting larval growth and ACE plays an important role in larval growth and development. PMID:18949805

  1. Effect of three larval diets on larval development and male sexual performance of Anopheles gambiae s.s.

    PubMed

    Yahouédo, Gildas A; Djogbénou, Luc; Saïzonou, Jacques; Assogba, Bénoît S; Makoutodé, Michel; Gilles, Jeremie R L; Maïga, Hamidou; Mouline, Karine; Soukou, Bhonna K; Simard, Frédéric

    2014-04-01

    Population replacement/elimination strategies based on mass-release of sterile or otherwise genetically modified (male) mosquitoes are being considered in order to expand the malaria vector control arsenal on the way to eradication. A challenge in this context, is to produce male mosquitoes that will be able to compete and mate with wild females more efficiently than their wild counterparts, i.e. high fitness males. This study explored the effect of three larval food diets developed by the International Atomic Energy Agency on the overall fitness and mating performance of male Anopheles gambiae s.s. mosquitoes (Kisumu strain). Larval development (pupation and emergence rate, development time) was monitored, and adult wing length and energy reserves at emergence (i.e. lipids, sugars, glycogen and proteins) were measured. Male sexual performance was assessed through an insemination test whereby one male and 10 virgin females were maintained together in the same cage in order to record the number of inseminated females per 24h. Our results show that males reared on Diets 2 and 3 performed best during larval development. Males provided with treatment 2.2 had a shorter development time and performed best in insemination tests. However, these males had the lowest overall lifespan, suggesting a trade-off between longevity and sexual performances which needs to be taken into consideration when planning release. The results from this work were discussed in the context of sterile insect techniques or genetic control methods which is today one of the strategy in the overall mosquito control and elimination efforts. PMID:24291460

  2. Drosophila larval to pupal switch under nutrient stress requires IP3R/Ca2+ signalling in glutamatergic interneurons

    PubMed Central

    Jayakumar, Siddharth; Richhariya, Shlesha; Reddy, O Venkateswara; Texada, Michael J; Hasan, Gaiti

    2016-01-01

    Neuronal circuits are known to integrate nutritional information, but the identity of the circuit components is not completely understood. Amino acids are a class of nutrients that are vital for the growth and function of an organism. Here, we report a neuronal circuit that allows Drosophila larvae to overcome amino acid deprivation and pupariate. We find that nutrient stress is sensed by the class IV multidendritic cholinergic neurons. Through live calcium imaging experiments, we show that these cholinergic stimuli are conveyed to glutamatergic neurons in the ventral ganglion through mAChR. We further show that IP3R-dependent calcium transients in the glutamatergic neurons convey this signal to downstream medial neurosecretory cells (mNSCs). The circuit ultimately converges at the ring gland and regulates expression of ecdysteroid biosynthetic genes. Activity in this circuit is thus likely to be an adaptation that provides a layer of regulation to help surpass nutritional stress during development. DOI: http://dx.doi.org/10.7554/eLife.17495.001 PMID:27494275

  3. INFLUENCE OF AN INSECT GROWTH REGULATOR ON THE LARVAL DEVELOPMENT OF AN ESTUARINE SHRIMP

    EPA Science Inventory

    The influence of methoprene, an insect growth regulator used in mosquito control, on larval development of the estuarine grass shrimp (Palaemonetes pugio) was examined in the laboratory. o grass shrimp larvae successfully completed metamorphosis when continuously exposed to 1000 ...

  4. A Flippase-Mediated GAL80/GAL4 Intersectional Resource for Dissecting Appendage Development in Drosophila.

    PubMed

    Smith, Brittany N; Ghazanfari, Arash M; Bohm, Rudolf A; Welch, William P; Zhang, Bing; Masly, John P

    2015-10-01

    Drosophila imaginal discs provide an ideal model to study processes important for cell signaling and cell specification, tissue differentiation, and cell competition during development. One challenge to understanding genetic control of cellular processes and cell interactions is the difficulty in effectively targeting a defined subset of cells in developing tissues in gene manipulation experiments. A recently developed Flippase-induced intersectional GAL80/GAL4 repression method incorporates several gene manipulation technologies in Drosophila to enable such fine-scale dissection in neural tissues. In particular, this approach brings together existing GAL4 transgenes, newly developed enhancer-trap flippase transgenes, and GAL80 transgenes flanked by Flippase recognition target sites. The combination of these tools enables gene activation/repression in particular subsets of cells within a GAL4 expression pattern. Here, we expand the utility of a large collection of these enhancer-trap flippase transgenic insertion lines by characterizing their expression patterns in third larval instar imaginal discs. We screened 521 different enhancer-trap flippase lines and identified 28 that are expressed in imaginal tissues, including two transgenes that show sex-specific expression patterns. Using a line that expresses Flippase in the wing imaginal disc, we demonstrate the utility of this intersectional approach for studying development by knocking down gene expression of a key member of the planar cell polarity pathway. The results of our experiments show that these enhancer-trap flippase lines enable fine-scale manipulation in imaginal discs. PMID:26276385

  5. Worniu, a Snail family zinc-finger protein, is required for brain development in Drosophila.

    PubMed

    Ashraf, Shovon I; Ganguly, Atish; Roote, John; Ip, Y Tony

    2004-10-01

    The Snail family of zinc-finger transcriptional repressors is essential for morphogenetic cell movements, mesoderm formation, and neurogenesis during embryonic development. These proteins also control cell cycle, cell death, and cancer progression. In Drosophila, three members of this protein family, Snail, Escargot, and Worniu, have essential but redundant functions in asymmetric cell division of neuroblasts. In addition, Snail is critical for early mesoderm formation and Escargot is required for maintaining diploidy in wing imaginal disc cells. In this report, we demonstrate that Worniu plays a role in brain development. We show that alleles of the l(2)35Da complementation group are mutants of worniu. The developing larvae of these mutant alleles fail to shorten their brainstems. The brain phenotype, as well as the lethality, of these mutants can be rescued by worniu transgenes. Moreover, RNAi experiments targeting the worniu transcript show the same nonshortening phenotype in larval brains. worniu is expressed in the neuroblasts of brain hemispheres and ventral ganglions. The results suggest that the loss of Worniu function within the neuroblasts ultimately causes the larval brainstem to fail to go through shortening during development. PMID:15366015

  6. Larval development of the subantarctic king crabs Lithodes santolla and Paralomis granulosa reared in the laboratory

    NASA Astrophysics Data System (ADS)

    Calcagno, J. A.; Anger, K.; Lovrich, G. A.; Thatje, S.; Kaffenberger, A.

    2004-02-01

    The larval development and survival in the two subantarctic lithodid crabs Lithodes santolla (Jaquinot) and Paralomis granulosa (Molina) from the Argentine Beagle Channel were studied in laboratory cultures. In L. santolla, larval development lasted about 70 days, passing through three zoeal stages and the megalopa stage, with a duration of approximately 4, 7, 11 and 48 days, respectively. The larval development in P. granulosa is more abbreviated, comprising only two zoeal stages and the megalopa stage, with 6, 11 and 43 days' duration, respectively. In both species, we tested for effects of presence versus absence of food (Artemia nauplii) on larval development duration and survival rate. In P. granulosa, we also studied effects of different rearing conditions, such as individual versus mass cultures, as well as aerated versus unaerated cultures. No differences in larval development duration and survival were observed between animals subjected to those different rearing conditions. The lack of response to the presence or absence of potential food confirms, in both species, a complete lecithotrophic mode of larval development. Since lithodid crabs are of high economic importance in the artisanal fishery in the southernmost parts of South America, the knowledge of optimal rearing conditions for lithodid larvae is essential for future attempts at repopulating the collapsing natural stocks off Tierra del Fuego.

  7. Changes in boron concentration during development and ageing of Drosophila and effect of dietary boron on life span.

    PubMed

    Massie, H R; Whitney, S J; Aiello, V R; Sternick, S M

    1990-03-31

    Total boron concentrations in Drosophila changed during development and ageing. The highest concentration of boron was found during the egg stage followed by a decline during the larval stages. Newly emerged flies contained 35.5 ppm boron. During the adult stage the boron concentration increased by 52% by 9 weeks of age. Adding excess dietary boron during the adult stage decreased the median life span by 69% at 0.01 M sodium borate and by 21% at 0.001 M sodium borate. Lower concentrations gave small but significant increases in life span. Supplementing a very low boron diet with 0.00025 M sodium borate improved life span by 9.5%. The boron contents of young and old mouse tissues were similar to those of Drosophila and human samples. We conclude that moderate levels of dietary boron may have a general protective effect in biological systems. The mechanism of this effect at present remains unknown. PMID:2325439

  8. Larval and metamorphic development of the foregut and proboscis in the caenogastropod Marsenina (Lamellaria) stearnsii.

    PubMed

    Page, L R

    2002-05-01

    The specialized, postmetamorphic feeding structures of predatory caenogastropods evolved by changes to an ancestral caenogastropod developmental program that generated a planktotrophic larval stage followed by a herbivorous postmetamorphic stage. As part of a program of comparative studies aimed at reconstructing these developmental changes, I studied the development of the postmetamorphic feeding system of Marsenina stearnsii using histological sections for light microscopy and scanning and transmission electron microscopy. The feeding system of this species has two very different designs during ontogeny. The larval system uses ciliary effectors to capture and ingest microalgae, whereas the juvenile/adult system includes a proboscis, jaws, and radular apparatus for predation on ascidian zooids. The postmetamorphic foregut begins to develop during the early larval phase, but the anlagen does not interfere with larval feeding because it develops as an increasingly elaborate outpocketing from the ventral wall of the larval esophagus. At metamorphosis, an opening is created in the anterior tip of the prospective, postmetamorphic buccal cavity and the margins of this opening anneal with the metamorphically remodeled lips of the larval mouth. This process exposes the jaws, which differentiate within the buccal cavity prior to metamorphosis. As a working hypothesis, I suggest that rupture of the buccal cavity to the outside at metamorphosis was selected as a mechanism to allow precocious development of jaws in species where jaws enhanced feeding performance by young juveniles. The larval esophagus of M. stearnsii appears to be completely destroyed at metamorphosis. Larval esophageal cells have distinctive apical characteristics (cilia, blebbed microvilli, stacks of lamellae within the glycocalyx) and no cells having this signature persist through metamorphosis. Development of the proboscis and proboscis sac, which begins prior to metamorphosis, conforms to previous

  9. Regulation of apoptosis of rbf mutant cells during Drosophila development

    PubMed Central

    Tanaka-Matakatsu, Miho; Xu, Jinhua; Cheng, Leping; Du, Wei

    2008-01-01

    Inactivation of the retinoblastoma gene Rb leads to defects in cell proliferation, differentiation, or apoptosis, depending on specific cell or tissue types. To gain insights into the genes that can modulate the consequences of Rb inactivation, we carried out a genetic screen in Drosophila to identify mutations that affected apoptosis induced by inactivation of the Retinoblastoma-family protein (rbf) and identified a mutation that blocked apoptosis induced by rbf. We found this mutation to be a new allele of head involution defective (hid) and showed that hid expression is deregulated in rbf mutant cells in larval imaginal discs. We identified an enhancer that regulates hid expression in response to developmental cues as well as to radiation and demonstrated that this hid enhancer is directly repressed by RBF through an E2F binding site. These observations indicate that apoptosis of rbf mutant cells is mediated by an upregulation of hid. Finally, we showed that bantam, a miRNA that regulates hid translation, is expressed in the interommatidial cells in the larval eye discs and modulates the survival of rbf mutant cells. PMID:19100727

  10. Adenylate kinase isozyme 2 is essential for growth and development of Drosophila melanogaster.

    PubMed

    Fujisawa, Koichi; Murakami, Ryutaro; Horiguchi, Taigo; Noma, Takafumi

    2009-05-01

    Adenylate kinases are phylogenetically widespread, highly conserved, and involved in energy metabolism and energy transfer. Of these, adenylate kinase (AK) isozyme 2 is uniquely localized in the mitochondrial intermembrane space and its physiological role remains largely unknown. In this study, we selected Drosophila melanogaster to analyze its role in vivo. AK isozyme cDNAs were cloned and their gene expressions were characterized in D. melanogaster. The deduced amino acid sequences contain highly conserved motifs for P-loop, NMP binding, and LID domains of AKs. In addition, the effects of AK2 gene knockout on phenotype of AK2 mutants were examined using P-element technology. Although homozygous AK2 mutated embryos developed without any visible defects, their growth ceased and they died before reaching the third instar larval stage. Maternally provided AK2 mRNA was detected in fertilized eggs, and weak AK2 activity was observed in first and second instar larvae of the homozygous AK2 mutants, suggesting that maternally provided AK2 is sufficient for embryonic development. Disappearance of AK2 activity during larval stages resulted in growth arrest and eventual death. These results demonstrate that AK2 plays a critical role in adenine nucleotide metabolism in the mitochondrial intermembrane space and is essential for growth in D. melanogaster. PMID:19416704

  11. Exploration of the "larval pool": development and ground-truthing of a larval transport model off leeward Hawai'i.

    PubMed

    Wren, Johanna L K; Kobayashi, Donald R

    2016-01-01

    Most adult reef fish show site fidelity thus dispersal is limited to the mobile larval stage of the fish, and effective management of such species requires an understanding of the patterns of larval dispersal. In this study, we assess larval reef fish distributions in the waters west of the Big Island of Hawai'i using both in situ and model data. Catches from Cobb midwater trawls off west Hawai'i show that reef fish larvae are most numerous in offshore waters deeper than 3,000 m and consist largely of pre-settlement Pomacanthids, Acanthurids and Chaetodontids. Utilizing a Lagrangian larval dispersal model, we were able to replicate the observed shore fish distributions from the trawl data and we identified the 100 m depth strata as the most likely depth of occupancy. Additionally, our model showed that for larval shore fish with a pelagic larval duration longer than 40 days there was no significant change in settlement success in our model. By creating a general additive model (GAM) incorporating lunar phase and angle we were able to explain 67.5% of the variance between modeled and in situ Acanthurid abundances. We took steps towards creating a predictive larval distribution model that will greatly aid in understanding the spatiotemporal nature of the larval pool in west Hawai'i, and the dispersal of larvae throughout the Hawaiian archipelago. PMID:26855873

  12. Development of the Drosophila entero-endocrine lineage and its specification by the Notch signaling pathway.

    PubMed

    Takashima, Shigeo; Adams, Katrina L; Ortiz, Paola A; Ying, Chong T; Moridzadeh, Rameen; Younossi-Hartenstein, Amelia; Hartenstein, Volker

    2011-05-15

    In this paper we have investigated the developmental-genetic steps that shape the entero-endocrine system of Drosophila melanogaster from the embryo to the adult. The process starts in the endoderm of the early embryo where precursors of endocrine cells and enterocytes of the larval midgut, as well as progenitors of the adult midgut, are specified by a Notch signaling-dependent mechanism. In a second step that occurs during the late larval period, enterocytes and endocrine cells of a transient pupal midgut are selected from within the clusters of adult midgut progenitors. As in the embryo, activation of the Notch pathway triggers enterocyte differentiation and inhibits cells from further proliferation or choosing the endocrine fate. The third step of entero-endocrine cell development takes place at a mid-pupal stage. Before this time point, the epithelial layer destined to become the adult midgut is devoid of endocrine cells. However, precursors of the intestinal midgut stem cells (pISCs) are already present. After an initial phase of symmetric divisions which causes an increase in their own population size, pISCs start to spin off cells that become postmitotic and express the endocrine fate marker, Prospero. Activation of Notch in pISCs forces these cells into an enterocyte fate. Loss of Notch function causes an increase in the proliferatory activity of pISCs, as well as a higher ratio of Prospero-positive cells. PMID:21382366

  13. Development of the Drosophila entero-endocrine lineage and its specification by the Notch signaling pathway

    PubMed Central

    Takashima, Shigeo; Adams, Katrina L.; Ortiz, Paola A.; Ying, Chong T.; Moridzadeh, Rameen; Younossi-Hartenstein, Amelia; Hartenstein, Volker

    2013-01-01

    In this paper we have investigated the developmental-genetic steps that shape the entero-endocrine system of Drosophila melanogaster from the embryo to the adult. The process starts in the endoderm of the early embryo where precursors of endocrine cells and enterocytes of the larval midgut, as well as progenitors of the adult midgut, are specified by a Notch signaling-dependent mechanism. In a second step that occurs during the late larval period, enterocytes and endocrine cells of a transient pupal midgut are selected from within the clusters of adult midgut progenitors. As in the embryo, activation of the Notch pathway triggers enterocyte differentiation, and inhibits cells from further proliferation or choosing the endocrine fate. The third step of entero-endocrine cell development takes place at a mid-pupal stage. Before this time point, the epithelial layer destined to become the adult midgut is devoid of endocrine cells. However, precursors of the intestinal midgut stem cells (pISCs) are already present. After an initial phase of symmetric divisions which causes an increase in their own population size, pISCs start to spin off cells that become postmitotic and express the endocrine fate marker, Prospero. Activation of Notch in pISCs forces these cells into an enterocyte fate. Loss of Notch function causes an increase in the proliferatory activity of pISCs, as well as a higher ratio of Prospero-positive cells. PMID:21382366

  14. Tracheal development in the Drosophila brain is constrained by glial cells

    PubMed Central

    Pereanu, Wayne; Spindler, Shana; Cruz, Luis; Hartenstein, Volker

    2007-01-01

    The Drosophila brain is tracheated by the cerebral trachea, a branch of the first segmental trachea of the embryo. During larval stages the cerebral trachea splits into several main (primary) branches that grow around the neuropile, forming a perineuropilar tracheal plexus (PNP) at the neuropile surface. Five primary tracheal branches whose spatial relationship to brain compartments is relatively invariant can be distinguished, although the exact trajectories and branching pattern of the brain tracheae is surprisingly variable. Immuno-histochemical and electron microscopic demonstrate that all brain tracheae grow in direct contact with the glial cell processes that surround the neuropile. To investigate the effect of glia on tracheal development, embryos and larvae lacking glial cells as a result of a genetic mutation or a directed ablation were analyzed. In these animals, the tracheal branching pattern was highly abnormal. In particular, the number of secondary branches entering the central neuropile was increased. Wild type larvae possess only two central tracheae, typically associated with the mushroom body and the antenno-cerebral tract. In larvae lacking glial cells, six to ten tracheal branches penetrate the neuropile in a variable pattern. This finding indicates that glia-derived signals constrained tracheal growth in the Drosophila brain and restrict the number of branches entering the neuropile. PMID:17046740

  15. Patterns of growth, axonal extension and axonal arborization of neuronal lineages in the developing Drosophila brain

    PubMed Central

    Larsen, Camilla; Shy, Diana; Spindler, Shana R.; Fung, Siaumin; Pereanu, Wayne; Younossi -Hartenstein, Amelia; Hartenstein, Volker

    2009-01-01

    The Drosophila central brain is composed of approximately 100 paired lineages, with most lineages comprising 100–150 neurons. Most lineages have a number of important characteristics in common. Typically, neurons of a lineage stay together as a coherent cluster and project their axons into a coherent bundle visible from late embryo to adult. Neurons born during the embryonic period form the primary axon tracts (PATs) that follow stereotyped pathways in the neuropile. Apoptotic cell death removes an average of 30–40% of primary neurons around the time of hatching. Secondary neurons generated during the larval period form secondary axon tracts (SATs) that typically fasciculate with their corresponding primary axon tract. SATs develop into the long fascicles that interconnect the different compartments of the adult brain. Structurally, we distinguish between three types of lineages: PD lineages, characterized by distinct, spatially separate proximal and distal arborizations; C lineages with arborizations distributed continuously along the entire length of their tract; D lineages that lack proximal arborizations. Arborizations of many lineages, in particular those of the PD type, are restricted to distinct neuropile compartments. We propose that compartments are ‘scaffolded” by individual lineages, or small groups thereof. Thereby, the relatively small number of primary neurons of each primary lineage set up the compartment map in the late embryo. Compartments grow during the larval period simply by an increase in arbor volume of primary neurons. Arbors of secondary neurons form within or adjacent to the larval compartments, resulting in smaller compartment subdivisions and additional, adult specific compartments. PMID:19538956

  16. Genetic architecture of olfactory behavior in Drosophila melanogaster: differences and similarities across development

    PubMed Central

    Lavagnino, N.J.; Arya, G.H.; Korovaichuk, A.; Fanara, J.J.

    2013-01-01

    In the holometabolous insect Drosophila melanogaster, genetic, physiological and anatomical aspects of olfaction are well known in the adult stage, while larval stages olfactory behavior has received some attention it has been less studied than its adult counterpart. Most of these studies focus on olfactory receptors (Or) genes that produce peripheral odor recognition. In this paper, through a loss-of-function screen using P-element inserted lines and also by means of expression analyses of larval olfaction candidate genes, we extended the uncovering of the genetic underpinnings of D. melanogaster larval olfactory behavior by demonstrating that larval olfactory behavior is, in addition to Or genes, orchestrated by numerous genes with diverse functions. Also, our results points out that the genetic architecture of olfactory behavior in D. melanogaster presents a dynamic and changing organization across environments and ontogeny. PMID:23563598

  17. Effects of ammonia on fertilization, development, and larval survival in the Northern Pacific asteroid, Asterias amurensis.

    PubMed

    Lee, Chang-Hoon; Sung, Chan-Gyoung; Moon, Seong-Dae; Lee, Jong-Hyeon

    2013-07-01

    For developing a complementary test organism to sea urchin during winter in Korea, sensitivities of sperm, embryo, and larvae of Asterias amurensis to un-ionized ammonia were evaluated. The EC₅₀s (Mean ± SD, n = 3) for fertilization and development were 169 ± 62 and 70 ± 19 μg/L, respectively. The 48, 72, and 96-h LC₅₀s for larval survival were 1,674 ± 583, 498 ± 221, and 336 ± 107 μg/L, respectively. The sensitivities of fertilization, development, and larval survival tests with A. amurensis are higher than or comparable to those of sea urchin and other taxonomic groups. Therefore, fertilization, development, and larval survival tests using A. amurensis are suitable for assessing pore water toxicity of marine sediments in Korea. PMID:23674221

  18. The Drosophila Hand gene is required for remodeling of the developing adult heart and midgut during metamorphosis

    PubMed Central

    Lo, Patrick C.H.; Zaffran, Stéphane; Sénatore, Sébastien; Frasch, Manfred

    2007-01-01

    The Hand proteins of the bHLH family of transcriptional factors play critical roles in vertebrate cardiogenesis. In Drosophila, the single orthologous Hand gene is expressed in the developing embryonic dorsal vessel (heart), lymph glands, circular visceral musculature, and a subset of CNS cells. We demonstrate that the absence of Hand activity causes semilethality during the early larval instars. The dorsal vessel and midgut musculature are unaffected in null mutant embryos, but in a large fraction the lymph glands are missing. However, homozygous adult flies lacking Hand possess morphologically abnormal dorsal vessels characterized by a disorganized myofibrillar structure, reduced systolic and diastolic diameter, abnormal heartbeat contractions, and suffer from premature lethality. In addition, their midguts are highly deformed; in the most severe cases, there is midgut blockage and a massive excess of ectopic peritrophic membrane tubules exiting a rupture in an anterior midgut bulge. Nevertheless, the visceral musculature appears to be relatively normal. Based on these phenotypes, we conclude that the expression of the Drosophila Hand gene in the dorsal vessel and circular visceral muscles is mainly required during pupal stages, when Hand participates in the proper hormone-dependent remodeling of the larval aorta into the adult heart and in the normal morphogenesis of the adult midgut endoderm during metamorphosis. PMID:17904115

  19. Modeling larval connectivity of the Atlantic surfclams within the Middle Atlantic Bight: Model development, larval dispersal and metapopulation connectivity

    NASA Astrophysics Data System (ADS)

    Zhang, Xinzhong; Haidvogel, Dale; Munroe, Daphne; Powell, Eric N.; Klinck, John; Mann, Roger; Castruccio, Frederic S.

    2015-02-01

    To study the primary larval transport pathways and inter-population connectivity patterns of the Atlantic surfclam, Spisula solidissima, a coupled modeling system combining a physical circulation model of the Middle Atlantic Bight (MAB), Georges Bank (GBK) and the Gulf of Maine (GoM), and an individual-based surfclam larval model was implemented, validated and applied. Model validation shows that the model can reproduce the observed physical circulation patterns and surface and bottom water temperature, and recreates the observed distributions of surfclam larvae during upwelling and downwelling events. The model results show a typical along-shore connectivity pattern from the northeast to the southwest among the surfclam populations distributed from Georges Bank west and south along the MAB shelf. Continuous surfclam larval input into regions off Delmarva (DMV) and New Jersey (NJ) suggests that insufficient larval supply is unlikely to be the factor causing the failure of the population to recover after the observed decline of the surfclam populations in DMV and NJ from 1997 to 2005. The GBK surfclam population is relatively more isolated than populations to the west and south in the MAB; model results suggest substantial inter-population connectivity from southern New England to the Delmarva region. Simulated surfclam larvae generally drift for over one hundred kilometers along the shelf, but the distance traveled is highly variable in space and over time. Surfclam larval growth and transport are strongly impacted by the physical environment. This suggests the need to further examine how the interaction between environment, behavior, and physiology affects inter-population connectivity. Larval vertical swimming and sinking behaviors have a significant net effect of increasing larval drifting distances when compared with a purely passive model, confirming the need to include larval behavior.

  20. Cohesin, Gene Expression and Development: Lessons from Drosophila

    PubMed Central

    Dorsett, Dale

    2010-01-01

    The cohesin complex, discovered for its role in sister chromatid cohesion, also plays roles in gene expression and development in organisms from yeast to man. This review highlights what has been learned about the gene control and developmental functions of cohesin and the Nipped-B (NIPBL/Scc2) cohesin loading factor in Drosophila. The Drosophila studies have provided unique insights into the etiology of Cornelia de Lange syndrome (CdLS), which is caused by mutations affecting sister chromatid cohesion proteins in humans. In vivo experiments with Drosophila show that cohesin and Nipped-B have dosage-sensitive effects on the functions of many evolutionarily conserved genes and developmental pathways. Genome-wide studies with Drosophila cultured cells show that Nipped-B and cohesin co-localize on chromosomes, and bind preferentially, but not exclusively, to many actively-transcribed genes and their regulatory sequences, including many of the proposed in vivo target genes. In contrast, the cohesion factors are largely excluded from genes silenced by Polycomb group (PcG) proteins. Combined, the in vivo genetic data and the binding patterns of cohesin and Nipped-B in cultured cells are consistent with the hypothesis that they control the action of gene regulatory sequences, including transcriptional enhancers and insulators, and suggest that they might also help define active chromatin domains and influence transcriptional elongation. PMID:19308700

  1. Circulation constrains the evolution of larval development modes and life histories in the coastal ocean.

    PubMed

    Pringle, James M; Byers, James E; Pappalardo, Paula; Wares, John P; Marshall, Dustin

    2014-04-01

    The evolutionary pressures that drive long larval planktonic durations in some coastal marine organisms, while allowing direct development in others, have been vigorously debated. We introduce into the argument the asymmetric dispersal of larvae by coastal currents and find that the strength of the currents helps determine which dispersal strategies are evolutionarily stable. In a spatially and temporally uniform coastal ocean of finite extent, direct development is always evolutionarily stable. For passively drifting larvae, long planktonic durations are stable when the ratio of mean to fluctuating currents is small and the rate at which larvae increase in size in the plankton is greater than the mortality rate (both in units of per time). However, larval behavior that reduces downstream larval dispersal for a given time in plankton will be selected for, consistent with widespread observations of behaviors that reduce dispersal of marine larvae. Larvae with long planktonic durations are shown to be favored not for the additional dispersal they allow, but for the additional fecundity that larval feeding in the plankton enables. We analyzed the spatial distribution of larval life histories in a large database of coastal marine benthic invertebrates and documented a link between ocean circulation and the frequency of planktotrophy in the coastal ocean. The spatial variation in the frequency of species with planktotrophic larvae is largely consistent with our theory; increases in mean currents lead to a decrease in the fraction of species with planktotrophic larvae over a broad range of temperatures. PMID:24933820

  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. 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

  4. Exploring Larval Development and Applications in Marine Fish Aquaculture Using Pink Snapper Embryos

    ERIC Educational Resources Information Center

    Tamaru, Clyde; Haverkort-Yeh, Roxanne D.; Gorospe, Kelvin D.; Rivera, Malia Ana J.

    2014-01-01

    This biology investigation on "Pristipomoides filamentosus" larval development, survival, and aquaculture research was developed with three educational objectives: to provide high school students with (1) a scientific background on the biology and science of fisheries as well as overfishing, its consequences, and possible mitigations;…

  5. Worker honey bee ovary development: seasonal variation and the influence of larval and adult nutrition.

    PubMed

    Hoover, Shelley E R; Higo, Heather A; Winston, Mark L

    2006-01-01

    We examined the effect of larval and adult nutrition on worker honey bee (Apis mellifera L.) ovary development. Workers were fed high or low-pollen diets as larvae, and high or low-protein diets as adults. Workers fed low-protein diets at both life stages had the lowest levels of ovary development, followed by those fed high-protein diets as larvae and low- quality diets as adults, and then those fed diets poor in protein as larvae but high as adults. Workers fed high-protein diets at both life stages had the highest levels of ovary development. The increases in ovary development due to improved dietary protein in the larval and adult life stages were additive. Adult diet also had an effect on body mass. The results demonstrate that both carry-over of larval reserves and nutrients acquired in the adult life stage are important to ovary development in worker honey bees. Carry-over from larval development, however, appears to be less important to adult fecundity than is adult nutrition. Seasonal trends in worker ovary development and mass were examined throughout the brood rearing season. Worker ovary development was lowest in spring, highest in mid-summer, and intermediate in fall. PMID:16228242

  6. Building from the ground up: basement membranes in Drosophila development

    PubMed Central

    Isabella, Adam J.; Horne-Badovinac, Sally

    2016-01-01

    Basement Membranes (BMs) are sheet-like extracellular matrices found at the basal surfaces of epithelial tissues. The structural and functional diversity of these matrices within the body endows them with the ability to affect multiple aspects of cell behavior and communication; for this reason, BMs are integral to many developmental processes. The power of Drosophila genetics, as applied to the BM, has yielded substantial insight into how these matrices influence development. Here, we explore three facets of BM biology to which Drosophila research has made particularly important contributions. First we discuss how newly synthesized BM proteins are secreted to and assembled exclusively on basal epithelial surfaces. Next, we examine how regulation of the structural properties of the BM mechanically supports and guides tissue morphogenesis. Finally, we explore how BMs influence development through the modulation of several major signaling pathways. PMID:26610918

  7. 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

  8. Extracellular matrix and its receptors in Drosophila neural development

    PubMed Central

    Broadie, Kendal; Baumgartner, Stefan; Prokop, Andreas

    2011-01-01

    Extracellular matrix (ECM) and matrix receptors are intimately involved in most biological processes. The ECM plays fundamental developmental and physiological roles in health and disease, including processes underlying the development, maintenance and regeneration of the nervous system. To understand the principles of ECM-mediated functions in the nervous system, genetic model organisms like Drosophila provide simple, malleable and powerful experimental platforms. This article provides an overview of ECM proteins and receptors in Drosophila. It then focuses on their roles during three progressive phases of neural development: 1) neural progenitor proliferation, 2) axonal growth and pathfinding and 3) synapse formation and function. Each section highlights known ECM and ECM-receptor components and recent studies done in mutant conditions to reveal their in vivo functions, all illustrating the enormous opportunities provided when merging work on the nervous system with systematic research into ECM-related gene functions. PMID:21688401

  9. Genome-Wide Approaches to Drosophila Heart Development

    PubMed Central

    Frasch, Manfred

    2016-01-01

    The development of the dorsal vessel in Drosophila is one of the first systems in which key mechanisms regulating cardiogenesis have been defined in great detail at the genetic and molecular level. Due to evolutionary conservation, these findings have also provided major inputs into studies of cardiogenesis in vertebrates. Many of the major components that control Drosophila cardiogenesis were discovered based on candidate gene approaches and their functions were defined by employing the outstanding genetic tools and molecular techniques available in this system. More recently, approaches have been taken that aim to interrogate the entire genome in order to identify novel components and describe genomic features that are pertinent to the regulation of heart development. Apart from classical forward genetic screens, the availability of the thoroughly annotated Drosophila genome sequence made new genome-wide approaches possible, which include the generation of massive numbers of RNA interference (RNAi) reagents that were used in forward genetic screens, as well as studies of the transcriptomes and proteomes of the developing heart under normal and experimentally manipulated conditions. Moreover, genome-wide chromatin immunoprecipitation experiments have been performed with the aim to define the full set of genomic binding sites of the major cardiogenic transcription factors, their relevant target genes, and a more complete picture of the regulatory network that drives cardiogenesis. This review will give an overview on these genome-wide approaches to Drosophila heart development and on computational analyses of the obtained information that ultimately aim to provide a description of this process at the systems level. PMID:27294102

  10. Tissue-specific targeting of Hsp26 has no effect on heat resistance of neural function in larval Drosophila

    PubMed Central

    Mileva-Seitz, Viara; Xiao, Chengfeng; Seroude, Laurent

    2008-01-01

    Hsp26 belongs to the small heat-shock protein family and is normally expressed in all cells during heat stress. We aimed to determine if overexpression of this protein protects behavior and neural function in Drosophila melanogaster during heat stress, as has previously been shown for Hsp70. We used the UAS-GAL4 expression system to drive expression of Hsp26 in the whole animal (ubiquitously), in the motoneurons, and in the muscles of wandering third-instar larvae. There were slight increases in time to crawling failure and normalized excitatory junction potential (EJP) area for some of the transgenic lines, but these were not consistent. In addition, Hsp26 had no effect on the temperature at failure of EJPs, normalized EJP peak amplitude, and normalized EJP half-width. Overexpression larvae had a similar number of motoneuronal boutons and length of nerve terminals as controls, indicating that the occasional protective effects on locomotion were not due to changes at the synapse. We conclude that overexpression had a small thermoprotective effect on locomotion and no effect on neural function. As it has been shown that Hsp26 requires action of other Hsps to reactivate the denatured proteins to which it binds, we propose that at least in larvae, the function of Hsp26 was masked in the relative absence of other Hsps. PMID:18347945

  11. Genetic variation in locomotor activity rhythm among populations of Leptopilina heterotoma (Hymenoptera: Eucoilidae), a larval parasitoid of Drosophila species.

    PubMed

    Fleury, F; Allemand, R; Fouillet, P; Boulétreau, M

    1995-01-01

    The locomotor activity rhythm of Leptopilina heterotoma, a parasitoid insect of Drosophila larvae, was investigated under laboratory conditions. Under LD 12:12, the locomotor activity of females shows a clear rhythm which persists under continuous darkness (circadian rhythm). However, comparative study of five populations indicates that both the rate of activity and the profile of the rhythm vary according to the origin of females. The Mediterranean populations (Tunisia and Antibes) show two peaks of activity, at the beginning and at the end of the photophase, whereas more northern populations (Lyon and the Netherlands) are mostly active during the afternoon. Females originating from the area of Lyon have a very low level of activity. Reciprocal crosses (F1 hybrids and backcrosses) between the French and the Tunisian strains demonstrated the genetic basis of these variations and the biparental inheritance of the trait. This genetic variability is interpreted as a consequence of selective pressures and suggests a local adaptation of natural populations in host foraging behavior. The selective factors which could act on the daily organization of parasitoid behaviors are discussed. PMID:7755522

  12. Evolution and development of gastropod larval shell morphology: experimental evidence for mechanical defense and repair.

    PubMed

    Hickman, C S

    2001-01-01

    The structural diversity of gastropod veliger larvae offers an instructive counterpoint to the view of larval forms as conservative archetypes. Larval structure, function, and development are fine-tuned for survival in the plankton. Accordingly, the study of larval adaptation provides an important perspective for evolutionary-developmental biology as an integrated science. Patterns of breakage and repair in the field, as well as patterns of breakage in arranged encounters with zooplankton under laboratory conditions, are two powerful sources of data on the adaptive significance of morphological and microsculptural features of the gastropod larval shell. Shells of the planktonic veliger larvae of the caenogastropod Nassarius paupertus [GOULD] preserve multiple repaired breaks, attributed to unsuccessful zooplankton predators. In culture, larvae isolated from concentrated zooplankton samples rapidly repaired broken apertural margins and restored the "ideal" apertural form, in which an elaborate projection or "beak" covers the head of the swimming veliger. When individuals with repaired apertures were reintroduced to a concentrated mixture of potential zooplankton predators, the repaired margins were rapidly chipped and broken back. The projecting beak of the larval shell is the first line of mechanical defense, covering the larval head and mouth and potentially the most vulnerable part of the shell to breakage. Patterns of mechanical failure show that spiral ridges do reinforce the beak and retard breakage. The capacity for rapid shell repair and regeneration, and the evolution of features that resist or retard mechanical damage, may play a more prominent role than previously thought in enhancing the ability of larvae to survive in the plankton. PMID:11256430

  13. INFLUENCE OF AN INSECT GROWTH REGULATOR ON THE LARVAL DEVELOPMENT OF THE MUD CRAB RHITHOPANOPEUS HARRISSII

    EPA Science Inventory

    Effects of the juvenile hormone analogue (S)-methoprene on the larval development and survival of the estuarine mud crab hithropanopeus harrisii were examined in the laboratory. rablarvae continuously exposed to 1000 ug (S)-methoprene/liter did not survive beyond zoeal stage I. i...

  14. Development of a larval diet for the South American fruit fly Anastrepha fraterculus (Diptera:Tephritidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mass-rearing protocols must be developed. In particular, a cost-effective larval diet, to implement the sterile insect technique against Anastrepha fratercculus (Wiedemann). The key elements of this diet are the optimal nutrients and their concentrations, diet supports or bulking agents, and the pH ...

  15. Vacuole dynamics in the salivary glands of Drosophila melanogaster during prepupal development.

    PubMed

    Farkaš, Robert; Beňová-Liszeková, Denisa; Mentelová, Lucia; Mahmood, Silvia; Ďatková, Zuzana; Beňo, Milan; Pečeňová, Ludmila; Raška, Otakar; Šmigová, Jana; Chase, Bruce A; Raška, Ivan; Mechler, Bernard M

    2015-01-01

    A central function of the Drosophila salivary glands (SGs), historically known for their polytene chromosomes, is to produce and then release during pupariation the secretory glue used to affix a newly formed puparium to a substrate. This essential event in the life history of Drosophila is regulated by the steroid hormone ecdysone in the late-larval period. Ecdysone triggers a cascade of sequential gene activation that leads to glue secretion and initiates the developmentally-regulated programmed cell death (PCD) of the larval salivary glands, which culminates 16 h after puparium formation (APF). We demonstrate here that, even after the larval salivary glands have completed what is perceived to be one of their major biological functions--glue secretion during pupariation--they remain dynamic and physiologically active up until the execution phase of PCD. We have used specific metabolic inhibitors and genetic tools, including mutations or transgenes for shi, Rab5, Rab11, vha55, vha68-2, vha36-1, syx1A, syx4, and Vps35 to characterize the dramatic series of cellular changes occurring in the SG cells between pupariation and 7-8 h APF. Early in the prepupal period, they are remarkably active in endocytosis, forming acidic vacuoles. Midway through the prepupal period, there is abundant late endosomal trafficking and vacuole growth, which is followed later by vacuole neutralization and disappearance via membrane consolidation. This work provides new insights into the function of Drosophila SGs during the early- to mid-prepupal period. PMID:25611296

  16. 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

  17. 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

  18. Role of Bacillus thuringiensis Cry1 δ Endotoxin Binding in Determining Potency during Lepidopteran Larval Development

    PubMed Central

    Gilliland, Androulla; Chambers, Catherine E.; Bone, Eileen J.; Ellar, David J.

    2002-01-01

    Five economically important crop pests, Manduca sexta, Pieris brassicae, Mamestra brassicae, Spodoptera exigua, and Agrotis ipsilon, were tested at two stages of larval development for susceptibility to Bacillus thuringiensis toxins Cry1Ac, Cry1Ca, Cry1J, and Cry1Ba. Bioassay results for M. sexta showed that resistance to all four Cry toxins increased from the neonate stage to the third-instar stage; the increase in resistance was most dramatic for Cry1Ac, the potency of which decreased 37-fold. More subtle increases in resistance during larval development were seen in M. brassicae for Cry1Ca and in P. brassicae for Cry1Ac and Cry1J. By contrast, the sensitivity of S. exigua did not change during development. At both larval stages, A. ipsilon was resistant to all four toxins. Because aminopeptidase N (APN) is a putative Cry1 toxin binding protein, APN activity was measured in neonate and third-instar brush border membrane vesicles (BBMV). With the exception of S. exigua, APN activity was found to be significantly lower in neonates than in third-instar larvae and thus inversely correlated with increased resistance during larval development. The binding characteristics of iodinated Cry1 toxins were determined for neonate and third-instar BBMV. In M. sexta, the increased resistance to Cry1Ac and Cry1Ba during larval development was positively correlated with fewer binding sites in third-instar BBMV than in neonate BBMV. The other species-instar-toxin combinations did not reveal positive correlations between potency and binding characteristics. The correlation between binding and potency was inconsistent for the species-instar-toxin combinations used in this study, reaffirming the complex mode of action of Cry1 toxins. PMID:11916662

  19. Development of the Acoustically Evoked Behavioral Response in Larval Plainfin Midshipman Fish, Porichthys notatus

    PubMed Central

    Alderks, Peter W.; Sisneros, Joseph A.

    2013-01-01

    The ontogeny of hearing in fishes has become a major interest among bioacoustics researchers studying fish behavior and sensory ecology. Most fish begin to detect acoustic stimuli during the larval stage which can be important for navigation, predator avoidance and settlement, however relatively little is known about the hearing capabilities of larval fishes. We characterized the acoustically evoked behavioral response (AEBR) in the plainfin midshipman fish, Porichthys notatus, and used this innate startle-like response to characterize this species' auditory capability during larval development. Age and size of larval midshipman were highly correlated (r2 = 0.92). The AEBR was first observed in larvae at 1.4 cm TL. At a size ≥1.8 cm TL, all larvae responded to a broadband stimulus of 154 dB re1 µPa or −15.2 dB re 1 g (z-axis). Lowest AEBR thresholds were 140–150 dB re 1 µPa or −33 to −23 dB re 1 g for frequencies below 225 Hz. Larval fish with size ranges of 1.9–2.4 cm TL had significantly lower best evoked frequencies than the other tested size groups. We also investigated the development of the lateral line organ and its function in mediating the AEBR. The lateral line organ is likely involved in mediating the AEBR but not necessary to evoke the startle-like response. The midshipman auditory and lateral line systems are functional during early development when the larvae are in the nest and the auditory system appears to have similar tuning characteristics throughout all life history stages. PMID:24340003

  20. Cell position during larval development affects postdiapause development in Megachile rotundata (Hymenoptera: Megachilidae).

    PubMed

    Yocum, George D; Rinehart, Joseph P; Kemp, William P

    2014-08-01

    Megachile rotundata (F.) (Hymenoptera: Megachilidae) is the primary pollinator of alfalfa in the northwestern United States and western Canada and provides pollination services for onion, carrot, hybrid canola, various legumes, and other specialty crops. M. rotundata females are gregarious, nest in cavities either naturally occurring or in artificial nesting blocks, where they construct a linear series of brood cells. Because of the physical layout of the nest, the age of the larvae within the nest and the microenvironment the individual larvae experience will vary. These interacting factors along with other maternal inputs affect the resulting phenotypes of the nest mates. To further our understanding of in-nest physiology, gender and developmental rates were examined in relationship to cell position within the nest. Eighty-two percent of the females were located within the first three cells, those furthest from the nest entrance. For those individuals developing in cells located in the deepest half of the nest, the sex of the previous bee had a significant effect on the female decision of the gender of the following nest mate. Removing the prepupae from the nest and rearing them under identical conditions demonstrated that position within the nest during larval development had a significant effect on the postdiapause developmental rates, with males whose larval development occurred deeper in the nest developing more slowly than those toward the entrance. No positional effect on postdiapause developmental rates was noted for the females. The cell position effect on male postdiapause developmental rate demonstrates that postdiapause development is not a rigid physiological mechanism uniform in all individuals, but is a dynamic plastic process shaped by past environmental conditions. PMID:24914676

  1. Larval development and metamorphosis in Pleurobranchaea maculata, with a review of development in the notaspidea (Opisthobranchia).

    PubMed

    Gibson, Glenys D

    2003-10-01

    Pleurobranchaea maculata is a carnivorous notaspidean that is common in New Zealand. This species produces small eggs (diameter 100 microm) and planktotrophic veligers that hatch in 8 d and are planktonic for 3 weeks before settling on biofilmed surfaces (14 degrees C). Larval development is known in detail for only two other notaspidean species, P. japonica and Berthellina citrina. In all three species of pleurobranchids, mantle and shell growth show striking differences from veligers of other opisthobranch taxa. In young veligers of pleurobranchids, the shell is overgrown by the mantle, new shell is added by cells other than those of the mantle fold, and an operculum does not form. Thus some "adult" traits (e.g., notum differentiation, mechanism of shell growth, lack of operculum) are expressed early in larval development. This suggests that apomorphies characteristic of adult pleurobranchids evolved through heterochrony, with expression in larvae of traits typical of adults of other clades. The protoconch is dissolved post-settlement and not cast off as occurs in other opisthobranch orders, indicating that shell loss is apomorphic. P. maculata veligers are atypical of opisthobranchs in having a field of highly folded cells on the lower velar surface, a mouth that is posterior to the metatroch, and a richly glandular, possibly chemodefensive mantle. These data indicate that notaspidean larvae are highly derived in terms of the novel traits and the timing of morphogenic events. Phylogenetic analysis must consider embryological origins before assuming homology, as morphological similarities (e.g., shell loss) may have developed through distinct mechanisms. PMID:14583510

  2. Warmer winters reduce frog fecundity and shift breeding phenology, which consequently alters larval development and metamorphic timing.

    PubMed

    Benard, Michael F

    2015-03-01

    One widely documented phenological response to climate change is the earlier occurrence of spring-breeding events. While such climate change-driven shifts in phenology are common, their consequences for individuals and populations have rarely been investigated. I addressed this gap in our knowledge by using a multi-year observational study of six wood frog (Rana sylvatica) populations near the southern edge of their range. I tested first if winter temperature or precipitation affected the date of breeding and female fecundity, and second if timing of breeding affected subsequent larval development rate, mass at metamorphosis, date of metamorphosis, and survival. Warmer winters were associated with earlier breeding but reduced female fecundity. Winter precipitation did not affect breeding date, but was positively associated with female fecundity. There was no association between earlier breeding and larval survival or mass at metamorphosis, but earlier breeding was associated with delayed larval development. The delay in larval development was explained through a counterintuitive correlation between breeding date and temperature during larval development. Warmer winters led to earlier breeding, which in turn was associated with cooler post-breeding temperatures that slowed larval development. The delay in larval development did not fully compensate for the earlier breeding, such that for every 2 days earlier that breeding took place, the average date of metamorphosis was 1 day earlier. Other studies have found that earlier metamorphosis is associated with increased postmetamorphic growth and survival, suggesting that earlier breeding has beneficial effects on wood frog populations. PMID:25263760

  3. Energy metabolism during larval development of green and white abalone, Haliotis fulgens and H. sorenseni.

    PubMed

    Moran, Amy L; Manahan, Donal T

    2003-06-01

    An understanding of the biochemical and physiological energetics of lecithotrophic development is useful for interpreting patterns of larval development, dispersal potential, and life-history evolution. This study investigated the metabolic rates and use of biochemical reserves in two species of abalone, Haliotis fulgens (the green abalone) and H. sorenseni (the white abalone). Larvae of H. fulgens utilized triacylglycerol as a primary source of endogenous energy reserves for development ( approximately 50% depletion from egg to metamorphic competence). Amounts of phospholipid remained constant, and protein dropped by about 30%. After embryogenesis, larvae of H. fulgens had oxygen consumption rates of 81.7 +/- 5.9 (SE) pmol larva(-1) h(-1) at 15 degrees C through subsequent development. The loss of biochemical reserves fully met the needs of metabolism, as measured by oxygen consumption. Larvae of H. sorenseni were examined during later larval development and were metabolically and biochemically similar to H. fulgens larvae at a comparable stage. Metabolic rates of both species were very similar to previous data for a congener, H. rufescens, suggesting that larval metabolism and energy utilization may be conserved among closely related species that also share similar developmental morphology and feeding modes. PMID:12807704

  4. Influence of resources on Hermetia illucens (Diptera: Stratiomyidae) larval development.

    PubMed

    Nguyen, Trinh T X; Tomberlin, Jeffery K; Vanlaerhoven, Sherah

    2013-07-01

    Arthropod development can be used to determine the time of colonization of human remains to infer a minimum postmortem interval. The black soldier fly, Hermetia illucens L. (Diptera. Stratiomyidae) is native to North America and is unique in that its larvae can consume a wide range of decomposing organic material, including carrion. Larvae development was observed on six resources: control poultry feed, liver, manure, kitchen waste, fruits and vegetables, and fish rendering. Larvae fed manure were shorter, weighed less, and took longer to develop. Kitchen waste produced longer and heavier larvae, whereas larvae fed fish had almost 100% mortality. Black soldier flies can colonize human remains, which in many instances can coincide with food and organic wastes. Therefore, it is necessary to understand black soldier fly development on different food resources other than carrion tissue to properly estimate their age when recovered from human remains. PMID:23926790

  5. Development of a two photon microscope for tracking Drosophila larvae

    NASA Astrophysics Data System (ADS)

    Karagyozov, Doycho; Mihovilovic Skanata, Mirna; Gershow, Marc

    Current in vivo methods for measuring neural activity in Drosophila larva require immobilization of the animal. Although we can record neural signals while stimulating the sensory organs, we cannot read the behavioral output because we have prevented the animal from moving. Many research questions cannot be answered without observation of neural activity in behaving (freely-moving) animals. Our project aims to develop a tracking microscope that maintains the neurons of interest in the field of view and in focus during the rapid three dimensional motion of a free larva.

  6. Larval Development of Two N. E. Pacific Pilidiophoran Nemerteans (Heteronemertea; Lineidae).

    PubMed

    Hiebert, Terra C; Maslakova, Svetlana A

    2015-12-01

    Unique to the phylum Nemertea, the pilidium is an unmistakable planktonic larva found in one group of nemerteans, the Pilidiophora. Inside the pilidium, the juvenile develops from a series of epidermal invaginations in the larval body, called imaginal discs. The discs grow and fuse around the larval gut over the course of weeks to months in the plankton. Once complete, the juvenile breaks free from the larval body in a catastrophic metamorphosis, and often devours the larva as its first meal. One third of nemertean species are expected to produce a pilidium, but the larvae are known for very few species; development from fertilization to metamorphosis has been described in only one species, Micrura alaskensis. Known pilidia include both planktotrophic and lecithotrophic forms, and otherwise exhibit great morphological diversity. Here, we describe the complete development in two lineiform species that are common to the northeast Pacific coast, Micrura wilsoni and Lineus sp. "red." Both species possess typical, cap-shaped planktotrophic pilidia, and the order of emergence of imaginal discs is similar to that which is described in M. alaskensis. The pilidium of Lineus sp. "red" resembles pilidia of several other species, such as Lineus flavescens, and potentially characterizes a pilidiophoran clade. M. wilsoni has relatively transparent oocytes and a pilidium with what appears to be a unique pattern of pigmentation. The adults of both species are more commonly observed in intertidal zones than their larvae are in the plankton. PMID:26695825

  7. Imaging neural development in embryonic and larval sea urchins.

    PubMed

    Krupke, Oliver; Yaguchi, Shunsuke; Yaguchi, Junko; Burke, Robert D

    2014-01-01

    Imaging is a critical tool in neuroscience, and our understanding of the structure and function of sea urchin nervous systems owes much to this approach. In particular, studies of neural development have been facilitated by methods that enable the accurate identification of specific types of neurons. Here we describe methods that have been successfully employed to study neural development in sea urchin embryos. Altering gene expression in part of an embryo is facilitated by injection of reagents into individual blastomeres, which enables studies of cell autonomous effects and single embryo rescue experiments. The simultaneous localization of an in situ RNA hybridization probe and a cell type specific antigen has enabled studies of gene expression in specific types of neurons. Fixatives and antibodies can be capricious; thus, we provide data on preservation of antigens with commonly used fixatives and buffers. PMID:24567212

  8. Individual and mixture effects of selected pharmaceuticals on larval development of the estuarine shrimp Palaemon longirostris.

    PubMed

    González-Ortegón, Enrique; Blasco, Julian; Nieto, Elena; Hampel, Miriam; Le Vay, Lewis; Giménez, Luis

    2016-01-01

    Few ecotoxicological studies incorporate within the experimental design environmental variability and mixture effects when assessing the impact of pollutants on organisms. We have studied the combined effects of selected pharmaceutical compounds and environmental variability in terms of salinity and temperature on survival, development and body mass of larvae of the estuarine shrimp Palaemon longirostris. Drug residues found in coastal waters occur as mixture, and the evaluation of combined effects of simultaneously occurring compounds is indispensable for their environmental risk assessment. All larval stages of P. longirostris were exposed to the nonsteroidal anti-inflammatory drug (NSAID) diclofenac sodium (DS: 40 and 750 μg L(-1)), the lipid regulator clofibric acid (CA: 17 and 361 μg L(-1)) and the fungicide clotrimazole (CLZ: 0.14 and 4 μg L(-1)). We observed no effect on larval survival of P. longirostris with the tested pharmaceuticals. However, and in contrast to previous studies on larvae of the related marine species Palaemon serratus, CA affected development through an increase in intermoult duration and reduced growth without affecting larval body mass. These developmental effects in P. longirostris larvae were similar to those observed in the mixture of DS and CA confirming the toxic effects of CA. In the case of CLZ, its effects were similar to those observed previously in P. serratus: high doses affected development altering intermoult duration, tended to reduce the number of larval instars and decreased significantly the growth rate. This study suggests that an inter-specific life histories approach should be taken into account to assess the effect of emergent compounds in coastal waters. PMID:26163379

  9. Using MARCM to study Drosophila brain development.

    PubMed

    Viktorin, Gudrun

    2014-01-01

    Mosaic analysis with a repressible cell marker (MARCM) generates positively labeled, wild-type or mutant mitotic clones by unequally distributing a repressor of a cell lineage marker, originally tubP-driven GAL80 repressing the GAL4/UAS system. Variations of the technique include labeling of both sister clones (twin spot MARCM), the simultaneous use of two different drivers within the same clone (dual MARCM), as well as the use of different repressible transcription systems (Q-MARCM). MARCM can be combined with any UAS-based construct, such as localized GFP fusions to visualize subcellular compartments, genes for rescue and ectopic expression, and modifiers of neural activity. A related technique, the twin spot generator, generates positively labeled clones without the use of a repressor, thus minimizing the lag time between clone induction and appearance of label. The present protocol provides a detailed description of a standard MARCM analysis of brain development that includes generation of MARCM stocks and crosses, induction of clones, brain dissection at various stages of development, immunohistochemistry, and confocal microscopy, and can be modified for similar experiments involving mitotic clones. PMID:24048928

  10. The silkworm glutathione S-transferase gene noppera-bo is required for ecdysteroid biosynthesis and larval development.

    PubMed

    Enya, Sora; Daimon, Takaaki; Igarashi, Fumihiko; Kataoka, Hiroshi; Uchibori, Miwa; Sezutsu, Hideki; Shinoda, Tetsuro; Niwa, Ryusuke

    2015-06-01

    Insect molting and metamorphosis are tightly controlled by ecdysteroids, which are important steroid hormones that are synthesized from dietary sterols in the prothoracic gland. One of the ecdysteroidogenic genes in the fruit fly Drosophila melanogaster is noppera-bo (nobo), also known as GSTe14, which encodes a member of the epsilon class of glutathione S-transferases. In D. melanogaster, nobo plays a crucial role in utilizing cholesterol via regulating its transport and/or metabolism in the prothoracic gland. However, it is still not known whether the orthologs of nobo from other insects are also involved in ecdysteroid biosynthesis via cholesterol transport and/or metabolism in the prothoracic gland. Here we report genetic evidence showing that the silkworm Bombyx mori ortholog of nobo (nobo-Bm; GSTe7) is essential for silkworm development. nobo-Bm is predominantly expressed in the prothoracic gland. To assess the functional importance of nobo-Bm, we generated a B. mori genetic mutant of nobo-Bm using TALEN-mediated genome editing. We show that loss of nobo-Bm function causes larval arrest and a glossy cuticle phenotype, which are rescued by the application of 20-hydroxyecdysone. Moreover, the prothoracic gland cells isolated from the nobo-Bm mutant exhibit an abnormal accumulation of 7-dehydrocholesterol, a cholesterol metabolite. These results suggest that the nobo family of glutathione S-transferases is essential for development and for the regulation of sterol utilization in the prothoracic gland in not only the Diptera but also the Lepidoptera. On the other hand, loss of nobo function mutants of D. melanogaster and B. mori abnormally accumulates different sterols, implying that the sterol utilization in the PG is somewhat different between these two insect species. PMID:25881968

  11. UV wavelengths experienced during development affect larval newt visual sensitivity and predation efficiency.

    PubMed

    Martin, Mélissa; Théry, Marc; Rodgers, Gwendolen; Goven, Delphine; Sourice, Stéphane; Mège, Pascal; Secondi, Jean

    2016-02-01

    We experimentally investigated the influence of developmental plasticity of ultraviolet (UV) visual sensitivity on predation efficiency of the larval smooth newt, Lissotriton vulgaris. We quantified expression of SWS1 opsin gene (UV-sensitive protein of photoreceptor cells) in the retinas of individuals who had developed in the presence (UV+) or absence (UV-) of UV light (developmental treatments), and tested their predation efficiency under UV+ and UV- light (testing treatments). We found that both SWS1 opsin expression and predation efficiency were significantly reduced in the UV- developmental group. Larvae in the UV- testing environment displayed consistently lower predation efficiency regardless of their developmental treatment. These results prove for the first time, we believe, functional UV vision and developmental plasticity of UV sensitivity in an amphibian at the larval stage. They also demonstrate that UV wavelengths enhance predation efficiency and suggest that the magnitude of the behavioural response depends on retinal properties induced by the developmental lighting environment. PMID:26843556

  12. Postembryonic lineages of the Drosophila brain: I. Development of the lineage-associated fiber tracts

    PubMed Central

    Lovick, Jennifer K.; Ngo, Kathy T.; Omoto, Jaison J.; Wong, Darren C.; Nguyen, Joseph D.; Hartenstein, Volker

    2013-01-01

    Neurons of the Drosophila central brain fall into approximately 100 paired groups, termed lineages. Each lineage is derived from a single asymmetrically-dividing neuroblast. Embryonic neuroblasts produce 1,500 primary neurons (per hemisphere) that make up the larval CNS followed by a second mitotic period in the larva that generates approximately 10,000 secondary, adult-specific neurons. Clonal analyses based on previous works using lineage-specific Gal4 drivers have established that such lineages form highly invariant morphological units. All neurons of a lineage project as one or a few axon tracts (secondary axon tracts, SATs) with characteristic trajectories, thereby representing unique hallmarks. In the neuropil, SATs assemble into larger fiber bundles (fascicles) which interconnect different neuropil compartments. We have analyzed the SATs and fascicles formed by lineages during larval, pupal, and adult stages using antibodies against membrane molecules (Neurotactin/Neuroglian) and synaptic proteins (Bruchpilot/N-Cadherin). The use of these markers allows one to identify fiber bundles of the adult brain and associate them with SATs and fascicles of the larval brain. This work lays the foundation for assigning the lineage identity of GFP-labeled MARCM clones on the basis of their close association with specific SATs and neuropil fascicles, as described in the accompanying paper (Wong et al., 2013. Postembryonic lineages of the Drosophila brain: II. Identification of lineage projection patterns based on MARCM clones. Submitted.). PMID:23880429

  13. Postembryonic lineages of the Drosophila brain: I. Development of the lineage-associated fiber tracts.

    PubMed

    Lovick, Jennifer K; Ngo, Kathy T; Omoto, Jaison J; Wong, Darren C; Nguyen, Joseph D; Hartenstein, Volker

    2013-12-15

    Neurons of the Drosophila central brain fall into approximately 100 paired groups, termed lineages. Each lineage is derived from a single asymmetrically-dividing neuroblast. Embryonic neuroblasts produce 1,500 primary neurons (per hemisphere) that make up the larval CNS followed by a second mitotic period in the larva that generates approximately 10,000 secondary, adult-specific neurons. Clonal analyses based on previous works using lineage-specific Gal4 drivers have established that such lineages form highly invariant morphological units. All neurons of a lineage project as one or a few axon tracts (secondary axon tracts, SATs) with characteristic trajectories, thereby representing unique hallmarks. In the neuropil, SATs assemble into larger fiber bundles (fascicles) which interconnect different neuropil compartments. We have analyzed the SATs and fascicles formed by lineages during larval, pupal, and adult stages using antibodies against membrane molecules (Neurotactin/Neuroglian) and synaptic proteins (Bruchpilot/N-Cadherin). The use of these markers allows one to identify fiber bundles of the adult brain and associate them with SATs and fascicles of the larval brain. This work lays the foundation for assigning the lineage identity of GFP-labeled MARCM clones on the basis of their close association with specific SATs and neuropil fascicles, as described in the accompanying paper (Wong et al., 2013. Postembryonic lineages of the Drosophila brain: II. Identification of lineage projection patterns based on MARCM clones. Submitted.). PMID:23880429

  14. Spawning, fertilization, and larval development of Potamocorbula amurensis (Mollusca: Bivalvia) from San Francisco Bay, California

    USGS Publications Warehouse

    Nicolini, M.H.; Penry, D.L.

    2000-01-01

    In Potamocorbula amurensis time for development to the straight-hinge larval stage is 48 hr at 15??C. Potamocorbula amurensis settles at a shell length of approximately 135 ??m 17 to 19 days after fertilization. Our observations of timing of larval devdlopment in P. amurensis support the hypothesis of earlier workers that its route of initial introduction to San Francisco Bay was as veliger larvae transported in ballast water by trans-Pacific cargo ships. The length of the larval period of P. amurensis relative to water mass residence times in San Francisco Bay suggests that it is sufficient to allow substantial dispersal from North Bay to South Bay populations in concordance with previous observations that genetic differentiation among populations of P. amurensis in San Francisco Bay is low. Potamocorbula amurensis is markedly euryhaline at all stages of development. Spawning and fertilization can occur at salinities from 5 to 25 psu, and eggs and sperms can each tolerance at least a 10-psu step increase or decrease in salinity. Embryos that are 2 hr old can tolerate the same range of salinities from (10 to 30 psu), and by the time they are 24 hr old they can tolerate the same range of salinities (2 to 30 psu) that adult clams can. The ability of P. amurensis larvae to tolerate substantial step changes in salinity suggests a strong potential to survive incomplete oceanic exchanges of ballast water and subsequent discharge into receiving waters across a broad range of salinities.

  15. Ferritin Is Required in Multiple Tissues during Drosophila melanogaster Development

    PubMed Central

    Blowes, Liisa M.; Missirlis, Fanis; Riesgo-Escovar, Juan R.

    2015-01-01

    In Drosophila melanogaster, iron is stored in the cellular endomembrane system inside a protein cage formed by 24 ferritin subunits of two types (Fer1HCH and Fer2LCH) in a 1:1 stoichiometry. In larvae, ferritin accumulates in the midgut, hemolymph, garland, pericardial cells and in the nervous system. Here we present analyses of embryonic phenotypes for mutations in Fer1HCH, Fer2LCH and in both genes simultaneously. Mutations in either gene or deletion of both genes results in a similar set of cuticular embryonic phenotypes, ranging from non-deposition of cuticle to defects associated with germ band retraction, dorsal closure and head involution. A fraction of ferritin mutants have embryonic nervous systems with ventral nerve cord disruptions, misguided axonal projections and brain malformations. Ferritin mutants die with ectopic apoptotic events. Furthermore, we show that ferritin maternal contribution, which varies reflecting the mother’s iron stores, is used in early development. We also evaluated phenotypes arising from the blockage of COPII transport from the endoplasmic reticulum to the Golgi apparatus, feeding the secretory pathway, plus analysis of ectopically expressed and fluorescently marked Fer1HCH and Fer2LCH. Overall, our results are consistent with insect ferritin combining three functions: iron storage, intercellular iron transport, and protection from iron-induced oxidative stress. These functions are required in multiple tissues during Drosophila embryonic development. PMID:26192321

  16. Genetic control of epithelial tube fusion during Drosophila tracheal development.

    PubMed

    Samakovlis, C; Manning, G; Steneberg, P; Hacohen, N; Cantera, R; Krasnow, M A

    1996-11-01

    During development of tubular networks such as the mammalian vascular system, the kidney and the Drosophila tracheal system, epithelial tubes must fuse to each other to form a continuous network. Little is known of the cellular mechanisms or molecular control of epithelial tube fusion. We describe the cellular dynamics of a tracheal fusion event in Drosophila and identify a gene regulatory hierarchy that controls this extraordinary process. A tracheal cell located at the developing fusion point expresses a sequence of specific markers as it grows out and contacts a similar cell from another tube; the two cells adhere and form an intercellular junction, and they become doughnut-shaped cells with the lumen passing through them. The early fusion marker Fusion-1 is identified as the escargot gene. It lies near the top of the regulatory hierarchy, activating the expression of later fusion markers and repressing genes that promote branching. Ectopic expression of escargot activates the fusion process and suppresses branching throughout the tracheal system, leading to ectopic tracheal connections that resemble certain arteriovenous malformations in humans. This establishes a simple genetic system to study fusion of epithelial tubes. PMID:8951068

  17. Neuroblast lineage identification and lineage-specific Hox gene action during postembryonic development of the subesophageal ganglion in the Drosophila central brain.

    PubMed

    Kuert, Philipp A; Hartenstein, Volker; Bello, Bruno C; Lovick, Jennifer K; Reichert, Heinrich

    2014-06-15

    The central brain of Drosophila consists of the supraesophageal ganglion (SPG) and the subesophageal ganglion (SEG), both of which are generated by neural stem cell-like neuroblasts during embryonic and postembryonic development. Considerable information has been obtained on postembryonic development of the neuroblasts and their lineages in the SPG. In contrast, very little is known about neuroblasts, neural lineages, or any other aspect of the postembryonic development in the SEG. Here we characterize the neuroanatomy of the larval SEG in terms of tracts, commissures, and other landmark features as compared to a thoracic ganglion. We then use clonal MARCM labeling to identify all adult-specific neuroblast lineages in the late larval SEG and find a surprisingly small number of neuroblast lineages, 13 paired and one unpaired. The Hox genes Dfd, Scr, and Antp are expressed in a lineage-specific manner in these lineages during postembryonic development. Hox gene loss-of-function causes lineage-specific defects in axonal targeting and reduction in neural cell numbers. Moreover, it results in the formation of novel ectopic neuroblast lineages. Apoptosis block also results in ectopic lineages suggesting that Hox genes are required for lineage-specific termination of proliferation through programmed cell death. Taken together, our findings show that postembryonic development in the SEG is mediated by a surprisingly small set of identified lineages and requires lineage-specific Hox gene action to ensure the correct formation of adult-specific neurons in the Drosophila brain. PMID:24713419

  18. Genetic analysis of the bone morphogenetic protein-related gene, gbb, identifies multiple requirements during Drosophila development.

    PubMed Central

    Wharton, K A; Cook, J M; Torres-Schumann, S; de Castro, K; Borod, E; Phillips, D A

    1999-01-01

    We have isolated mutations in the Drosophila melanogaster gene glass bottom boat (gbb), which encodes a TGF-beta signaling molecule (formerly referred to as 60A) with highest sequence similarity to members of the bone morphogenetic protein (BMP) subgroup including vertebrate BMPs 5-8. Genetic analysis of both null and hypomorphic gbb alleles indicates that the gene is required in many developmental processes, including embryonic midgut morphogenesis, patterning of the larval cuticle, fat body morphology, and development and patterning of the imaginal discs. In the embryonic midgut, we show that gbb is required for the formation of the anterior constriction and for maintenance of the homeotic gene Antennapedia in the visceral mesoderm. In addition, we show a requirement for gbb in the anterior and posterior cells of the underlying endoderm and in the formation and extension of the gastric caecae. gbb is required in all the imaginal discs for proper disc growth and for specification of veins in the wing and of macrochaete in the notum. Significantly, some of these tissues have been shown to also require the Drosophila BMP2/4 homolog decapentaplegic (dpp), while others do not. These results indicate that signaling by both gbb and dpp may contribute to the development of some tissues, while in others, gbb may signal independently of dpp. PMID:10353905

  19. Juvenile Green Frog (Rana clamitans) Predatory Ability not Affected by Exposure to Carbaryl at Different Times During Larval Development

    PubMed Central

    Davis, Melanie J.; Kleinhenz, Peter; Boone, Michelle D.

    2011-01-01

    Larval exposure to pesticides can occur at different times during development, and can negatively impact amphibian fitness. We examined the effects of larval green frog (Rana clamitans) exposure to carbaryl at 2, 4, 8, or 16 weeks of development on juvenile predatory ability. We did not find evidence that predatory ability was affected by exposure to carbaryl, which suggests that carbaryl does not have latent effects on the predatory performance of green frogs in subsequent life stages. PMID:21462236

  20. Cancer Drug Development Using Drosophila as an in vivo Tool: From Bedside to Bench and Back.

    PubMed

    Yadav, Amarish Kumar; Srikrishna, Saripella; Gupta, Subash Chandra

    2016-09-01

    The fruit fly Drosophila melanogaster has been used for modeling cancer and as an in vivo tool for the validation and/or development of cancer therapeutics. The impetus for the use of Drosophila in cancer research stems from the high conservation of its signaling pathways, lower genetic redundancy, short life cycle, genetic amenability, and ease of maintenance. Several cell signaling pathways in Drosophila have been used for cancer drug development. The efficacy of combination therapy and uptake/bioavailability of drugs have also been studied. Drosophila has been validated using several FDA-approved drugs, suggesting a potential application of this model in drug repurposing. The model is emerging as a powerful tool for high-throughput screening and should significantly reduce the cost and time associated with drug development. In this review we discuss the applications of Drosophila in cancer drug development. The advantages and limitations of the model are discussed. PMID:27298020

  1. CCDC-55 is required for larval development and distal tip cell migration in Caenorhabditis elegans.

    PubMed

    Kovacevic, Ismar; Ho, Richard; Cram, Erin J

    2012-01-01

    The Caenorhabditis elegans distal tip cells (DTCs) are an in vivo model for the study of developmentally regulated cell migration. In this study, we characterize a novel role for CCDC-55, a conserved coiled-coil domain containing protein, in DTC migration and larval development in C. elegans. Although animals homozygous for a probable null allele, ccdc-55(ok2851), display an early larval arrest, RNAi depletion experiments allow the analysis of later phenotypes and suggest that CCDC-55 is needed within the DTC for migration to cease at the end of larval morphogenesis. The ccdc-55 gene is found in an operon with rnf-121 and rnf-5, E3 ubiquitin ligases that target cell migration genes such as the β-integrin PAT-3. Genetic interaction studies using RNAi depletion and the deletion alleles rnf-121(ok848) and rnf-5(tm794) indicate that CCDC-55 and the RNF genes act at least partially in parallel to promote termination of cell migration in the adult DTC. PMID:22285439

  2. CCDC-55 is required for larval development and distal tip cell migration in C. elegans

    PubMed Central

    Kovacevic, Ismar; Ho, Richard; Cram, Erin J.

    2012-01-01

    The C. elegans distal tip cells (DTCs) are an in vivo model for the study of developmentally regulated cell migration. In this study we characterize a novel role for CCDC-55, a conserved coiled-coil domain containing protein, in DTC migration and larval development in C. elegans. Although animals homozygous for a probable null allele, ccdc-55(ok2851), display an early larval arrest, RNAi depletion experiments allow the analysis of later phenotypes and suggest that CCDC-55 is needed within the DTC for migration to cease at the end of larval morphogenesis. The ccdc-55 gene is found in an operon with rnf-121 and rnf-5, E3 ubiquitin ligases that target cell migration genes such as the β-integrin PAT-3. Genetic interaction studies using RNAi depletion and the deletion alleles rnf-121(ok848) and rnf-5(tm794) indicate that CCDC-55 and the RNF genes act at least partially in parallel to promote termination of cell migration in the adult DTC. PMID:22285439

  3. Artificial substrates for oviposition and larval development of the pepper weevil (Coleoptera: Curculionidae).

    PubMed

    Addesso, K M; McAuslane, H J; Stansly, P A; Slansky, F; Schuster, D J

    2009-02-01

    The pepper weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae), is a major pest of cultivated peppers (Capsicum spp.) and other cultivated and wild species within the family Solanaceae. Laboratory study of this insect, as well as its biological control agents, will be greatly facilitated by an artificial rearing system that does not rely on pepper fruit. An egg collection method and amendments to a standard larval diet were investigated for use in the rearing of this weevil. Spherical sachets made of Parafilm or netting enclosing leaves of pepper, American black nightshade, eggplant, tomato, potato, and jasmine tobacco induced oviposition. Tomato, potato, and jasmine tobacco leaves were accepted despite the fact that these are not oviposition hosts for pepper weevils in the wild. A standard larval diet formula was modified in an attempt to improve egg hatch, larval survival, developmental time, and adult mass. The diet formula was modified with the addition of freeze-dried jalapeño pepper powder, an additional lipid source, alternate protein sources, and the removal of methyl paraben. None of the aforementioned treatments resulted in a significant improvement over the standard diet. Egg hatch was greater when eggs were incubated on moist paper towels rather than in diet; thus, placement of neonates rather than eggs into diet improved production of adults. Suggestions for more efficient rearing of weevils on the currently available diet and future directions for the development of an artificial rearing system for pepper weevil are discussed. PMID:19253644

  4. Regulation of twin of eyeless during Drosophila development.

    PubMed

    Skottheim Honn, John; Johansson, Linn; Rasmuson Lestander, Åsa

    2016-03-01

    The Pax-6 protein is vital for eye development in all seeing animals, from sea urchins to humans. Either of the Pax6 genes in Drosophila (twin of eyeless and eyeless) can induce a gene cascade leading to formation of entire eyes when expressed ectopically. The twin of eyeless (toy) gene in Drosophila is expressed in the anterior region of the early fly embryo. At later stages it is expressed in the brain, ventral nerve cord and (eventually) the visual primordium that gives rise to the eye-antennal imaginal discs of the larvae. These discs subsequently form the major part of the adult head, including compound eyes. We have searched for genes that are required for normal toy expression in the early embryo to elucidate initiating events of eye organogenesis. Candidate genes identified by mutation analyses were subjected to further knock-out and miss-expression tests to investigate their interactions with toy. Our results indicate that the head-specific gap gene empty spiracles can act as a repressor of Toy, while ocelliless (oc) and spalt major (salm) appear to act as positive regulators of toy gene expression. PMID:26976323

  5. Biotic and abiotic factors impacting development, behavior, phenology, and reproductive biology 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...

  6. Expression and light-triggered movement of rhodopsins in the larval visual system of mosquitoes

    PubMed Central

    Rocha, Manuel; Kimler, Kyle J.; Leming, Matthew T.; Hu, Xiaobang; Whaley, Michelle A.; O'Tousa, Joseph E.

    2015-01-01

    ABSTRACT During the larval stages, the visual system of the mosquito Aedes aegypti contains five stemmata, often referred to as larval ocelli, positioned laterally on each side of the larval head. Here we show that stemmata contain two photoreceptor types, distinguished by the expression of different rhodopsins. The rhodopsin Aaop3 (GPROP3) is expressed in the majority of the larval photoreceptors. There are two small clusters of photoreceptors located within the satellite and central stemmata that express the rhodopsin Aaop7 (GPROP7) instead of Aaop3. Electroretinogram analysis of transgenic Aaop7 Drosophila indicates that Aaop3 and Aaop7, both classified as long-wavelength rhodopsins, possess similar but not identical spectral properties. Light triggers an extensive translocation of Aaop3 from the photosensitive rhabdoms to the cytoplasmic compartment, whereas light-driven translocation of Aaop7 is limited. The results suggest that these photoreceptor cell types play distinct roles in larval vision. An additional component of the larval visual system is the adult compound eye, which starts to develop at the anterior face of the larval stemmata during the 1st instar stage. The photoreceptors of the developing compound eye show rhodopsin expression during the 4th larval instar stage, consistent with indications from previous reports that the adult compound eye contributes to larval and pupal visual capabilities. PMID:25750414

  7. Dynamic pattern of expression of dlin52, a member of the Myb/MuvB complex, during Drosophila development.

    PubMed

    Bhaskar, Pradeep Kumar; Mukherjee, Ashim; Mutsuddi, Mousumi

    2012-01-01

    The DREAM (DP, RB, E2F and MuvB) complex is required in humans to arrest the expression of cell cycle genes during quiescence. One of its members LIN52 has been isolated from the repressor complex but little is known about its molecular function. It has been reported recently that the serine residue 28 of LIN52 is phosphorylated by DYRK1A, and point mutation of this residue or down regulation of DYRK1A (which phosphorylates LIN52) leads to disruption of DREAM complex assembly, which is needed for G(0) arrest. Function of all the members of the dMyb complex (homologue of DREAM complex) in Drosophila melanogaster is not well characterized. We have studied the Drosophila orthologue of LIN52, known as dlin52, which is strongly conserved across various taxa from worms to human. dlin52 is reported to be present in a large protein complex containing important transcriptional regulators of cell proliferation and cell death like dE2F1, dMyb and dRbf. We have examined the expression of dlin52 transcripts and protein during development. Strong nuclear expression of dlin52 is seen in larval eye-antennal discs, brain, fat body, wing discs and salivary glands. dlin52 is abundantly expressed in endoreplicated tissues like salivary glands, fat body, and certain regions of the gut, and the nurse cells from adult ovaries. dlin52 is also expressed in the larval optic lobe, as well as in the developing neurons of ventral ganglion, indicating that this gene has an important role to play in cell cycle regulation and neuronal development. Robust expression of dlin52 protein was observed in quiescent cells like that of the imaginal cells of larval salivary gland, while marginal expression was seen in the germarium of adult ovary. Study of the spatial and temporal pattern of expression of this gene will help in better understanding of the function of this protein during various developmental processes. PMID:22178095

  8. Development of Johnston’s organ in Drosophila

    PubMed Central

    EBERL, DANIEL F.; BOEKHOFF-FALK, GRACE

    2012-01-01

    Hearing is a specialized mechanosensory modality that is refined during evolution to meet the particular requirements of different organisms. In the fruitfly, Drosophila, hearing is mediated by Johnston’s organ, a large chordotonal organ in the antenna that is exquisitely sensitive to the near-field acoustic signal of courtship songs generated by male wing vibration. We summarize recent progress in understanding the molecular genetic determinants of Johnston’s organ development and discuss surprising differences from other chordotonal organs that likely facilitate hearing. We outline novel discoveries of active processes that generate motion of the antenna for acute sensitivity to the stimulus. Finally, we discuss further research directions that would probe remaining questions in understanding Johnston’s organ development, function and evolution. PMID:17891726

  9. Regulatory mechanisms of EGFR signalling during Drosophila eye development.

    PubMed

    Malartre, Marianne

    2016-05-01

    EGFR signalling is a well-conserved signalling pathway playing major roles during development and cancers. This review explores what studying the EGFR pathway during Drosophila eye development has taught us in terms of the diversity of its regulatory mechanisms. This model system has allowed the identification of numerous positive and negative regulators acting at specific time and place, thus participating to the tight control of signalling. EGFR signalling regulation is achieved by a variety of mechanisms, including the control of ligand processing, the availability of the receptor itself and the transduction of the cascade in the cytoplasm. Ultimately, the transcriptional responses contribute to the establishment of positive and negative feedback loops. The combination of these multiple mechanisms employed to regulate the EGFR pathway leads to specific cellular outcomes involved in functions as diverse as the acquisition of cell fate, proliferation, survival, adherens junction remodelling and morphogenesis. PMID:26935860

  10. Morphological diversity and development of glia in Drosophila.

    PubMed

    Hartenstein, Volker

    2011-09-01

    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. On the basis of 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

  11. Suitability of monotypic and mixed diets for Anopheles hermsi larval development.

    PubMed

    Beasley, Donald A; Walton, William E

    2016-06-01

    The developmental time and survival to eclosion of Anopheles hermsi Barr & Guptavanij fed monotypic and mixed diets of ten food types were examined in laboratory studies. Larvae fed monotypic diets containing animal detritus (freeze-dried rotifers, freeze-dried Daphnia pulicaria, and TetraMin® fish food flakes) and the mixotrophic protistan Cryptomonas ovata developed faster and survived better than larvae that were fed other monotypic diets. Survival to adulthood of larvae fed several concentrations of the diatom Planothidium (=Achnanthes) lanceolatum was poor (<13%) and larval development time was approximately twice that of larvae fed TetraMin® fish food flakes, the standard laboratory diet. Larvae fed monotypic diets containing prokaryotes (bacteria [Bacillus cereus] and cyanobacteria [Oscillatoria prolifera]) and brewer's yeast (Saccharomyces cerevisiae) failed to survive beyond the 1(st) and 2(nd) instar, respectively. Larvae fed only chlorophytes, single-celled Chlamydomonas reinhardtii and filamentous Spirogyra communis, failed to complete larval development, regardless of the concentration tested. Cohorts fed a combination of food types (mixed diets) usually developed better than cohorts fed monotypic diets. Food types that failed to support complete development when fed alone often facilitated development to adulthood when fed in combination with food types containing >1% C20 polyunsaturated fatty acids as total fat, but regardless of essential fatty acid content, algae that produced mucilage and filaments that sank out of the feeding zone were poor quality diets. PMID:27232128

  12. Tissue landscape alters adjacent cell fates during Drosophila egg development

    PubMed Central

    Manning, Lathiena; Weideman, Ann Marie; Peercy, Bradford; Starz-Gaiano, Michelle

    2015-01-01

    Extracellular signaling molecules control many biological processes, but the influence of tissue architecture on the local concentrations of these factors is unclear. Here we examine this issue in the Drosophila egg chamber, where two anterior cells secrete Unpaired (Upd) to activate Signal Transducer and Activator of Transcription (STAT) signaling in the epithelium. High STAT signaling promotes cell motility. Genetic analysis shows that all cells near the Upd source can respond. However, using upright imaging, we show surprising asymmetries in STAT activation patterns, suggesting that some cells experience different Upd levels than predicted by their location. We develop a three-dimensional mathematical model to characterize the spatio-temporal distribution of the activator. Simulations show that irregular tissue domains can produce asymmetric distributions of Upd, consistent with results in vivo. Mutant analysis substantiates this idea. We conclude that cellular landscape can heavily influence the effect of diffusible activators and should be more widely considered. PMID:26082073

  13. Quantifying and predicting Drosophila larvae crawling phenotypes.

    PubMed

    Günther, Maximilian N; Nettesheim, Guilherme; Shubeita, George T

    2016-01-01

    The fruit fly Drosophila melanogaster is a widely used model for cell biology, development, disease, and neuroscience. The fly's power as a genetic model for disease and neuroscience can be augmented by a quantitative description of its behavior. Here we show that we can accurately account for the complex and unique crawling patterns exhibited by individual Drosophila larvae using a small set of four parameters obtained from the trajectories of a few crawling larvae. The values of these parameters change for larvae from different genetic mutants, as we demonstrate for fly models of Alzheimer's disease and the Fragile X syndrome, allowing applications such as genetic or drug screens. Using the quantitative model of larval crawling developed here we use the mutant-specific parameters to robustly simulate larval crawling, which allows estimating the feasibility of laborious experimental assays and aids in their design. PMID:27323901

  14. Quantifying and predicting Drosophila larvae crawling phenotypes

    PubMed Central

    Günther, Maximilian N.; Nettesheim, Guilherme; Shubeita, George T.

    2016-01-01

    The fruit fly Drosophila melanogaster is a widely used model for cell biology, development, disease, and neuroscience. The fly’s power as a genetic model for disease and neuroscience can be augmented by a quantitative description of its behavior. Here we show that we can accurately account for the complex and unique crawling patterns exhibited by individual Drosophila larvae using a small set of four parameters obtained from the trajectories of a few crawling larvae. The values of these parameters change for larvae from different genetic mutants, as we demonstrate for fly models of Alzheimer’s disease and the Fragile X syndrome, allowing applications such as genetic or drug screens. Using the quantitative model of larval crawling developed here we use the mutant-specific parameters to robustly simulate larval crawling, which allows estimating the feasibility of laborious experimental assays and aids in their design. PMID:27323901

  15. Larval spicules, cilia, and symmetry as remnants of indirect development in the direct developing sea urchin Heliocidaris erythrogramma.

    PubMed

    Emlet, R B

    1995-02-01

    Nonfeeding larvae of the echinoid Heliocidaris erythrogramma were raised in culture and examined for expression of a larval skeleton and for the arrangement of the ciliated band. Opaque larvae were fixed, cleared, and examined under polarized light for evidence of calcification. By 35 hr after fertilization (at 22 degrees C), a pair of triradiate spicules was present at the posterior end of the larvae. Each member of this pair formed a fenestrated spicule as it grew laterally. This pair and another pair which formed subsequently, were arranged across a plane of bilateral symmetry orthagonal to the juvenile oral aboral axis. These paired larval spicules can be identified as reduced expressions of postoral and posterodorsal rods found in plutei, and their expression indicates that the juvenile rudiment of H. erythrogramma forms on the left side and that larval body axes are conserved in this modified larva. By 44 hr the ciliated band formed as an incomplete transverse loop of three segments at the posterior end and on the dorsal surface of the ovoid larva. Cilia in these segments grew to lengths of 45-50 microns, longer than other swimming and feeding cilia reported for echinoderm larvae. Band segments are interpreted as expressions of epaulettes (specialized swimming bands) rather than the feeding ciliated band of the pluteus. The ciliated band segments and the larval spicules are both bilaterally symmetrical with respect to the same plane and indicate conserved larval bilateral symmetry despite the major asymmetry of the fates of cells on either side of this plane in their contribution to juvenile development. PMID:7875367

  16. Drosophila ptip is essential for anterior/posterior patterning in development and interacts with the PcG and trxG pathways

    PubMed Central

    Fang, Ming; Ren, Hongyan; Liu, Jiabin; Cadigan, Ken M.; Patel, Sanjeevkumar R.; Dressler, Gregory R.

    2009-01-01

    Summary Development of the fruit fly Drosophila depends in part on epigenetic regulation carried out by the concerted actions of the Polycomb and Trithorax group of proteins, many of which are associated with histone methyltransferase activity. Mouse PTIP is part of a histone H3K4 methyltransferase complex and contains six BRCT domains and a glutamine-rich region. In this article, we describe an essential role for the Drosophila ortholog of the mammalian Ptip (Paxip1) gene in early development and imaginal disc patterning. Both maternal and zygotic ptip are required for segmentation and axis patterning during larval development. Loss of ptip results in a decrease in global levels of H3K4 methylation and an increase in the levels of H3K27 methylation. In cell culture, Drosophila ptip is required to activate homeotic gene expression in response to the derepression of Polycomb group genes. Activation of developmental genes is coincident with PTIP protein binding to promoter sequences and increased H3K4 trimethylation. These data suggest a highly conserved function for ptip in epigenetic control of development and differentiation. PMID:19429789

  17. Urbanization Increases Aedes albopictus Larval Habitats and Accelerates Mosquito Development and Survivorship

    PubMed Central

    Li, Yiji; Kamara, Fatmata; Zhou, Guofa; Puthiyakunnon, Santhosh; Li, Chunyuan; Liu, Yanxia; Zhou, Yanhe; Yao, Lijie; Yan, Guiyun; Chen, Xiao-Guang

    2014-01-01

    Introduction Aedes albopictus is a very invasive and aggressive insect vector that causes outbreaks of dengue fever, chikungunya disease, and yellow fever in many countries. Vector ecology and disease epidemiology are strongly affected by environmental changes. Urbanization is a worldwide trend and is one of the most ecologically modifying phenomena. The purpose of this study is to determine how environmental changes due to urbanization affect the ecology of Aedes albopictus. Methods Aquatic habitats and Aedes albopictus larval population surveys were conducted from May to November 2013 in three areas representing rural, suburban, and urban settings in Guangzhou, China. Ae. albopictus adults were collected monthly using BG-Sentinel traps. Ae. albopictus larva and adult life-table experiments were conducted with 20 replicates in each of the three study areas. Results The urban area had the highest and the rural area had the lowest number of aquatic habitats that tested positive for Ae. albopictus larvae. Densities in the larval stages varied among the areas, but the urban area had almost two-fold higher densities in pupae and three-fold higher in adult populations compared with the suburban and rural areas. Larvae developed faster and the adult emergence rate was higher in the urban area than in suburban and rural areas. The survival time of adult mosquitoes was also longer in the urban area than it was in suburban and rural areas. Study regions, surface area, water depth, water clearance, surface type, and canopy coverage were important factors associated with the presence of Ae. albopictus larvae. Conclusions Urbanization substantially increased the density, larval development rate, and adult survival time of Ae. albopictus, which in turn potentially increased the vector capacity, and therefore, disease transmissibility. Mosquito ecology and its correlation with dengue virus transmission should be compared in different environmental settings. PMID:25393814

  18. Effect of Two Oil Dispersants on Larval Grass Shrimp (Palaemonetes pugio) Development.

    NASA Astrophysics Data System (ADS)

    Betancourt, P.; Key, P. B.; Chung, K. W.; DeLorenzo, M. E.

    2015-12-01

    The study focused on the effects that two oil dispersants, Corexit® EC9500A and Finasol® OSR52, have on the development of larval grass shrimp, (Palaemonetes pugio). The hypothesis was that Finasol would have a greater effect on larval grass shrimp development than Corexit. The experiment was conducted using 300 grass shrimp larvae that were 24 hours old. Each larva was exposed individually. In total, five sub-lethal concentrations were tested for each dispersant (control, 1.25, 2.50, 5.0,10.0 mg/L). The larvae were exposed for five days then transferred to clean seawater until metamorphosis into the juvenile stage. Key data measurements recorded included number of days to become juveniles, number of instars, length, dry weight, and mortality. Data from exposed shrimp was compared to the results of the control for each dispersant concentration. Corexit and Finasol exposure treatments of 5 mg/L and 10 mg/L showed significantly higher values for number of days and number of instars to reach juvenile status than values obtained from unexposed, control shrimp. Overall, mortality was higher in the Finasol treatments but the two dispersants did not respond significantly different from one another. Future studies are needed to determine the long term effects of dispersant exposure on all grass shrimp life stages and how any dispersant exposure impacts grass shrimp populations. Grass shrimp serve as excellent toxicity indicators of estuaries, and further studies will help to develop better oil spill mitigation techniques.

  19. Growth and development of larval green frogs (Rana clamitans) exposed to multiple doses of an insecticide

    USGS Publications Warehouse

    Boone, M.D.; Bridges, C.M.; Rothermel, B.B.

    2001-01-01

    Our objective was to determine how green frogs (Rana clamitans) are affected by multiple exposures to a sublethal level of the carbamate insecticide, carbaryl, in outdoor ponds. Tadpoles were added to 1,000-1 ponds at a low or high density which were exposed to carbaryl 0, 1, 2, or 3 times. Length of the larval period, mass, developmental stage, tadpole survival, and proportion metamorphosed were used to determine treatment effects. The frequency of dosing affected the proportion of green frogs that reached metamorphosis and the developmental stage of tadpoles. Generally, exposure to carbaryl increased rates of metamorphosis and development. The effect of the frequency of carbaryl exposure on development varied with the density treatment; the majority of metamorphs and the most developed tadpoles came from high-density ponds exposed to carbaryl 3 times. This interaction suggests that exposure to carbaryl later in the larval period stimulated metamorphosis, directly or indirectly, under high-density conditions. Our study indicates that exposure to a contaminant can lead to early initiation of metamorphosis and that natural biotic factors can mediate the effects of a contaminant in the environment.

  20. Aspects of Embryonic and Larval Development in Bighead Carp Hypophthalmichthys nobilis and Silver Carp Hypophthalmichthys molitrix

    PubMed Central

    George, Amy E.; Chapman, Duane C.

    2013-01-01

    As bighead carp Hypophthalmichthysnobilis and silver carp H. molitrix (the bigheaded carps) are poised to enter the Laurentian Great Lakes and potentially damage the region’s economically important fishery, information on developmental rates and behaviors of carps is critical to assessing their ability to establish sustainable populations within the Great Lakes basin. In laboratory experiments, the embryonic and larval developmental rates, size, and behaviors of bigheaded carp were tracked at two temperature treatments, one “cold” and one “warm”. Developmental rates were computed using previously described stages of development and the cumulative thermal unit method. Both species have similar thermal requirements, with a minimum developmental temperature for embryonic stages of 12.1° C for silver carp and 12.9° C for bighead carp, and 13.3° C for silver carp larval stages and 13.4° C for bighead carp larval stages. Egg size differed among species and temperature treatments, as egg size was larger in bighead carp, and “warm" temperature treatments. The larvae started robust upwards vertical swimming immediately after hatching, interspersed with intervals of sinking. Vertical swimming tubes were used to measure water column distribution, and ascent and descent rates of vertically swimming fish. Water column distribution and ascent and descent rates changed with ontogeny. Water column distribution also showed some diel periodicity. Developmental rates, size, and behaviors contribute to the drift distance needed to fulfill the early life history requirements of bigheaded carps and can be used in conjunction with transport information to assess invasibility of a river. PMID:23967350

  1. Aspects of embryonic and larval development in bighead carp Hypophthalmichthys nobilis and silver carp Hypophthalmichthys molitrix

    USGS Publications Warehouse

    George, Amy E.; Chapman, Duane C.

    2013-01-01

    As bighead carp Hypophthalmichthys nobilis and silver carp H. molitrix (the bigheaded carps) are poised to enter the Laurentian Great Lakes and potentially damage the region’s economically important fishery, information on developmental rates and behaviors of carps is critical to assessing their ability to establish sustainable populations within the Great Lakes basin. In laboratory experiments, the embryonic and larval developmental rates, size, and behaviors of bigheaded carp were tracked at two temperature treatments, one “cold” and one “warm”. Developmental rates were computed using previously described stages of development and the cumulative thermal unit method. Both species have similar thermal requirements, with a minimum developmental temperature for embryonic stages of 12.1° C for silver carp and 12.9° C for bighead carp, and 13.3° C for silver carp larval stages and 13.4° C for bighead carp larval stages. Egg size differed among species and temperature treatments, as egg size was larger in bighead carp, and “warm" temperature treatments. The larvae started robust upwards vertical swimming immediately after hatching, interspersed with intervals of sinking. Vertical swimming tubes were used to measure water column distribution, and ascent and descent rates of vertically swimming fish. Water column distribution and ascent and descent rates changed with ontogeny. Water column distribution also showed some diel periodicity. Developmental rates, size, and behaviors contribute to the drift distance needed to fulfill the early life history requirements of bigheaded carps and can be used in conjunction with transport information to assess invasibility of a river.

  2. Expression of Drosophila Forkhead Transcription Factors During Kidney Development

    PubMed Central

    Baek, Jeong-In; Choi, Soo Young; Chacon-Heszele, Maria F.; Zuo, Xiaofeng; Lipschutz, Joshua H.

    2014-01-01

    The Drosophila forkhead (Dfkh) family of transcription factors has over 40 family members. One Dfkh family member, BF2 (aka FoxD1), has been shown, by targeted disruption, to be essential for kidney development. In order to determine if other Dfkh family members were involved in kidney development and to search for new members of this family, reverse transcriptase polymerase chain reaction (RT-PCR) was performed using degenerate primers of the consensus sequence of the DNA binding domain of this family and developing rat kidney RNA. The RT-PCR product was used to probe RNA from a developing rat kidney (neonatal), from a 20-day old kidney, and from an adult kidney. The RT-PCR product hybridized only to a developing kidney RNA transcript of ~2.3 kb (the size of BF2). A lambda gt10 mouse neonatal kidney library was then screened, using the above-described RT-PCR product as a probe. Three lambda phage clones were isolated that strongly hybridized to the RT-PCR probe. Sequencing of the RT-PCR product and the lambda phage clones isolated from the developing kidney library revealed Dfkh BF2. In summary, only Dfkh family member BF2, which has already been shown to be essential for nephrogenesis, was identified in our screen and no other candidate Dfkh family members were identified. PMID:24491558

  3. Effect of condensed tannins on egg hatching and larval development of Trichostrongylus colubriformis in vitro.

    PubMed

    Molan, A L; Waghorn, G C; McNabb, W C

    2002-01-19

    The effects of condensed tannins extracted from seven forages on the viability of the eggs and first stage (L1) larvae of the sheep nematode Trichostrongylus colubriformis were evaluated in in vitro assays. The extracts of condensed tannins were obtained from Lotus pedunculatus (LP), Lotus corniculatus (LC), sulla (Hedysarum coronarium), sainfoin (Onobrychus viciifolia), Dorycnium pentaphylum (DP), Dorycnium rectum (DR) and dock (Rumex obtusifolius). Extracts containing 200 to 500 microg/ml reduced the proportion of eggs that hatched. The larval development assay was used to evaluate the effect of the extracts on the development of either eggs or L1 larvae to L3 infective larvae. Development was allowed to proceed for seven days by which time the larvae in control incubations had reached the infective L3 stage. Extracts containing 200 microg/ml from LP, DP, DR or dock prevented egg development, and only 11, 8 and 2 per cent of the eggs developed to L3 larvae with extracts from LC, sulla and sainfoin, respectively. When the concentration was 400 microg/ml no eggs developed to L3 larvae. The addition of the extracts after hatching also inhibited the development of L1 to L3 larvae; 200 microg/ml extracted from LP, LC, sulla, sainfoin, DP, DR and dock resulted in only 14, 18, 17, 15, 14, 16 and 4 per cent of L1 larvae developing to the L3 stage compared with 85 per cent for controls, and 400 microg/ml further reduced the development of L1 larvae. Statistical analyses showed that when the extracts were added before hatching they were significantly (P<0.001) more effective at inhibiting the larval development than when they were added after hatching. The condensed tannins from dock had the greatest inhibitory effect on egg development followed by the tannins from DR, sainfoin, DP, LP, sulla and LC. PMID:11837588

  4. Embryonic and larval development of the host sea anemones Entacmaea quadricolor and Heteractis crispa.

    PubMed

    Scott, Anna; Harrison, Peter L

    2007-10-01

    Little information is available on the sexual reproductive biology of anemones that provide essential habitat for anemonefish. Here we provide the first information on the surface ultrastructural and morphological changes during development of the embryos and planula larvae of Entacmaea quadricolor and Heteractis crispa, using light and scanning electron microscopy. Newly spawned eggs of E. quadricolor and H. crispa averaged 794 microm and 589 microm diameter, respectively, and were covered by many spires of microvilli that were evenly distributed over the egg surface, except for a single bare patch. Eggs of both species contained abundant zooxanthellae when spawned, indicating vertical transmission of symbionts. Fertilization was external, and the resulting embryos displayed superficial cleavage. As development continued, individual blastomeres became readily distinguishable and a round-to-ovoid blastula was formed, which flattened with further divisions. The edges of the blastula thickened, creating a concave-convex dish-shaped gastrula. The outer margins of the gastrula appeared to roll inward, leading to the formation of an oral pore and a ciliated planula larva. Larval motility and directional movement were first observed 36 h after spawning. E. quadricolor larval survival remained high during the first 4 d after spawning, then decreased rapidly. PMID:17928518

  5. Rearing Tenebrio molitor in BLSS: Dietary fiber affects larval growth, development, and respiration characteristics

    NASA Astrophysics Data System (ADS)

    Li, Leyuan; Stasiak, Michael; Li, Liang; Xie, Beizhen; Fu, Yuming; Gidzinski, Danuta; Dixon, Mike; Liu, Hong

    2016-01-01

    Rearing of yellow mealworm (Tenebrio molitor L.) will provide good animal nutrition for astronauts in a bioregenerative life support system. In this study, growth and biomass conversion data of T. molitor larvae were tested for calculating the stoichiometric equation of its growth. Result of a respiratory quotient test proved the validity of the equation. Fiber had the most reduction in mass during T. molitor‧s consumption, and thus it is speculated that fiber is an important factor affecting larval growth of T. molitor. In order to further confirm this hypothesis and find out a proper feed fiber content, T. molitor larvae were fed on diets with 4 levels of fiber. Larval growth, development and respiration in each group were compared and analyzed. Results showed that crude-fiber content of 5% had a significant promoting effect on larvae in early instars, and is beneficial for pupa eclosion. When fed on feed of 5-10% crude-fiber, larvae in later instars reached optimal levels in growth, development and respiration. Therefore, we suggest that crude fiber content in feed can be controlled within 5-10%, and with the consideration of food palatability, a crude fiber of 5% is advisable.

  6. Transcriptome Profiling Identifies Multiplexin as a Target of SAGA Deubiquitinase Activity in Glia Required for Precise Axon Guidance During Drosophila Visual Development

    PubMed Central

    Ma, Jingqun; Brennan, Kaelan J.; D’Aloia, Mitch R.; Pascuzzi, Pete E.; Weake, Vikki M.

    2016-01-01

    The Spt-Ada-Gcn5 Acetyltransferase (SAGA) complex is a transcriptional coactivator with histone acetylase and deubiquitinase activities that plays an important role in visual development and function. In Drosophila melanogaster, four SAGA subunits are required for the deubiquitination of monoubiquitinated histone H2B (ubH2B): Nonstop, Sgf11, E(y)2, and Ataxin 7. Mutations that disrupt SAGA deubiquitinase activity cause defects in neuronal connectivity in the developing Drosophila visual system. In addition, mutations in SAGA result in the human progressive visual disorder spinocerebellar ataxia type 7 (SCA7). Glial cells play a crucial role in both the neuronal connectivity defect in nonstop and sgf11 flies, and in the retinal degeneration observed in SCA7 patients. Thus, we sought to identify the gene targets of SAGA deubiquitinase activity in glia in the Drosophila larval central nervous system. To do this, we enriched glia from wild-type, nonstop, and sgf11 larval optic lobes using affinity-purification of KASH-GFP tagged nuclei, and then examined each transcriptome using RNA-seq. Our analysis showed that SAGA deubiquitinase activity is required for proper expression of 16% of actively transcribed genes in glia, especially genes involved in proteasome function, protein folding and axon guidance. We further show that the SAGA deubiquitinase-activated gene Multiplexin (Mp) is required in glia for proper photoreceptor axon targeting. Mutations in the human ortholog of Mp, COL18A1, have been identified in a family with a SCA7-like progressive visual disorder, suggesting that defects in the expression of this gene in SCA7 patients could play a role in the retinal degeneration that is unique to this ataxia. PMID:27261002

  7. Transcriptome Profiling Identifies Multiplexin as a Target of SAGA Deubiquitinase Activity in Glia Required for Precise Axon Guidance During Drosophila Visual Development.

    PubMed

    Ma, Jingqun; Brennan, Kaelan J; D'Aloia, Mitch R; Pascuzzi, Pete E; Weake, Vikki M

    2016-01-01

    The Spt-Ada-Gcn5 Acetyltransferase (SAGA) complex is a transcriptional coactivator with histone acetylase and deubiquitinase activities that plays an important role in visual development and function. In Drosophila melanogaster, four SAGA subunits are required for the deubiquitination of monoubiquitinated histone H2B (ubH2B): Nonstop, Sgf11, E(y)2, and Ataxin 7. Mutations that disrupt SAGA deubiquitinase activity cause defects in neuronal connectivity in the developing Drosophila visual system. In addition, mutations in SAGA result in the human progressive visual disorder spinocerebellar ataxia type 7 (SCA7). Glial cells play a crucial role in both the neuronal connectivity defect in nonstop and sgf11 flies, and in the retinal degeneration observed in SCA7 patients. Thus, we sought to identify the gene targets of SAGA deubiquitinase activity in glia in the Drosophila larval central nervous system. To do this, we enriched glia from wild-type, nonstop, and sgf11 larval optic lobes using affinity-purification of KASH-GFP tagged nuclei, and then examined each transcriptome using RNA-seq. Our analysis showed that SAGA deubiquitinase activity is required for proper expression of 16% of actively transcribed genes in glia, especially genes involved in proteasome function, protein folding and axon guidance. We further show that the SAGA deubiquitinase-activated gene Multiplexin (Mp) is required in glia for proper photoreceptor axon targeting. Mutations in the human ortholog of Mp, COL18A1, have been identified in a family with a SCA7-like progressive visual disorder, suggesting that defects in the expression of this gene in SCA7 patients could play a role in the retinal degeneration that is unique to this ataxia. PMID:27261002

  8. The Friend of GATA proteins U-shaped, FOG-1, and FOG-2 function as negative regulators of blood, heart, and eye development in Drosophila

    PubMed Central

    Fossett, Nancy; Tevosian, Sergei G.; Gajewski, Kathleen; Zhang, Qian; Orkin, Stuart H.; Schulz, Robert A.

    2001-01-01

    Friend of GATA (FOG) proteins regulate GATA factor-activated gene transcription. During vertebrate hematopoiesis, FOG and GATA proteins cooperate to promote erythrocyte and megakaryocyte differentiation. The Drosophila FOG homologue U-shaped (Ush) is expressed similarly in the blood cell anlage during embryogenesis. During hematopoiesis, the acute myeloid leukemia 1 homologue Lozenge and Glial cells missing are required for the production of crystal cells and plasmatocytes, respectively. However, additional factors have been predicted to control crystal cell proliferation. In this report, we show that Ush is expressed in hemocyte precursors and plasmatocytes throughout embryogenesis and larval development, and the GATA factor Serpent is essential for Ush embryonic expression. Furthermore, loss of ush function results in an overproduction of crystal cells, whereas forced expression of Ush reduces this cell population. Murine FOG-1 and FOG-2 also can repress crystal cell production, but a mutant version of FOG-2 lacking a conserved motif that binds the corepressor C-terminal binding protein fails to affect the cell lineage. The GATA factor Pannier (Pnr) is required for eye and heart development in Drosophila. When Ush, FOG-1, FOG-2, or mutant FOG-2 is coexpressed with Pnr during these developmental processes, severe eye and heart phenotypes result, consistent with a conserved negative regulation of Pnr function. These results indicate that the fly and mouse FOG proteins function similarly in three distinct cellular contexts in Drosophila, but may use different mechanisms to regulate genetic events in blood vs. cardial or eye cell lineages. PMID:11404479

  9. Spatial expression of transcription factors in Drosophila embryonic organ development

    PubMed Central

    2013-01-01

    Background Site-specific transcription factors (TFs) bind DNA regulatory elements to control expression of target genes, forming the core of gene regulatory networks. Despite decades of research, most studies focus on only a small number of TFs and the roles of many remain unknown. Results We present a systematic characterization of spatiotemporal gene expression patterns for all known or predicted Drosophila TFs throughout embryogenesis, the first such comprehensive study for any metazoan animal. We generated RNA expression patterns for all 708 TFs by in situ hybridization, annotated the patterns using an anatomical controlled vocabulary, and analyzed TF expression in the context of organ system development. Nearly all TFs are expressed during embryogenesis and more than half are specifically expressed in the central nervous system. Compared to other genes, TFs are enriched early in the development of most organ systems, and throughout the development of the nervous system. Of the 535 TFs with spatially restricted expression, 79% are dynamically expressed in multiple organ systems while 21% show single-organ specificity. Of those expressed in multiple organ systems, 77 TFs are restricted to a single organ system either early or late in development. Expression patterns for 354 TFs are characterized for the first time in this study. Conclusions We produced a reference TF dataset for the investigation of gene regulatory networks in embryogenesis, and gained insight into the expression dynamics of the full complement of TFs controlling the development of each organ system. PMID:24359758

  10. Evolved differences in larval social behavior mediated by novel pheromones

    PubMed Central

    Mast, Joshua D; De Moraes, Consuelo M; Alborn, Hans T; Lavis, Luke D; Stern, David L

    2014-01-01

    Pheromones, chemical signals that convey social information, mediate many insect social behaviors, including navigation and aggregation. Several studies have suggested that behavior during the immature larval stages of Drosophila development is influenced by pheromones, but none of these compounds or the pheromone-receptor neurons that sense them have been identified. Here we report a larval pheromone-signaling pathway. We found that larvae produce two novel long-chain fatty acids that are attractive to other larvae. We identified a single larval chemosensory neuron that detects these molecules. Two members of the pickpocket family of DEG/ENaC channel subunits (ppk23 and ppk29) are required to respond to these pheromones. This pheromone system is evolving quickly, since the larval exudates of D. simulans, the sister species of D. melanogaster, are not attractive to other larvae. Our results define a new pheromone signaling system in Drosophila that shares characteristics with pheromone systems in a wide diversity of insects. DOI: http://dx.doi.org/10.7554/eLife.04205.001 PMID:25497433

  11. Ire1 supports normal ER differentiation in developing Drosophila photoreceptors

    PubMed Central

    Xu, Zuyuan; Chikka, Madhusudana Rao; Xia, Hongai; Ready, Donald F.

    2016-01-01

    ABSTRACT The endoplasmic reticulum (ER) serves virtually all aspects of cell physiology and, by pathways that are incompletely understood, is dynamically remodeled to meet changing cell needs. Inositol-requiring enzyme 1 (Ire1), a conserved core protein of the unfolded protein response (UPR), participates in ER remodeling and is particularly required during the differentiation of cells devoted to intense secretory activity, so-called ‘professional’ secretory cells. Here, we characterize the role of Ire1 in ER differentiation in the developing Drosophila compound eye photoreceptors (R cells). As part of normal development, R cells take a turn as professional secretory cells with a massive secretory effort that builds the photosensitive membrane organelle, the rhabdomere. We find rough ER sheets proliferate as rhabdomere biogenesis culminates, and Ire1 is required for normal ER differentiation. Ire1 is active early in R cell development and is required in anticipation of peak biosynthesis. Without Ire1, the amount of rough ER sheets is strongly reduced and the extensive cortical ER network at the rhabdomere base, the subrhabdomere cisterna (SRC), fails. Instead, ER proliferates in persistent and ribosome-poor tubular tangles. A phase of Ire1 activity early in R cell development thus shapes dynamic ER. PMID:26787744

  12. Cloning of aquaporin-1 of the blue crab, Callinectes sapidus: its expression during the larval development in hyposalinity

    PubMed Central

    2012-01-01

    Background Ontogenetic variation in salinity adaptation has been noted for the blue crab, Callinectes sapidus, which uses the export strategy for larval development: females migrate from the estuaries to the coast to spawn, larvae develop in the ocean, and postlarvae (megalopae) colonize estuarine areas. We hypothesized that C. sapidus larvae may be stenohaline and have limited osmoregulatory capacity which compromises their ability to survive in lower salinity waters. We tested this hypothesis using hatchery-raised larvae that were traceable to specific life stages. In addition, we aimed to understand the possible involvement of AQP-1 in salinity adaptation during larval development and during exposure to hyposalinity. Results A full-length cDNA sequence of aquaporin (GenBank JQ970426) was isolated from the hypodermis of the blue crab, C. sapidus, using PCR with degenerate primers and 5′ and 3′ RACE. The open reading frame of CasAQP-1 consists of 238 amino acids containing six helical structures and two NPA motifs for the water pore. The expression pattern of CasAQP-1 was ubiquitous in cDNAs from all tissues examined, although higher in the hepatopancreas, thoracic ganglia, abdominal muscle, and hypodermis and lower in the antennal gland, heart, hemocytes, ovary, eyestalk, brain, hindgut, Y-organs, and gill. Callinectes larvae differed in their capacity to molt in hyposalinity, as those at earlier stages from Zoea (Z) 1 to Z4 had lower molting rates than those from Z5 onwards, as compared to controls kept in 30 ppt water. No difference was found in the survival of larvae held at 15 and 30 ppt. CasAQP-1 expression differed with ontogeny during larval development, with significantly higher expression at Z1-2, compared to other larval stages. The exposure to 15 ppt affected larval-stage dependent CasAQP-1 expression which was significantly higher in Z2- 6 stages than the other larval stages. Conclusions We report the ontogenetic variation in CasAQP-1

  13. Toxicity of endosulfan on embryo-larval development of the South American toad Rhinella arenarum.

    PubMed

    Svartz, Gabriela V; Wolkowicz, Ianina R Hutler; Coll, Cristina S Pérez

    2014-04-01

    Endosulfan is a widely used pesticide despite its extreme toxicity to a variety of taxa and its worldwide ban. The aim of the present study was to evaluate the acute and chronic toxicity of endosulfan on the embryonic-larval development of the common South American toad Rhinella arenarum. The results showed that lethal and sublethal effects increased with concentration and exposure time. The sensitivity to endosulfan increased during the larval period, the complete operculum stage (S.25) being the most sensitive (504-h median lethal concentration [LC50] = 0.01 mg endosulfan/L; 10% lethal concentration [LC10] = 0.004 mg endosulfan/L). Endosulfan exposure caused morphological abnormalities such as general underdevelopment, edema, gill malformations, and cellular dissociation as well as neurotoxicity. Our results also showed that larvae exposed to concentrations of 0.005 mg endosulfan/L and 0.01 mg endosulfan/L completed metamorphosis earlier than controls, but with underdevelopment. The 240-h teratogenic index was 6.13, implying a high risk for embryos to be malformed in the absence of significant embryonic lethality. Because the hazard quotients for chronic exposure were over 1, the level of concern value and toxicity endpoints obtained in the present study for R. arenarum occurred at concentrations lower than the levels of endosulfan reported in the environment, this pesticide should be considered a potential risk for this species. PMID:24375551

  14. Particle motion is broadly represented in the vestibular medulla of the bullfrog across larval development.

    PubMed

    Simmons, Andrea Megela; Flores, Victoria

    2012-04-01

    In their shallow-water habitats, bullfrog (Rana catesbeiana) tadpoles are exposed to both underwater and airborne sources of acoustic stimulation. We probed the representation of underwater particle motion throughout the tadpole's dorsal medulla to determine its spatial extent over larval life. Using neurobiotin-filled micropipettes, we recorded neural activity to z-axis particle motion (frequencies of 40-200 Hz) in the medial vestibular nucleus, lateral vestibular nucleus, dorsal medullary nucleus (DMN), and along the dorsal arcuate pathway. Sensitivity was comparable in the medial and lateral vestibular nuclei, with estimated thresholds between 0.016 and 12.5 μm displacement. Neither best responding frequency nor estimated threshold varied significantly over larval stage. Transport of neurobiotin from active recording sites was also stable over development. The DMN responded poorly to z-axis particle motion, but did respond to low-frequency pressure stimulation. These data suggest that particle motion is represented widely and stably in the tadpole's vestibular medulla. This is in marked contrast to the representation of pressure stimulation in the auditory midbrain, where a transient "deaf period" of non-responsiveness and decreased connectivity occurs immediately prior to metamorphic climax. We suggest that, in bullfrogs, sensitivity to particle motion and to pressure follows different developmental trajectories. PMID:22198742

  15. lin-35/Rb cooperates with the SWI/SNF complex to control Caenorhabditis elegans larval development.

    PubMed Central

    Cui, Mingxue; Fay, David S; Han, Min

    2004-01-01

    Null mutations in lin-35, the Caenorhabditis elegans ortholog of the mammalian Rb protein, cause no obvious morphological defects. Using a genetic approach to identify genes that may function redundantly with lin-35, we have isolated a mutation in the C. elegans psa-1 gene. lin-35; psa-1 double mutants display severe developmental defects leading to early larval arrest and adult sterility. The psa-1 gene has previously been shown to encode a C. elegans homolog of yeast SWI3, a critical component of the SWI/SNF complex, and has been shown to regulate asymmetric cell divisions during C. elegans development. We observed strong genetic interactions between psa-1 and lin-35 as well as a subset of the class B synMuv genes that include lin-37 and lin-9. Loss-of-function mutations in lin-35, lin-37, and lin-9 strongly enhanced the defects of asymmetric T cell division associated with a psa-1 mutation. Our results suggest that LIN-35/Rb and a certain class B synMuv proteins collaborate with the SWI/SNF protein complex to regulate the T cell division as well as other events essential for larval growth. PMID:15280233

  16. Effects of temperature on embryonic and early larval growth and development in the rough-skinned newt (Taricha granulosa).

    PubMed

    Smith, Geoffrey D; Hopkins, Gareth R; Mohammadi, Shabnam; M Skinner, Heather; Hansen, Tyler; Brodie, Edmund D; French, Susannah S

    2015-07-01

    We investigated the effects of temperature on the growth and development of embryonic and early larval stages of a western North American amphibian, the rough-skinned newt (Taricha granulosa). We assigned newt eggs to different temperatures (7, 14, or 21°C); after hatching, we re-assigned the newt larvae into the three different temperatures. Over the course of three to four weeks, we measured total length and developmental stage of the larvae. Our results indicated a strong positive relationship over time between temperature and both length and developmental stage. Importantly, individuals assigned to cooler embryonic temperatures did not achieve the larval sizes of individuals from the warmer embryonic treatments, regardless of larval temperature. Our investigation of growth and development at different temperatures demonstrates carry-over effects and provides a more comprehensive understanding of how organisms respond to temperature changes during early development. PMID:25965021

  17. Small Peptides as Newcomers in the Control of Drosophila Development.

    PubMed

    Zanet, J; Chanut-Delalande, H; Plaza, Serge; Payre, Francios

    2016-01-01

    Throughout the last century, studies using the fruit fly have contributed to the discovery of many key genetic elements that control animal development. Recent work has shed light on an unexpectedly large number of RNAs that lack the classical hallmarks of protein-coding genes and are thus referred to as noncoding RNAs. However, there is mounting evidence that both mRNA and noncoding RNAs often contain small open reading frames (sORFs/smORFs), which can be translated into peptides. While genome-wide profiling supports a pervasive translation of these noncanonical sORF/smORF/SEP peptides, their functions remain poorly understood. Here, we review recent data obtained in Drosophila demonstrating the overlooked role of smORF peptides in the control of development and adult life. Focusing on a few smORF peptides whose functions have been elucidated recently, we discuss the importance of these newly identified regulatory molecules and how they act to regulate the building and function of the whole organism. PMID:26969979

  18. Changes in digestive enzyme activities during larval development of leopard grouper (Mycteroperca rosacea).

    PubMed

    Martínez-Lagos, R; Tovar-Ramírez, D; Gracia-López, V; Lazo, J P

    2014-06-01

    The leopard grouper is an endemic species of the Mexican Pacific with an important commercial fishery and good aquaculture potential. In order to assess the digestive capacity of this species during the larval period and aid in the formulation of adequate weaning diets, this study aimed to characterize the ontogeny of digestive enzymes during development of the digestive system. Digestive enzymes trypsin, chymotrypsin, acid protease, leucine-alanine peptidase, alkaline phosphatase, aminopeptidase N, lipase, amylase and maltase were quantified in larvae fed live prey and weaned onto a formulated microdiet at 31 days after hatching (DAH) and compared with fasting larvae. Enzyme activity for trypsin, lipase and amylase were detected before the opening of the mouth and the onset of exogenous feeding, indicating a precocious development of the digestive system that has been described in many fish species. The intracellular enzyme activity of leucine-alanine peptidase was high during the first days of development, with a tendency to decrease as larvae developed, reaching undetectable levels at the end of the experimental period. In contrast, activities of enzymes located in the intestinal brush border (i.e., aminopeptidase and alkaline phosphatase) were low at the start of exogenous feeding but progressively increased with larval development, indicating the gradual maturation of the digestive system. Based on our results, we conclude that leopard grouper larvae possess a functional digestive system at hatching and before the onset of exogenous feeding. The significant increase in the activity of trypsin, lipase, amylase and acid protease between 30 and 40 DAH suggests that larvae of this species can be successfully weaned onto microdiets during this period. PMID:24189829

  19. Anatomy and development of the larval nervous system in Echinococcus multilocularis

    PubMed Central

    2013-01-01

    Background The metacestode larva of Echinococcus multilocularis (Cestoda: Taeniidae) develops in the liver of intermediate hosts (typically rodents, or accidentally in humans) as a labyrinth of interconnected cysts that infiltrate the host tissue, causing the disease alveolar echinococcosis. Within the cysts, protoscoleces (the infective stage for the definitive canid host) arise by asexual multiplication. These consist of a scolex similar to that of the adult, invaginated within a small posterior body. Despite the importance of alveolar echinococcosis for human health, relatively little is known about the basic biology, anatomy and development of E. multilocularis larvae, particularly with regard to their nervous system. Results We describe the existence of a subtegumental nerve net in the metacestode cysts, which is immunoreactive for acetylated tubulin-α and contains small populations of nerve cells that are labeled by antibodies raised against several invertebrate neuropeptides. However, no evidence was found for the existence of cholinergic or serotoninergic elements in the cyst wall. Muscle fibers occur without any specific arrangement in the subtegumental layer, and accumulate during the invaginations of the cyst wall that form brood capsules, where protoscoleces develop. The nervous system of the protoscolex develops independently of that of the metacestode cyst, with an antero-posterior developmental gradient. The combination of antibodies against several nervous system markers resulted in a detailed description of the protoscolex nervous system, which is remarkably complex and already similar to that of the adult worm. Conclusions We provide evidence for the first time of the existence of a nervous system in the metacestode cyst wall, which is remarkable given the lack of motility of this larval stage, and the lack of serotoninergic and cholinergic elements. We propose that it could function as a neuroendocrine system, derived from the nervous system

  20. Effect of larval food amount on ovariole development in queens of Trigona spinipes (Hymenoptera, Apinae).

    PubMed

    Lisboa, L C O; Serrão, J E; Cruz-Landim, C; Campos, L A O

    2005-06-01

    Caste determination in Trigona spinipes Fabricius (Hymenoptera, Apidae, Meliponini) is trophogenic. Larvae that eat about 360 microl of food become queens, while those who consume 36 microl develop into workers. We studied the effect of larval nutrition on the number and length of ovarioles and on ovarian development in fifth instar larvae, white eyed, pink eyed and black-eyed pupae as well as newly emerged adults. All larvae have four ovarioles per ovary, while in queen pupae this number ranged from 8 to 15. Cyst formation, the cell death and other characteristics of ovary morphogenesis were the same regardless of the quantity of food consumed. These results are discussed in relation to caste differentiation in other bees. PMID:15929734

  1. Otolith Growth and macular Carbonic Anhydrase Reactivity in larval Fish after Development at simulated Microgravity

    NASA Astrophysics Data System (ADS)

    Baur, U.; Hilbig, R.; Anken, R.

    Otolith growth in terms of mineralisation mainly depends on the enzyme carbonic anhydrase (CA). CA is located in specialized, mitochondria-rich macular cells (ionocytes), which are involved in the endolymphatic ion exchange, and the enzyme is responsible for the provision of the pH-value necessary for otolithic calcium carbonate deposition. Since it has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish via a down-regulation of CA reactivity, we were prompted to elucidate whether (simulated) microgravity would possibly yield opposite effects. Therefore, larval siblings of cichlid fish (Oreochromis mossambicus) were housed in a submersed, two-dimensional clinostat (tube) during their development. Subsequently, the "physical capacity" (i.e., size) of the otoliths was measured, CA was histochemically demonstrated in ionocytes, and enzyme reactivity was determined densitometrically. The respective data will be communicated at the meeting. Acknowledgement: This work was financially supported by the German Aerospace Center (DLR) (FKZ: 50 WB 9997).

  2. The behavior of larval zebrafish reveals stressor-mediated anorexia during early vertebrate development

    PubMed Central

    De Marco, Rodrigo J.; Groneberg, Antonia H.; Yeh, Chen-Min; Treviño, Mario; Ryu, Soojin

    2014-01-01

    The relationship between stress and food consumption has been well documented in adults but less so in developing vertebrates. Here we demonstrate that an encounter with a stressor can suppress food consumption in larval zebrafish. Furthermore, we provide indication that food intake suppression cannot be accounted for by changes in locomotion, oxygen consumption and visual responses, as they remain unaffected after exposure to a potent stressor. We also show that feeding reoccurs when basal levels of cortisol (stress hormone in humans and teleosts) are re-established. The results present evidence that the onset of stress can switch off the drive for feeding very early in vertebrate development, and add a novel endpoint for analyses of metabolic and behavioral disorders in an organism suitable for high-throughput genetics and non-invasive brain imaging. PMID:25368561

  3. The behavior of larval zebrafish reveals stressor-mediated anorexia during early vertebrate development.

    PubMed

    De Marco, Rodrigo J; Groneberg, Antonia H; Yeh, Chen-Min; Treviño, Mario; Ryu, Soojin

    2014-01-01

    The relationship between stress and food consumption has been well documented in adults but less so in developing vertebrates. Here we demonstrate that an encounter with a stressor can suppress food consumption in larval zebrafish. Furthermore, we provide indication that food intake suppression cannot be accounted for by changes in locomotion, oxygen consumption and visual responses, as they remain unaffected after exposure to a potent stressor. We also show that feeding reoccurs when basal levels of cortisol (stress hormone in humans and teleosts) are re-established. The results present evidence that the onset of stress can switch off the drive for feeding very early in vertebrate development, and add a novel endpoint for analyses of metabolic and behavioral disorders in an organism suitable for high-throughput genetics and non-invasive brain imaging. PMID:25368561

  4. Embryonic, Larval, and Early Juvenile Development of the Tropical Sea Urchin, Salmacis sphaeroides (Echinodermata: Echinoidea)

    PubMed Central

    Rahman, M. Aminur; Yusoff, Fatimah Md.; Arshad, A.; Shamsudin, Mariana Nor; Amin, S. M. N.

    2012-01-01

    Salmacis sphaeroides (Linnaeus, 1758) is one of the regular echinoids, occuring in the warm Indo-West Pacific, including Johor Straits, between Malaysia and Singapore. In order to investigate the developmental basis of morphological changes in embryos and larvae, we documented the ontogeny of S. sphaeroides in laboratory condition. Gametes were obtained from adult individuals by 0.5 M KCl injection into the coelomic cavity. Fertilization rate at limited sperm concentration (10−5 dilution) was 96.6 ± 1.4% and the resulting embryos were reared at 24°C. First cleavage (2-cell), 4-cell, 8-cell, 16-cell, 32-cell, and multicell (Morulla) stages were achieved 01.12, 02.03, 02.28, 02.51, 03.12, and 03.32 h postfertilization. Ciliated blastulae with a mean length of 174.72 ± 4.43 μm hatched 08.45 h after sperm entry. The gastrulae formed 16.15 h postfertilization and the archenteron elongated constantly while ectodermal red-pigmented cells migrated synchronously to the apical plate. Pluteus larva started to feed unicellular algae in 2 d, grew continuously, and finally attained metamorphic competence in 35 d after fertilization. Metamorphosis took approximately 1 h 30 min from attachment to the complete resorption of larval tissues and the development of complete juvenile structure with adult spines, extended tubefeet and well-developed pedicellaria, the whole event of which usually took place within 1 d postsettlement. This study represents the first successful investigation on embryonic, larval, and early juvenile development of S. sphaeroides. The findings would greatly be helpful towards the understanding of ontogeny and life-history strategies, which will facilitate us to develop the breeding, seed production, and culture techniques of sea urchins in captive condition. PMID:23055824

  5. Embryonic, larval, and early juvenile development of the tropical sea urchin, Salmacis sphaeroides (Echinodermata: Echinoidea).

    PubMed

    Rahman, M Aminur; Yusoff, Fatimah Md; Arshad, A; Shamsudin, Mariana Nor; Amin, S M N

    2012-01-01

    Salmacis sphaeroides (Linnaeus, 1758) is one of the regular echinoids, occuring in the warm Indo-West Pacific, including Johor Straits, between Malaysia and Singapore. In order to investigate the developmental basis of morphological changes in embryos and larvae, we documented the ontogeny of S. sphaeroides in laboratory condition. Gametes were obtained from adult individuals by 0.5 M KCl injection into the coelomic cavity. Fertilization rate at limited sperm concentration (10(-5) dilution) was 96.6 ± 1.4% and the resulting embryos were reared at 24°C. First cleavage (2-cell), 4-cell, 8-cell, 16-cell, 32-cell, and multicell (Morulla) stages were achieved 01.12, 02.03, 02.28, 02.51, 03.12, and 03.32 h postfertilization. Ciliated blastulae with a mean length of 174.72 ± 4.43 μm hatched 08.45 h after sperm entry. The gastrulae formed 16.15 h postfertilization and the archenteron elongated constantly while ectodermal red-pigmented cells migrated synchronously to the apical plate. Pluteus larva started to feed unicellular algae in 2 d, grew continuously, and finally attained metamorphic competence in 35 d after fertilization. Metamorphosis took approximately 1 h 30 min from attachment to the complete resorption of larval tissues and the development of complete juvenile structure with adult spines, extended tubefeet and well-developed pedicellaria, the whole event of which usually took place within 1 d postsettlement. This study represents the first successful investigation on embryonic, larval, and early juvenile development of S. sphaeroides. The findings would greatly be helpful towards the understanding of ontogeny and life-history strategies, which will facilitate us to develop the breeding, seed production, and culture techniques of sea urchins in captive condition. PMID:23055824

  6. Activities of natural methyl farnesoids on pupariation and metamorphosis of Drosophila melanogaster

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methyl farnesoate (MF) and juvenile hormone (JH III), which respectively bind to the receptors USP and MET, and bisepoxy JH III (bisJHIII) were assessed for several activities during Drosophila larval development, and during prepupal development to eclosed adults. Dietary MF and JH III were similar...

  7. Proteomic Analysis of Oesophagostomum dentatum (Nematoda) during Larval Transition, and the Effects of Hydrolase Inhibitors on Development

    PubMed Central

    Ondrovics, Martina; Silbermayr, Katja; Mitreva, Makedonka; Young, Neil D.; Razzazi-Fazeli, Ebrahim; Gasser, Robin B.; Joachim, Anja

    2013-01-01

    In this study, in vitro drug testing was combined with proteomic and bioinformatic analyses to identify and characterize proteins involved in larval development of Oesophagostomum dentatum, an economically important parasitic nematode. Four hydrolase inhibitors ο-phenanthroline, sodium fluoride, iodoacetamide and 1,2-epoxy-3-(pnitrophenoxy)-propane (EPNP) significantly inhibited (≥90%) larval development. Comparison of the proteomic profiles of the development-inhibited larvae with those of uninhibited control larvae using two-dimensional gel electrophoresis, and subsequent MALDI-TOF mass spectrometric analysis identified a down-regulation of 12 proteins inferred to be involved in various larval developmental processes, including post-embryonic development and growth. Furthermore, three proteins (i.e. intermediate filament protein B, tropomyosin and peptidyl-prolyl cis-trans isomerase) inferred to be involved in the moulting process were down-regulated in moulting- and development-inhibited O. dentatum larvae. This first proteomic map of O. dentatum larvae provides insights in the protein profile of larval development in this parasitic nematode, and significantly improves our understanding of the fundamental biology of its development. The results and the approach used might assist in developing new interventions against parasitic nematodes by blocking or disrupting their key biological pathways. PMID:23717515

  8. Evaluation of growth and development of hatchery-reared larval black sea bass centropristis striata via image analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To improve the understanding of larval development and to refine hatchery production techniques for black sea bass Centropristis striata, digital photography and image analysis were employed to characterize growth and development of larvae from hatch through metamorphosis. Within 12 h after hatchin...

  9. Volumetric Measurements of Amnioserosa Cells in Developing Drosophila

    NASA Astrophysics Data System (ADS)

    Mashburn, David; Jayasinghe, Aroshan; Hutson, Shane

    2013-03-01

    The behavior of cells in tissue in developing Drosophila melanogaster has become increasingly clearer over the past few decades, in large part due to advances in imaging techniques, genetic markers, predictive modeling, and micromanipulation (notably laser microsurgery). We now know apical contractions in amnioserosa cells are a significant factor in large scale processes like germ band retraction and dorsal closure. Also, laser microsurgery induces cellular recoil that strongly mimics a 2D elastic sheet. Still, what we know about these processes comes entirely from the apical surface where the standard fluorescent markers like cadherin are located, but many open questions exist concerning the remaining ``dark'' portion of cells. Does cell volume remain constant during contraction or do cells leak? Also, what shape changes do cells undergo? Do they bulge, wedge, contract prismatically, or something else? By using a marker that labels the entire membrane of amnioserosa cells (Resille, 117) and adapting our watershed segmentation routines for 4D datasets, we have been able to quantify the entire volumetric region of cells in tissue through time and compare changes in apical area and volume. Preliminary results suggest a fairly constant volume over the course of a contraction cycle.

  10. Ocean warming ameliorates the negative effects of ocean acidification on Paracentrotus lividus larval development and settlement.

    PubMed

    García, Eliseba; Clemente, Sabrina; Hernández, José Carlos

    2015-09-01

    Ocean warming and acidification both impact marine ecosystems. All organisms have a limited body temperature range, outside of which they become functionally constrained. Beyond the absolute extremes of this range, they cannot survive. It is hypothesized that some stressors can present effects that interact with other environmental variables, such as ocean acidification (OA) that have the potential to narrow the thermal range where marine species are functional. An organism's response to ocean acidification can therefore be highly dependent on thermal conditions. This study evaluated the combined effects of predicted ocean warming conditions and acidification, on survival, development, and settlement, of the sea urchin Paracentrotus lividus. Nine combined treatments of temperature (19.0, 20.5 and 22.5 °C) and pH (8.1, 7.7 and 7.4 units) were carried out. All of the conditions tested were either within the current natural ranges of seawater pH and temperature or are within the ranges that have been predicted for the end of the century, in the sampling region (Canary Islands). Our results indicated that the negative effects of low pH on P. lividus larval development and settlement will be mitigated by a rise in seawater temperature, up to a thermotolerance threshold. Larval development and settlement performance of the sea urchin P. lividus was enhanced by a slight increase in temperature, even under lowered pH conditions. However, the species did show negative responses to the levels of ocean warming and acidification that have been predicted for the turn of the century. PMID:26275754

  11. Combined endosulfan and cypermethrin-induced toxicity to embryo-larval development of Rhinella arenarum.

    PubMed

    Svartz, Gabriela V; Aronzon, Carolina M; Pérez Coll, Cristina S

    2016-01-01

    The combined effects of two widely used pesticides, endosulfan and cypermethrin, on survival of embryo-larval development of the South American toad (Rhinella arenarum) were examined. The toxicity bioassays were performed according to the AMPHITOX test. Embryos and larvae were exposed to mixtures of these pesticides at equitoxic ratios from acute or chronic exposure to evaluate interaction effects. The results were analyzed using both Marking's additive index and combination index (CI)-isobologram methods. Acute (96-h) and intermediate (168-h) toxicity of endosulfan-cypermethrin mixtures remained almost constant for larvae and embryos, but when exposure duration was increased, there was a significant elevation in toxicity, obtaining chronic (240-h) no-observed-effect concentrations (NOEC) values of 0.045 and 0.16 mg/L for embryos and larvae, respectively. These are environmentally relevant concentrations that reflect a realistic risk of this pesticide mixture to this native amphibian species. The toxicity increment with the exposure duration was coincident with the central nervous system development on embryos reaching the larval period, the main target organ of these pesticides. The interactions of the pesticide mixtures at acute and chronic exposure were antagonistic for embryo development (CI > 1), and additive (CI = 1) for larvae, while chronic exposure interactions were synergistic (CI < 1) for both developmental periods. Data indicated that endosulfan-cypermethrin mixtures resulted in different interaction types depending on duration and developmental stage exposed. As a general pattern and considering conditions of overall developmental period and chronic exposure, this pesticide mixture usually applied in Argentine crop fields is synergistic with respect to toxicity for this native amphibian species. PMID:26914601

  12. [Embrionary and larval development of Lytechinus variegatus (Echinoidea: Toxopneustidae) in laboratory conditions at Isla de Margarita-Venezuela].

    PubMed

    Gómez, Olga; Gómez, Alfredo

    2005-12-01

    The sea urchin Lytechinus variegatus is a promissory species for aquaculture activities in tropical countries. In Venezuela, this species has some economical importance but their embryonic and larval development had not been studied. We collected specimens from seagrass beds in Margarita Island (Venezuela) and kept them in the laboratory, where they spawned naturally. With filtered sea water (temperature 28 degrees C, salinity 37 psu) and moderate aeration, the eggs and sperm were mixed (relation 1:100) and reached a 90% fertilization rate. The fertilization envelope was observed after two minutes, the first cellular division after 45 minutes and the prism larval stage after 13 hours. The echinopluteus larval stage was reached after 17 hours and metamorphosis after 18 days of planktonic life, when the larvae start their benthic phase. PMID:17469261

  13. Temperature, Larval Diet, and Density Effects on Development Rate and Survival of Aedes aegypti (Diptera: Culicidae)

    PubMed Central

    Couret, Jannelle; Dotson, Ellen; Benedict, Mark Q.

    2014-01-01

    Many environmental factors, biotic and abiotic interact to influence organismal development. Given the importance of Aedes aegypti as a vector of human pathogens including dengue and yellow fever, understanding the impact of environmental factors such as temperature, resource availability, and intraspecific competition during development is critical for population control purposes. Despite known associations between developmental traits and factors of diet and density, temperature has been considered the primary driver of development rate and survival. To determine the relative importance of these critical factors, wide gradients of conditions must be considered. We hypothesize that 1) diet and density, as well as temperature influence the variation in development rate and survival, 2) that these factors interact, and this interaction is also necessary to understand variation in developmental traits. Temperature, diet, density, and their two-way interactions are significant factors in explaining development rate variation of the larval stages of Ae. aegypti mosquitoes. These factors as well as two and three-way interactions are significantly associated with the development rate from hatch to emergence. Temperature, but not diet or density, significantly impacted juvenile mortality. Development time was heteroskedastic with the highest variation occurring at the extremes of diet and density conditions. All three factors significantly impacted survival curves of experimental larvae that died during development. Complex interactions may contribute to variation in development rate. To better predict variation in development rate and survival in Ae. aegypti, factors of resource availability and intraspecific density must be considered in addition, but never to the exclusion of temperature. PMID:24498328

  14. The formation of the nervous system during larval development in Triops cancriformis (Bosc) (crustacea, Branchiopoda): An immunohistochemical survey.

    PubMed

    Fritsch, Martin; Richter, Stefan

    2010-12-01

    We provide data of the development of thenervous system during the first five larval stages of Triops cancriformis. We use immunohistochemical labeling (against acetylated α-tubulin, serotonin, histamine, and FMRFamide), confocal laser scanning microscopy analysis, and 3D-reconstruction. The development of the nervous system corresponds with the general anamorphic development in T. cancriformis. In larval stage I (L I), all brain parts (proto-, deuto-, and tritocerebrum), the circumoral connectives, and the mandibular neuromere are already present. Also, the frontal filaments and the developing nauplius eye are already present. However, until stage L III, the nauplius eye only consists of three cups. Throughout larval development, the protocerebral network differentiates into distinct subdivisions. In the postnaupliar region, additional neuromeres and their commissures emerge in an anteroposterior gradient. The larval nervous system in L V consists of a differentiated protocerebrum including a central body, a nauplius eye comprising four cups, a circumoral nerve ring, mandibular- and postnaupliar neuromeres up to the seventh thoracic segment, each featuring an anterior and a posterior commissure, and two parallel connectives. The presence of a protocerebral bridge is questionable. The distribution of neurotransmitters in L I is restricted to the naupliar nervous system. Over the course of the five stages of development, neurotransmitter distribution also follows an anteroposterior gradient. Each neuromere is equipped with two ganglia innervating the locomotional appendages and possesses a specific neurotransmitter distribution pattern. We suggest a correlation between neurotransmitter expression and locomotion. PMID:20938985

  15. Exploration of the “larval pool”: development and ground-truthing of a larval transport model off leeward Hawai‘i

    PubMed Central

    Kobayashi, Donald R.

    2016-01-01

    Most adult reef fish show site fidelity thus dispersal is limited to the mobile larval stage of the fish, and effective management of such species requires an understanding of the patterns of larval dispersal. In this study, we assess larval reef fish distributions in the waters west of the Big Island of Hawai‘i using both in situ and model data. Catches from Cobb midwater trawls off west Hawai‘i show that reef fish larvae are most numerous in offshore waters deeper than 3,000 m and consist largely of pre-settlement Pomacanthids, Acanthurids and Chaetodontids. Utilizing a Lagrangian larval dispersal model, we were able to replicate the observed shore fish distributions from the trawl data and we identified the 100 m depth strata as the most likely depth of occupancy. Additionally, our model showed that for larval shore fish with a pelagic larval duration longer than 40 days there was no significant change in settlement success in our model. By creating a general additive model (GAM) incorporating lunar phase and angle we were able to explain 67.5% of the variance between modeled and in situ Acanthurid abundances. We took steps towards creating a predictive larval distribution model that will greatly aid in understanding the spatiotemporal nature of the larval pool in west Hawai‘i, and the dispersal of larvae throughout the Hawaiian archipelago. PMID:26855873

  16. Fine-scale temperature fluctuation and modulation of Dirofilaria immitis larval development in Aedes aegypti.

    PubMed

    Ledesma, Nicholas; Harrington, Laura

    2015-04-15

    We evaluated degree-day predictions of Dirofilaria immitis development (HDU) under constant and fluctuating temperature treatments of equal average daily temperature. Aedes aegypti mosquitoes were infected with D. immitis microfilariae and parasite development was recorded at set time points in dissected mosquitoes. Time to L3 development in Malpighian tubules and detection in mosquito heads was shorter for larvae experiencing a daily regime of 19±9°C than larvae at constant 19°C; larval development rate in Malpighian tubules was slower in fluctuating regimes maintained above the 14°C developmental threshold than larvae under constant temperatures. We showed that hourly temperature modeling more accurately predicted D. immitis development to infective L3 stage. Development time differed between fluctuating and constant temperature treatments spanning the 14°C development threshold, implicating a physiological basis for these discrepancies. We conclude that average daily temperature models underestimate L3 development-and consequently dog heartworm transmission risk-at colder temperatures, and spatiotemporal models of D. immitis transmission risk should use hourly temperature data when analyzing high daily temperature ranges spanning 14°C. PMID:25747489

  17. GROWTH AND CHANGES IN BIOCHEMICAL COMPOSITION DURING LARVAL DEVELOPMENT OF THE STONE CRAB, MENIPPE ADINA WILLIAMS AND FELDER, 1986

    EPA Science Inventory

    Larval development in Menippe adina was associated with changes in weight and biochemical composition. Larvae of the stone crab, M. adina, were mass-reared under laboratory conditions (28|C; 20o/ooS) from hatching to the megalopal stage. Growth in M. adina is exponential througho...

  18. Oviposition and larval development of a stem borer, Eoreuma loftini, on rice and non-crop grass hosts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A greenhouse study compared oviposition preference and larval development duration of a stem borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae), on rice, Oryza sativa L. (cv. Cocodrie), and four primary non-crop hosts of Gulf Coast Texas rice agroecosystems. Rice and two perennials, johnsongrass...

  19. Embryonic and larval development in the caecilian Ichthyophis kohtaoensis (Amphibia, gymnophiona): a staging table.

    PubMed

    Dünker, N; Wake, M H; Olson, W M

    2000-01-01

    Little is known about the developmental biology of caecilians-tropical, elongate, limbless, mostly fossorial amphibians that are members of the Order Gymnophiona. Ichthyophis kohtaoensis (Family Ichthyophiidae; southeast Asia) is an oviparous species in which maternal care of the clutch is provided. The clutch is laid in a burrow on land, and the embryos develop in their egg membranes, curved around a large yolk mass. Larvae are aquatic and exhibit characteristic features that are not present in the terrestrial adults. Because accurate descriptions of ontogenies and the establishment of standardized stages of embryonic and larval development are useful for both experimental and comparative embryology, a staging table for I.kohtaoensis was developed based on external morphological features. Development from the end of neurulation to metamorphosis was divided into 20 stages. Principal diagnostic features include development of the lateral line organs, formation of three pairs of external gills, development of the eyes, changes in yolk structure, changes in the structure of the cloacal aperture and growth of the tail, including the formation and regression of the tail fin. This study provides a comparison with descriptions of embryonic stages of I.glutinosus and Hypogeophis rostratus and with a recent staging table for the aquatic, viviparous caecilian Typhlonectes compressicauda, the only other caecilians for which reasonably complete ontogenetic information exists in the literature. Comparisons with established staging tables for selected frogs and salamanders are also presented. PMID:10629095

  20. Microarray Analysis of Juvenile Hormone Response in Drosophila melanogaster S2 cells

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A microchip array encompassing probes for 14,010 genes of Drosophila melanogaster was used to analyze the effect of juvenile hormone (JH) on genome-wide gene expression. JH is a member of a key group of insect hormones involved in regulating larval development and adult reproductive processes. Altho...

  1. EGFR Signaling in the Brain Is Necessary for Olfactory Learning in "Drosophila" Larvae

    ERIC Educational Resources Information Center

    Rahn, Tasja; Leippe, Matthias; Roeder, Thomas; Fedders, Henning

    2013-01-01

    Signaling via the epidermal growth factor receptor (EGFR) pathway has emerged as one of the key mechanisms in the development of the central nervous system in "Drosophila melanogaster." By contrast, little is known about the functions of EGFR signaling in the differentiated larval brain. Here, promoter-reporter lines of EGFR and its most prominent…

  2. Control of imaginal cell development by the escargot gene of Drosophila.

    PubMed

    Hayashi, S; Hirose, S; Metcalfe, T; Shirras, A D

    1993-05-01

    Mutations in the escargot (esg) locus, which codes for a zinc-finger-containing protein with similarity to the product of the snail gene, cause a variety of defects in adult structures such as loss of abdominal cuticle and malformation of the wings and legs. esg RNA is expressed in wing, haltere, leg and genital imaginal discs and in abdominal histoblast nests in the embryo. Expression in imaginal tissues is also found in third instar larvae. In esg mutant larvae, normally diploid abdominal histoblasts replicate their DNA without cell division and become similar in appearance to the polytene larval epidermal cells. A similar phenotype was also found in imaginal discs of larvae mutant for both esg and the Drosophila raf gene. These results suggest that one of the normal functions of esg may be the maintenance of diploidy in imaginal cells. PMID:8375329

  3. Ontogeny and phylogeny: molecular signatures of selection, constraint, and temporal pleiotropy in the development of Drosophila

    PubMed Central

    Artieri, Carlo G; Haerty, Wilfried; Singh, Rama S

    2009-01-01

    Background Karl Ernst Von Baer noted that species tend to show greater morphological divergence in later stages of development when compared to earlier stages. Darwin originally interpreted these observations via a selectionist framework, suggesting that divergence should be greatest during ontogenic stages in which organisms experienced varying 'conditions of existence' and opportunity for differential selection. Modern hypotheses have focused on the notion that genes and structures involved in early development will be under stronger purifying selection due to the deleterious pleiotropic effects of mutations propagating over the course of ontogeny, also known as the developmental constraint hypothesis. Results Using developmental stage-specific expressed sequence tag (EST) libraries, we tested the 2 hypotheses by comparing the rates of evolution of 7,180 genes obtained from 6 species of the Drosophila melanogaster group with respect to ontogeny, and sex and reproduction-related functions in gonadal tissues. Supporting morphological observations, we found evidence of a pattern of increasing mean evolutionary rate in genes that are expressed in subsequent stages of development. Furthermore, supporting expectations that early expressed genes are constrained in divergence, we found that embryo stage genes are involved in a higher mean number of interactions as compared to later stages. We noted that the accelerated divergence of genes in the adult stage is explained by those expressed specifically in the male gonads, whose divergence is driven by positive selection. In addition, accelerated gonadal gene divergence occurs only in the adult stage, suggesting that the effects of selection are observed primarily at the stages during which they are expected occur. Finally, we also found a significant correlation between temporal specificity of gene expression and evolutionary rate, supporting expectations that genes with ubiquitous expression are under stronger constraint

  4. Effects of temperature and salinity on larval development of Elminius modestus (Crustacea, Cirripedia) from Helgoland (North Sea) and New Zealand

    NASA Astrophysics Data System (ADS)

    Harms, J.

    1986-12-01

    Larvae of Elminius modestus (Darwin) from four different populations (Portobello, Leigh, Doubtless Bay [New Zealand] and Helgoland [North Sea]) were reared at different salinity and temperature combinations. The larvae of E. modestus from Helgoland developed successfully at a wide range of temperature (6° to 24 °C) and salinity (20 to 50 ‰ S). Mortality was highest at 10 ‰ S; only at 12° and 18 °C did a small percentage develop to the cypris. The larvae from New Zealand were reared at a temperature range of 12° 24 °C at 20, 30 and 40 ‰ S; mortality increased in all populations at all salinities with decreasing temperature and was extremely high at 12 °C and 40 ‰ S. The temperature influence on larval duration could be described in all cases by a power function. No significant differences in temperature influences on developmental times between the tested salinities were found, except for the Portobello population at 20 ‰ S. Significant differences were found in the temperature influence on larval development between the populations from Helgoland and the North Island of New Zealand (Leigh, Doubtless Bay). No differences were found between the Helgoland and Portobello population. The pooled data for the temperature influence on the larval development of the three tested New Zealand populations at 20, 30 and 40 ‰ S and the pooled Helgoland data at 20, 30 and 40 ‰ S show highly significant differences. Larval size (stage VI) was influenced by experimental conditions. The larvae grew bigger at low temperatures and attained their maximum size at 30 ‰ S (Helgoland). There was a strong reduction in larval size at temperatures from 18° to 24 °C. The larvae of the New Zealand populations were smaller than those from Helgoland. The greatest difference in size existed between the larvae from Portobello and Helgoland.

  5. Evolution and plasticity of anuran larval development in response to desiccation. A comparative analysis.

    PubMed

    Richter-Boix, Alex; Tejedo, Miguel; Rezende, Enrico L

    2011-09-01

    Anurans breed in a variety of aquatic habitats with contrasting levels of desiccation risk, which may result in selection for faster development during larval stages. Previous studies suggest that species in ephemeral ponds reduce their developmental times to minimize desiccation risks, although it is not clear how variation in desiccation risk affects developmental strategies in different species. Employing a comparative phylogenetic approach including data from published and unpublished studies encompassing 62 observations across 30 species, we tested if species breeding in ephemeral ponds (High risk) develop faster than those from permanent ponds (Low risk) and/or show increased developmental plasticity in response to drying conditions. Our analyses support shorter developmental times in High risk, primarily by decreasing body mass at metamorphosis. Plasticity in developmental times was small and did not differ between groups. However, accelerated development in High risk species generally resulted in reduced sizes at metamorphosis, while some Low risk species were able compensate this effect by increasing mean growth rates. Taken together, our results suggest that plastic responses in species breeding in ephemeral ponds are constrained by a general trade-off between development and growth rates. PMID:22393479

  6. Developing a Novel Embryo-Larval Zebrafish Xenograft Assay to Prioritize Human Glioblastoma Therapeutics.

    PubMed

    Wehmas, Leah Christine; Tanguay, Robert L; Punnoose, Alex; Greenwood, Juliet A

    2016-08-01

    Glioblastoma is an aggressive brain cancer requiring improved treatments. Existing methods of drug discovery and development require years before new therapeutics become available to patients. Zebrafish xenograft models hold promise for prioritizing drug development. We have developed an embryo-larval zebrafish xenograft assay in which cancer cells are implanted in a brain microenvironment to discover and prioritize compounds that impact glioblastoma proliferation, migration, and invasion. We illustrate the utility of our assay by evaluating the well-studied, phosphatidylinositide 3-kinase inhibitor LY294002 and zinc oxide nanoparticles (ZnO NPs), which demonstrate selective cancer cytotoxicity in cell culture, but the in vivo effectiveness has not been established. Exposures of 3.125-6.25 μM LY294002 significantly decreased proliferation up to 34% with concentration-dependent trends. Exposure to 6.25 μM LY294002 significantly inhibited migration/invasion by ∼27% within the glioblastoma cell mass (0-80 μm) and by ∼32% in the next distance region (81-160 μm). Unexpectedly, ZnO enhanced glioblastoma proliferation by ∼19% and migration/invasion by ∼35% at the periphery of the cell mass (161+ μm); however, dissolution of these NPs make it difficult to discern whether this was a nano or ionic effect. These results demonstrate that we have a short, relevant, and sensitive zebrafish-based assay to aid glioblastoma therapeutic development. PMID:27158859

  7. Evolution of poecilogony from planktotrophy: cryptic speciation, phylogeography, and larval development in the gastropod genus Alderia.

    PubMed

    Ellingson, Ryan A; Krug, Patrick J

    2006-11-01

    Poecilogony, a rare phenomenon in marine invertebrates, occurs when alternative larval morphs differing in dispersal potential or trophic mode are produced from a single genome. Because both poecilogony and cryptic species are prevalent among sea slugs in the suborder Sacoglossa (Gastropoda: Opisthobranchia), molecular data are needed to confirm cases of variable development and to place them in a phylogenetic context. The nominal species Alderia modesta produces long-lived, feeding larvae throughout the North Atlantic and Pacific, but in California can also produce short-lived larvae that metamorphose without feeding. We collected morphological, developmental, and molecular data for Alderia from 17 sites spanning the eastern and western Pacific and North Atlantic. Estuaries south of Bodega Harbor, California, contained a cryptic species (hereafter Alderia sp.) with variable development, sister to the strictly planktotrophic A. modesta. The smaller Alderia sp. seasonally toggled between planktotrophy and lecithotrophy, with some individuals differing in development but sharing mitochondrial DNA haplotypes. The sibling species overlapped in Tomales Bay, California, but showed no evidence of hybridization; laboratory mating trials suggest postzygotic isolation has arisen. Intra- and interspecific divergence times were estimated using a molecular clock calibrated with geminate sacoglossans. Speciation occurred about 4.1 million years ago during a major marine radiation in the eastern Pacific, when large inland embayments in California may have isolated ancestral populations. Atlantic and Pacific A. modesta diverged about 1.7 million years ago, suggesting trans-Arctic gene flow was interrupted by Pleistocene glaciation. Both Alderia species showed evidence of late Pleistocene population expansion, but the southern Alderia sp. likely experienced a more pronounced bottleneck. Reduced body size may have incurred selection against obligate planktotrophy in Alderia sp. by

  8. INFLUENCE OF AN INSECT GROWTH REGULATOR ON LARVAL DEVELOPMENT OF A MARINE CRUSTACEAN

    EPA Science Inventory

    Larval survival, growth, and energy metabolism of an estuarine shrimp (Palaemonetes pugio) were altered by exposure to low micrograms/l concentrations of an insect growth regulator (the juvenile hormone analogue, methoprene). Larvae were several orders of magnitude more sensitive...

  9. VARIATION IN GROWTH, LIPID CLASS AND FATTY ACID COMPOSITION OF THE MUD CRAB, RHITHROPANOPEUS HARRISII (GOULD) DURING LARVAL DEVELOPMENT FOLLOWING EXPOSURE TO AN INSECT JUVENILE HORMONE ANALOG (FENOXYCARB)

    EPA Science Inventory

    This study examines the effects of fenoxycarb?, an insect juvenile hormone analog, on larval growth, and lipid class and fatty acid composition in first crabs of the mud crab Rhithropanopeus harrisii reared through total larval development in nominal water concentrations from 1 ...

  10. Larval serum proteins of the gypsy moth, Lymantria dispar: Allometric changes during development suggest several functions for arylphorin and lipophorin

    SciTech Connect

    Karpells, S.T.

    1989-01-01

    Storage proteins are the major nutritive intermediates in insects and although the serum storage proteins are relatively well studied, definitive roles for many of them have yet to be established. To further characterize their roles in development and to establish quantitative baselines for future studies, two serum proteins, arylphorin (Ap) and lipophorin (Lp), of the gypsy moth, Lymantria dispar, were studied. Ap and Lp, isolated from larval hemolymph, were partially characterized biochemically and immunologically. Hemolymph concentrations throughout larval development were determined using quantitative immunoelectrophoresis and absolute hemolymph amounts of protein were determined by measuring hemolymph volume. Cyclic fluctuations in hemolymph concentrations of Ap in particular correlated with each molting cycle and an increase in Lp levels just prior to pupation suggest a metamorphic change in the role or demand for the protein. Sexual dimorphism in protein concentrations are explained in part by the sexual dimorphism in the number of larval instars. In fact, an additional instar of Ap accumulation in the female gypsy moth is suggested to compensate for the lack of a female-specific storage protein in this species. The last two days of each instar were found to be the optimum time to sample protein concentration with minimum variance. Allometric relationships among Ap accumulation, Lp accumulation and weight gain were uncovered. Ap labelled with ({sup 14}C)-N-ethylmaleimide was shown to be incorporated into newly synthesized cuticle and setae during a larval-larval molt. The antiserum developed against L. dispar Ap was used to identify the Ap of Trichoplusia in and study Ap titers in parasitized T. in larvae. The antiserum was also used to determine the immunological relatedness of 5 species of Lepidoptera.

  11. Influence of starvation on the larval development of Hyas araneus (Decapoda, Majidae)

    NASA Astrophysics Data System (ADS)

    Anger, K.; Dawirs, R. R.

    1981-09-01

    The influence of starvation on larval development of the spider crab Hyas araneus (L.) was studied in laboratory experiments. No larval stage suffering from continual lack of food had sufficient energy reserves to reach the next instar. Maximal survival times were observed at four different constant temperatures (2°, 6°, 12° and 18 °C). In general, starvation resistance decreased as temperatures increased: from 72 to 12days in the zoea-1, from 48 to 18 days in the zoea-2, and from 48 to 15 days in the megalopa stage. The length of maximal survival is of the same order of magnitude as the duration of each instar at a given temperature. “Sublethal limits” of early starvation periods were investigated at 12 °C: Zoea larvae must feed right from the beginning of their stage (at high food concentration) and for more than one fifth, approximately, of that stage to have at least some chance of surviving to the next instar, independent of further prey availability. The minimum time in which enough reserves are accumulated for successfully completing the instar without food is called “point-of-reserve-saturation” (PRS). If only this minimum period of essential initial feeding precedes starvation, development in both zoeal stages is delayed and mortality is greater, when compared to the fed control. Starvation periods beginning right after hatching of the first zoea cause a prolongation of this instar and, surprisingly, a slight shortening of the second stage. The delay in the zoea-1 increases proportionally to the length of the initial fasting period. If more than approximately 70 % of the maximum possible survival time has elapsed without food supply, the larvae become unable to recover and to moult to the second stage even when re-fed (“point-of-no-return”, PNR). The conclusion, based on own observations and on literature data, is that initial feeding is of paramount importance in the early development of planktotrophic decapod larvae. Taking into account

  12. 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

  13. 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

  14. Larval growth, development, and survival of laboratory-reared Aplysia californica: Effects of diet and veliger density*

    PubMed Central

    Capo, Thomas R.; Bardales, Ana T.; Gillette, Phillip R.; Lara, Monica R.; Schmale, Michael C.; Serafy, Joseph E.

    2009-01-01

    Over the last three decades, the California sea hare, Aplysia californica, has played an increasingly important role as a model organism in the neurosciences. Since 1995, the National Resource for Aplysia has supported a growing research community by providing a consistent supply of laboratory-reared individuals of known age, reproductive status, and environmental history. The purpose of the present study was to resolve the key biological factors necessary for successful culture of large numbers of high quality larval Aplysia. Data from a sequence of five experiments demonstrated that algal diet, food concentration, and veliger density significantly affected growth, attainment of metamorphic competency, and survival of Aplysia larvae. The highest growth and survival were achieved with a mixed algal diet of 1:1 Isochrysis sp (TISO) and Chaetoceros muelleri (CHGRA) at a total concentration of 250 x 103 cells/mL and a larval density of 0.5 – 1.0 per mL. Rapid growth was always correlated with faster attainment of developmental milestones and increased survival, indicating that the more rapidly growing larvae were healthier. Trials conducted with our improved protocol resulted in larval growth rates of >14 μm/d, which yielded metamorphically competent animals within 21 days with survival rates in excess of 90%. These data indicate the important effects of biotic factors on the critical larval growth period in the laboratory and show the advantages of developing optimized protocols for culture of such marine invertebrates. PMID:19000779

  15. The Drosophila neural lineages: a model system to study brain development and circuitry.

    PubMed

    Spindler, Shana R; Hartenstein, Volker

    2010-06-01

    In Drosophila, neurons of the central nervous system are grouped into units called lineages. Each lineage contains cells derived from a single neuroblast. Due to its clonal nature, the Drosophila brain is a valuable model system to study neuron development and circuit formation. To better understand the mechanisms underlying brain development, genetic manipulation tools can be utilized within lineages to visualize, knock down, or over-express proteins. Here, we will introduce the formation and development of lineages, discuss how one can utilize this model system, offer a comprehensive list of known lineages and their respective markers, and then briefly review studies that have utilized Drosophila neural lineages with a look at how this model system can benefit future endeavors. PMID:20306203

  16. The Drosophila neural lineages: a model system to study brain development and circuitry

    PubMed Central

    Spindler, Shana R.

    2010-01-01

    In Drosophila, neurons of the central nervous system are grouped into units called lineages. Each lineage contains cells derived from a single neuroblast. Due to its clonal nature, the Drosophila brain is a valuable model system to study neuron development and circuit formation. To better understand the mechanisms underlying brain development, genetic manipulation tools can be utilized within lineages to visualize, knock down, or over-express proteins. Here, we will introduce the formation and development of lineages, discuss how one can utilize this model system, offer a comprehensive list of known lineages and their respective markers, and then briefly review studies that have utilized Drosophila neural lineages with a look at how this model system can benefit future endeavors. PMID:20306203

  17. Embryonic and larval development in barfin flounder Verasper moseri (Jordan and Gilbert)

    NASA Astrophysics Data System (ADS)

    Du, Rongbin; Wang, Yongqiang; Jiang, Haibin; Liu, Liming; Wang, Maojian; Li, Tianbao; Zhang, Shubao

    2010-01-01

    Broodstock of Verasper moseri (Jordan and Gilbert) aged 3-4 years old were selected, and reinforced cultivation was conducted to promote maturation under controlled water temperature and photoperiod conditions. Fertilized eggs were obtained by artificial fertilization, and the development of embryos, larvae and juveniles was observed continuously. The results showed that the fertilized eggs of V. moseri were spherical, with transparent yolk and homogeneous bioplasm, and had no oil globule inside. The average diameter of the eggs was 1.77±0.02 mm. The eggs of V. moseri were buoyant in water with salinity above 35. The cleavage type was typical discoidal. Young pigment cells appeared when olfactory plates began to form. Hatching occurred at 187 h after fertilization at a water temperature of 8.5°C. The newly hatched larvae, floating on the water surface, were transparent with an average total length of 4.69±0.15 mm. During the cultivation period, when the water temperature was raised from 9 to 14.5°C, 4-day old larvae showed more melanophores on the body surface, making the larvae gray in color. The pectoral fins began to develop, which enabled the larvae to swim horizontally and in a lively manner. On days 7-8, the digestive duct formed. The yolk sac was small and black. The yolk sac was absorbed on day 11. Larvae took food actively, and body length and body height clearly increased. The rudiments of dorsal and anal fin pterygiophores were discernible and caudal fin ray elements formed on day 19. On day 24, the larval notochord flexed upwards, and the rays of unpaired fins began to differentiate. Pigment cells converged on the dorsal and anal fin rays, and the mastoid teeth on the mandible appeared. On day 29, the left eyes of juveniles began to move upwards. Depigmentation began in some juveniles and they became sandy brown in color on day 37. Most juveniles began to settle on the bottom of the tank. The left eyes of juveniles migrated completely to the right

  18. Calmodulin Point Mutations Affect Drosophila Development and Behavior

    PubMed Central

    Nelson, H. B.; Heiman, R. G.; Bolduc, C.; Kovalick, G. E.; Whitley, P.; Stern, M.; Beckingham, K.

    1997-01-01

    Calmodulin (CAM) is recognized as a major intermediary in intracellular calcium signaling, but as yet little is known of its role in developmental and behavioral processes. We have generated and studied mutations to the endogenous Cam gene of Drosophila melanogaster that change single amino acids within the protein coding region. One of these mutations produces a striking pupal lethal phenotype involving failure of head eversion. Various mutant combinations produce specific patterns of ectopic wing vein formation or melanotic scabs on the cuticle. Anaphase chromosome bridging is also seen as a maternal effect during the early embryonic nuclear divisions. In addition, specific behavioral defects such as poor climbing and flightlessness are detected among these mutants. Comparisons with other Drosophila mutant phenotypes suggests potential CAM targets that may mediate these developmental and behavioral effects, and analysis of the CAM crystal structure suggests the structural consequences of the individual mutations. PMID:9409836

  19. 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

  20. 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

  1. CF2 represses Actin 88F gene expression and maintains filament balance during indirect flight muscle development in Drosophila.

    PubMed

    Gajewski, Kathleen M; Schulz, Robert A

    2010-01-01

    The zinc finger protein CF2 is a characterized activator of muscle structural genes in the body wall muscles of the Drosophila larva. To investigate the function of CF2 in the indirect flight muscle (IFM), we examined the phenotypes of flies bearing five homozygous viable mutations. The gross structure of the IFM was not affected, but the stronger hypomorphic alleles caused an increase of up to 1.5X in the diameter of the myofibrils. This size increase did not cause any disruption of the hexameric arrangement of thick and thin filaments. RT-PCR analysis revealed an increase in the transcription of several structural genes. Ectopic overexpression of CF2 in the developing IFM disrupts muscle formation. While our results indicate a role for CF2 as a direct negative regulator of the thin filament protein gene Actin 88F (Act88F), effects on levels of transcripts of myosin heavy chain (mhc) appear to be indirect. This role is in direct contrast to that described in the larval muscles, where CF2 activates structural gene expression. The variation in myofibril phenotypes of CF2 mutants suggest the CF2 may have separate functions in fine-tuning expression of structural genes to insure proper filament stoichiometry, and monitoring and/or controlling the final myofibril size. PMID:20520827

  2. Role of the Deubiquitylating Enzyme DmUsp5 in Coupling Ubiquitin Equilibrium to Development and Apoptosis in Drosophila melanogaster

    PubMed Central

    Kovács, Levente; Nagy, Olga; Pál, Margit; Udvardy, Andor; Popescu, Octavian; Deák, Péter

    2015-01-01

    Protein ubiquitylation is a dynamic process that affects the function and stability of proteins and controls essential cellular processes ranging from cell proliferation to cell death. This process is regulated through the balanced action of E3 ubiquitin ligases and deubiquitylating enzymes (DUB) which conjugate ubiquitins to, and remove them from target proteins, respectively. Our genetic analysis has revealed that the deubiquitylating enzyme DmUsp5 is required for maintenance of the ubiquitin equilibrium, cell survival and normal development in Drosophila. Loss of the DmUsp5 function leads to late larval lethality accompanied by the induction of apoptosis. Detailed analyses at a cellular level demonstrated that DmUsp5 mutants carry multiple abnormalities, including a drop in the free monoubiquitin level, the excessive accumulation of free polyubiquitins, polyubiquitylated proteins and subunits of the 26S proteasome. A shortage in free ubiquitins results in the induction of a ubiquitin stress response previously described only in the unicellular budding yeast. It is characterized by the induction of the proteasome-associated deubiquitylase DmUsp14 and sensitivity to cycloheximide. Removal of DmUsp5 also activates the pro-apoptotic machinery thereby resulting in widespread apoptosis, indicative of an anti-apoptotic role of DmUsp5. Collectively, the pleiotropic effects of a loss of DmUsp5 function can be explained in terms of the existence of a limited pool of free monoubiquitins which makes the ubiquitin-dependent processes mutually interdependent. PMID:25806519

  3. Expression profile of the cadherin family in the developing Drosophila brain.

    PubMed

    Fung, Siaumin; Wang, Fay; Chase, Maretta; Godt, Dorothea; Hartenstein, Volker

    2008-01-20

    The Drosophila genome encodes 17 members of the cadherin family of adhesion molecules, which in vertebrates has been implicated in patterning the nervous system through cell and axon sorting. With only a few exceptions all cadherins show widespread expression in the larval brain. What expression patterns have in common is that 1) they are global, in the sense that all lineages of the central brain or optic lobe, or both, show expression; and 2) expression is stage-specific: some cadherins are expressed only in primary neurons (located closest to the neuropile), others in early secondary neurons (near the brain surface), or primaries plus late secondaries. The Fat-like cadherins, Fat and Dachsous, as well as Cad96Ca and Cad74A, are expressed in the epithelial optic lobe anlagen, which matches the widespread epithelial expression of these molecules in the embryo. DE-cadherin is restricted to immature secondary neurons and glia; by contrast, DN-cadherin, Flamingo, Cad87A, Cad99C, and Calsyntenin-1 appear in differentiating primary neurons and, at a later stage, some or all secondary neurons. Cad87A is strongly enriched apically in epithelia and in neuronal dendrites. Fat-like, Cad86C, Cad88C, Cad89D, and Dret are expressed ubiquitously in embryonic and larval brains at low or moderate levels. We conclude from this analysis that cadherins are likely to play a role in 'generic' neural functions, such as neurite fasciculation, branching, and synapse formation. PMID:18041774

  4. Laboratory observation on spawning, fecundity and larval development of amphioxus ( Branchiostoma belcheri Tsingtaunese)

    NASA Astrophysics Data System (ADS)

    Wu, Xian-Han; Zhang, Shi-Cui; Wang, Yong-Yuan; Zhang, Bao-Lu; Qu, Yan-Mei; Jiang, Xin-Ji

    1994-12-01

    Although amphioxus is widespread in temperate and tropical seas, its population is diminishing because of environmental pollution. To keep the population of this evolutionarily important animal from diminishing, study on its reproduction and development is necessary. The main findings in this study on the spawning and fecundity of the amphioxus reared in laboratory and its larval development are as follows. 1. Water temperature markedly affected the spawning. It spawned only when water temperature reached 21°C. 2. Spawning of the amphioxus in laboratory was markedly extended. Initially, the amphioxus spawned at about 7:00 PM, but spawning time was postponed as spawning days went on. 3. The number of eggs produced by a female ranged from 1400 to 12800, average of 5800. This also represents the fecundity of the amphioxus because it shedded all eggs within the ovary at a time. 4. During the first few months of life of the amphioxus, its growth rate changed seasonally. The growth rate in summer and fall was greater than that in winter. 5. The pelagic larva became a benthic adult after 50 days. 6. The amphioxus reared in laboratory from fertilized eggs could produce fertile eggs and sperms. These findings can be a foundation for measures to address the problem of diminishing amphioxus population.

  5. Acidic intracellular pH shift during Caenorhabditis elegans larval development

    SciTech Connect

    Wadsworth, W.G.; Riddle, D.L. )

    1988-11-01

    During recovery from the developmentally arrested, nonfeeding dauer stage of the nemotode Caenorhabditis elegans, metabolic activation is accompanied by a decrease in intracellular pH (pH{sub i}). Phosphorus-31 nuclear magnetic resonance ({sup 31}P NMR) analyses of perchloric acid extracts show that inorganic phosphate predominates in dauer larvae, whereas ATP and other high-energy metabolites are abundant within 6 hr after dauer larvae have been placed in food to initiate development. Although metabolic activation has been associated with an alkaline pH{sub i} shift in other organisms, in vivo {sup 31}P NMR analysis of recovering dauer larvae shows a pH{sub i} decrease from {approx} 7.3 to {approx} 6.3 within 3 hr after the animals encounter food. This shift occurs before feeding begins, and it coincides with, or soon follows, the developmental commitment to recover from the dauer stage, suggesting that control of pH{sub i} may be important in the regulation of larval development in nematodes.

  6. The development of the digestive tract in larval European catfish (Silurus glanis L.).

    PubMed

    Kozarić, Z; Kuzir, S; Petrinec, Z; Gjurcević, E; Bozić, M

    2008-04-01

    The European catfish, Silurus glanis L., has become an important aqua cultural fish in Croatia, and it is cultivated extensively in ponds in polyculture with carps. The development of the digestive tract in S. glanis was studied with the aim of improving intensive fish production. Research was carried out on S. glanis larval stadium from 1- to 19-day post-hatching (DPH). The main histological methods used were: haematoxylin and eosin staining, periodic acid Schiff staining (PAS), Alcian blue (AB) and toluidin blue staining (TB). A yolk sac was present during the first 5 days (1-5-DPH). During the initial 3-DPH period, there was no trace PAS and AB activity in the digestive tract. Differentiation of the digestive tract began at 3- to 5-DPH. The oesophagus was positive for AB at 5-DPH, PAS and TB after 7-DPH. Differentiation of enterocytes began at 5-DPG and the intestines were complete at 11-DPH. Development of liver and pancreas was also studied. The analysis of data obtained in this study suggests that after 5-DPH catfish larvae have morphologically completed digestive tracts. PMID:18333856

  7. Early embryo and larval development of inviable intergeneric hybrids derived from Crassostrea angulata and Saccostrea cucullata

    NASA Astrophysics Data System (ADS)

    Su, Jiaqi; Wang, Zhaoping; Zhang, Yuehuan; Yan, Xiwu; Li, Qiongzhen; Yu, Ruihai

    2016-06-01

    To detect the intergeneric hybridization between the oyster Crassostrea angulata and Saccostrea cucullata coexisting along the southern coast of China, reciprocal crosses were conducted between the two species. Barriers for sperm recognizing, binding, penetrating the egg, and forming the pronucleus were detected by fluorescence staining. From the results, although fertilization success was observed in hybrid crosses, the overall fertilization rate was lower than that of intraspecific crosses. A large number of hybrid larvae died at 6-8 d after hatching, and those survived could not complete metamorphosis. C. angulata ♀× S. cucullata ♂ larvae had a growth rate similar to that of the maternal species, whereas S. cucullata ♀ × C. angulata ♂ larvae grew the slowest among all crosses. Molecular genetics analysis revealed that hybrid progeny were amphimixis hybrids. This study demonstrated that hybrid embryos generated by crossing C. angulata and S. cucullata could develop normally to the larval state, but could not complete metamorphosis and then develop to the spat stage. Thus, there is a post-reproductive isolation between C. angulata and S. cucullata.

  8. Development of the GABA-ergic signaling system and its role in larval swimming in sea urchin.

    PubMed

    Katow, Hideki; Abe, Kouki; Katow, Tomoko; Zamani, Alemeh; Abe, Hirokazu

    2013-05-01

    The present study aimed to elucidate the development and γ-amino butyric acid (GABA)-ergic regulation of larval swimming in the sea urchin Hemicentrotus pulcherrimus by cloning glutamate decarboxylase (Hp-gad), GABAA receptor (Hp-gabrA) and GABAA receptor-associated protein (Hp-gabarap), and by performing immunohistochemistry. The regulation of larval swimming was increasingly dependent on the GABAergic system, which was active from the 2 days post-fertilization (d.p.f.) pluteus stage onwards. GABA-immunoreactive cells were detected as a subpopulation of secondary mesenchyme cells during gastrulation and eventually constituted the ciliary band and a subpopulation of blastocoelar cells during the pluteus stage. Hp-gad transcription was detected by RT-PCR during the period when Hp-Gad-positive cells were seen as a subpopulation of blastocoelar cells and on the apical side of the ciliary band from the 2 d.p.f. pluteus stage. Consistent with these observations, inhibition of GAD with 3-mercaptopropioninc acid inhibited GABA immunoreactivity and larval swimming dose dependently. Hp-gabrA amplimers were detected weakly in unfertilized eggs and 4 d.p.f. plutei but strongly from fertilized eggs to 2 d.p.f. plutei, and Hp-GabrA, together with GABA, was localized at the ciliary band in association with dopamine receptor D1 from the two-arm pluteus stage. Hp-gabarap transcription and protein expression were detected from the swimming blastula stage. Inhibition of the GABAA receptor by bicuculline inhibited larval swimming dose dependently. Inhibition of larval swimming by either 3-mercaptopropionic acid or bicuculline was more severe in older larvae (17 and 34 d.p.f. plutei) than in younger ones (1 d.p.f. prism larvae). PMID:23307803

  9. Megalodicopia hians in the Monterey submarine canyon: Distribution, larval development, and culture

    NASA Astrophysics Data System (ADS)

    Havenhand, Jon. N.; Matsumoto, George I.; Seidel, Ed

    2006-02-01

    The exclusively deep-sea ascidian family Octacnemidae comprises several genera in which the oral siphon has hypertrophied to form two large lips which create an "oral hood" capable of capturing motile prey. Megalodicopia hians is typical of this carnivorous family and has been reported to prey upon small epibenthic crustaceans. Distribution of M. hians in the Monterey Canyon system (36°45'N, 122°00'W) (California) was determined with remotely operated vehicles. M. hians was found sparsely to depths of at least 3800 m throughout the canyon; however, abundance was greatest within the oxygen-minimum zone (400-800 m). Eggs, sperm, and recently fertilized embryos were obtained repeatedly from adults returned to the laboratory in vivo, indicating that this species free-spawns routinely. Overall egg diameter (ovum plus chorion, plus follicle cells) was 175-190 μm—considerably smaller than previously reported for this species. Embryonic development at temperature and oxygen concentrations equivalent to the oxygen-minimum zone was 2-4 d and, embryos gave rise to typical phlebobranch "simple" tadpole larvae. Larval period was extremely variable, and settlement/metamorphosis occurred up to 3 months post-hatching. These results are discussed within the context of settlement-site selection and fertilization ecology of the species.

  10. Larval development of Angiostrongylus vasorum in the land snail Helix aspersa.

    PubMed

    Di Cesare, Angela; Crisi, Paolo Emidio; Bartolini, Roberto; Iorio, Raffaella; Talone, Tonino; Filippi, Laura; Traversa, Donato

    2015-10-01

    The metastrongyloid nematode Angiostrongylus vasorum affects the heart and pulmonary arteries of dogs and wild animals. Over the recent years, dog angiostrongylosis has gained great attention in the veterinary community for the expansion of its geographic range and for a rise in the number of clinical cases. Global warming, changes in phenology of mollusc intermediate hosts and movements of wild reservoirs have been evocated in the spreading of mollusc-borne parasites, including A. vasorum. The land snail Helix aspersa, a vector of other respiratory metastrongyloids, is endemic in most regions of the World, where it is a pest outside its native Mediterranean range. In the present study, the susceptibility and suitability of H. aspersa as an intermediate host of A. vasorum were investigated along with the characteristics of larval recovery and development following two different ways of inoculation, i.e. experimental (group A) vs natural infection (group B). After infections, the snails were kept at environmental conditions for 2 months. Five snails from groups A and B were randomly selected, digested and examined at 15-day intervals for 2 months. L1s, L2s and L3s were microscopically identified based on key morphological and morphometric characteristics and their identity was genetically confirmed. The results showed that A. vasorum may reach the infective stage in H. aspersa and that uptake of larvae and parasitic burden within the snails depend on the grazing capability of the molluscs. Biological and epidemiological implications are discussed. PMID:26122991

  11. Inner Ear Otolith Growth in larval Fish after Development at simulated Microgravity

    NASA Astrophysics Data System (ADS)

    Baur, U.; Hilbig, R.; Anken, R.

    It has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish via a down-regulation of carbonic anhydrase reactivity as an adaptation towards altered environmental gravity We were thus prompted to elucidate whether clinorotation would possibly yield opposite effects Therefore larval siblings of cichlid fish Oreochromis mossambicus were housed in a submersed two-dimensional clinostat Two tubes with different diameters were used 10 5 mm large tube LT and 3 5 mm small tube ST experimental time-span 10 and 7 days respectively After the experiments otoliths were dissected and their size area grown during the experiments was determined planimetrically In case of the LT-clinorotated fish both utricular and saccular otoliths lapilli and sagittae respectively were significantly smaller than those of the 1g-controls In contrast ST-maintenance resulted in significantly larger otoliths lapilli only no statistical significant difference regarding sagittae observed The results from LT-clinorotation therefore indicate that the animals had in fact received hypergravity whereas the ST-data are to be interpreted as being effected by simulated microgravity conditions In conclusion otolith growth is affected by the gravitational vector in a dose-dependent manner Acknowledgement This work was financially supported by the German Aerospace Center DLR FKZ 50 WB 9997

  12. Bt Maize Seed Mixtures for Helicoverpa zea (Lepidoptera: Noctuidae): Larval Movement, Development, and Survival on Non-transgenic Maize.

    PubMed

    Burkness, Eric C; Cira, T M; Moser, S E; Hutchison, W D

    2015-12-01

    In 2012 and 2013, field trials were conducted near Rosemount, MN, to assess the movement and development of Helicoverpa zea (Boddie) larvae on non-Bt refuge corn plants within a seed mixture of non-Bt and Bt corn. The Bt corn hybrid expressed three Bt toxins-Cry1Ab, Cry1F, and Vip3A. As the use of seed mixtures for insect resistance management (IRM) continues to be implemented, it is necessary to further characterize how this IRM approach impacts resistance development in ear-feeding Lepidopteran pests. The potential for Bt pollen movement and cross pollination of the non-Bt ears in a seed mixture may lead to Bt toxin exposure to larvae developing on those refuge ears. Larval movement and development by H. zea, feeding on non-Bt refuge plants adjacent to either transgenic Bt or non-Bt plants, were measured to investigate the potential for unintended Bt exposure. Non-Bt plants were infested with H. zea eggs and subplots were destructively sampled twice per week within each treatment to assess larval development, location, and kernel injury. Results indicate that H. zea larval movement between plants is relatively low, ranging from 2-16% of larvae, and occurs mainly after reaching the second instar. Refuge plants in seed mixtures did not produce equivalent numbers of H. zea larvae, kernel injury, and larval development differed as compared with a pure stand of non-Bt plants. This suggests that there may be costs to larvae developing on refuge plants within seed mixtures and additional studies are warranted to define potential impacts. PMID:26318006

  13. 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

  14. Ocean acidification boosts larval fish development but reduces the window of opportunity for successful settlement.

    PubMed

    Rossi, Tullio; Nagelkerken, Ivan; Simpson, Stephen D; Pistevos, Jennifer C A; Watson, Sue-Ann; Merillet, Laurene; Fraser, Peter; Munday, Philip L; Connell, Sean D

    2015-12-22

    Locating appropriate settlement habitat is a crucial step in the life cycle of most benthic marine animals. In marine fish, this step involves the use of multiple senses, including audition, olfaction and vision. To date, most investigations of larval fish audition focus on the hearing thresholds to various frequencies of sounds without testing an ecological response to such sounds. Identifying responses to biologically relevant sounds at the development stage in which orientation is most relevant is fundamental. We tested for the existence of ontogenetic windows of reception to sounds that could act as orientation cues with a focus on vulnerability to alteration by human impacts. Here we show that larvae of a catadromous fish species (barramundi, Lates calcarifer) were attracted towards sounds from settlement habitat during a surprisingly short ontogenetic window of approximately 3 days. Yet, this auditory preference was reversed in larvae reared under end-of-century levels of elevated CO2, such that larvae are repelled from cues of settlement habitat. These future conditions also reduced the swimming speeds and heightened the anxiety levels of barramundi. Unexpectedly, an acceleration of development and onset of metamorphosis caused by elevated CO2 were not accompanied by the earlier onset of attraction towards habitat sounds. This mismatch between ontogenetic development and the timing of orientation behaviour may reduce the ability of larvae to locate habitat or lead to settlement in unsuitable habitats. The misinterpretation of key orientation cues can have implications for population replenishment, which are only exacerbated when ontogenetic development decouples from the specific behaviours required for location of settlement habitats. PMID:26674946

  15. Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development

    PubMed Central

    Walker, Benjamin S.; Lassiter, Christopher S.; Jónsson, Zophonías O.

    2016-01-01

    The development of craniofacial skeletal structures requires well-orchestrated tissue interactions controlled by distinct molecular signals. Disruptions in normal function of these molecular signals have been associated with a wide range of craniofacial malformations. A pathway mediated by estrogens is one of those molecular signals that plays role in formation of bone and cartilage including craniofacial skeletogenesis. Studies in zebrafish have shown that while higher concentrations of 17-β estradiol (E2) cause severe craniofacial defects, treatment with lower concentrations result in subtle changes in head morphology characterized with shorter snouts and flatter faces. The molecular basis for these morphological changes, particularly the subtle skeletal effects mediated by lower E2 concentrations, remains unexplored. In the present study we address these effects at a molecular level by quantitative expression analysis of sets of candidate genes in developing heads of zebrafish larvae treated with two different E2 concentrations. To this end, we first validated three suitable reference genes, ppia2, rpl8 and tbp, to permit sensitive quantitative real-time PCR analysis. Next, we profiled the expression of 28 skeletogenesis-associated genes that potentially respond to estrogen signals and play role in craniofacial development. We found E2 mediated differential expression of genes involved in extracellular matrix (ECM) remodelling, mmp2/9/13, sparc and timp2a, as well as components of skeletogenic pathways, bmp2a, erf, ptch1/2, rankl, rarab and sfrp1a. Furthermore, we identified a co-expressed network of genes, including cpn1, dnajc3, esr1, lman1, rrbp1a, ssr1 and tram1 with a stronger inductive response to a lower dose of E2 during larval head development. PMID:27069811

  16. Investigation of the effects of estrogen on skeletal gene expression during zebrafish larval head development.

    PubMed

    Pashay Ahi, Ehsan; Walker, Benjamin S; Lassiter, Christopher S; Jónsson, Zophonías O

    2016-01-01

    The development of craniofacial skeletal structures requires well-orchestrated tissue interactions controlled by distinct molecular signals. Disruptions in normal function of these molecular signals have been associated with a wide range of craniofacial malformations. A pathway mediated by estrogens is one of those molecular signals that plays role in formation of bone and cartilage including craniofacial skeletogenesis. Studies in zebrafish have shown that while higher concentrations of 17-β estradiol (E 2) cause severe craniofacial defects, treatment with lower concentrations result in subtle changes in head morphology characterized with shorter snouts and flatter faces. The molecular basis for these morphological changes, particularly the subtle skeletal effects mediated by lower E 2 concentrations, remains unexplored. In the present study we address these effects at a molecular level by quantitative expression analysis of sets of candidate genes in developing heads of zebrafish larvae treated with two different E 2 concentrations. To this end, we first validated three suitable reference genes, ppia2, rpl8 and tbp, to permit sensitive quantitative real-time PCR analysis. Next, we profiled the expression of 28 skeletogenesis-associated genes that potentially respond to estrogen signals and play role in craniofacial development. We found E 2 mediated differential expression of genes involved in extracellular matrix (ECM) remodelling, mmp2/9/13, sparc and timp2a, as well as components of skeletogenic pathways, bmp2a, erf, ptch1/2, rankl, rarab and sfrp1a. Furthermore, we identified a co-expressed network of genes, including cpn1, dnajc3, esr1, lman1, rrbp1a, ssr1 and tram1 with a stronger inductive response to a lower dose of E 2 during larval head development. PMID:27069811

  17. Drosophila alcohol dehydrogenase: developmental studies on cryptic variant lines.

    PubMed

    Miglani, G S; Ampy, F R

    1981-10-01

    Thirty-five cryptic variant lines were used to examine the mechanisms involved in genetic modulation of alcohol metabolism in Drosophila. Late third-instar larval, preemergence pupal, and adult stages cultured at 18 and 28 C were examined. Spectrophotometric analyses for native alcohol dehydrogenase (ADH) activity and residual ADH activity after treatment with guanidine hydrochloride and heat were performed. Differential response of cryptic variants to treatment with the denaturants during development suggested that this variation may have an adaptive significance. PMID:6800354

  18. Coordinated Development of Muscles and Tendon-Like Structures: Early Interactions in the Drosophila Leg.

    PubMed

    Soler, Cedric; Laddada, Lilia; Jagla, Krzysztof

    2016-01-01

    The formation of the musculoskeletal system is a remarkable example of tissue assembly. In both vertebrates and invertebrates, precise connectivity between muscles and skeleton (or exoskeleton) via tendons or equivalent structures is fundamental for movement and stability of the body. The molecular and cellular processes underpinning muscle formation are well-established and significant advances have been made in understanding tendon development. However, the mechanisms contributing to proper connection between these two tissues have received less attention. Observations of coordinated development of tendons and muscles suggest these tissues may interact during the different steps in their development. There is growing evidence that, depending on animal model and muscle type, these interactions can take place from progenitor induction to the final step of the formation of the musculoskeletal system. Here, we briefly review and compare the mechanisms behind muscle and tendon interaction throughout the development of vertebrates and Drosophila before going on to discuss our recent findings on the coordinated development of muscles and tendon-like structures in Drosophila leg. By altering apodeme formation (the functional Drosophila equivalent of tendons in vertebrates) during the early steps of leg development, we affect the spatial localization of subsequent myoblasts. These findings provide the first evidence of the developmental impact of early interactions between muscle and tendon-like precursors, and confirm the appendicular Drosophila muscle system as a valuable model for studying these processes. PMID:26869938

  19. Coordinated Development of Muscles and Tendon-Like Structures: Early Interactions in the Drosophila Leg

    PubMed Central

    Soler, Cedric; Laddada, Lilia; Jagla, Krzysztof

    2016-01-01

    The formation of the musculoskeletal system is a remarkable example of tissue assembly. In both vertebrates and invertebrates, precise connectivity between muscles and skeleton (or exoskeleton) via tendons or equivalent structures is fundamental for movement and stability of the body. The molecular and cellular processes underpinning muscle formation are well-established and significant advances have been made in understanding tendon development. However, the mechanisms contributing to proper connection between these two tissues have received less attention. Observations of coordinated development of tendons and muscles suggest these tissues may interact during the different steps in their development. There is growing evidence that, depending on animal model and muscle type, these interactions can take place from progenitor induction to the final step of the formation of the musculoskeletal system. Here, we briefly review and compare the mechanisms behind muscle and tendon interaction throughout the development of vertebrates and Drosophila before going on to discuss our recent findings on the coordinated development of muscles and tendon-like structures in Drosophila leg. By altering apodeme formation (the functional Drosophila equivalent of tendons in vertebrates) during the early steps of leg development, we affect the spatial localization of subsequent myoblasts. These findings provide the first evidence of the developmental impact of early interactions between muscle and tendon-like precursors, and confirm the appendicular Drosophila muscle system as a valuable model for studying these processes. PMID:26869938

  20. The glial investment of the adult and developing antennal lobe of Drosophila

    PubMed Central

    Oland, Lynne A.; Biebelhausen, John P.; Tolbert, Leslie P.

    2009-01-01

    In recent years, the Drosophila olfactory system, with its unparalleled opportunities for genetic dissection of development and functional organization, has been used to study the development of central olfactory neurons and the molecular basis of olfactory coding. The results of these studies have been interpreted in the absence of a detailed understanding of the steps in maturation of glial cells in the antennal lobe. Here, we present a high-resolution study of the glia associated with olfactory glomeruli in adult and developing antennal lobes. The study provides a basis for comparison of findings in Drosophila with those in the moth Manduca sexta that indicate a critical role for glia in antennal lobe development. Using flies expressing GFP under a Nervana2 driver to visualize glia for confocal microscopy, and probing at higher resolution with the electron microscope, we find that glial development in Drosophila differs markedly from that in moths: glial cell bodies remain in a rind around the glomerular neuropil; glial processes ensheathe axon bundles in the nerve layer but likely contribute little to axonal sorting; their processes insinuate between glomeruli only very late and then form only a sparse, open network around each glomerulus; and glial processes invade the synaptic neuropil. Taking our results in the context of previous studies, we conclude that glial cells in the developing Drosophila antennal lobe are unlikely to play a strong role in either axonal sorting or glomerulus stabilization and that in the adult, glial processes do not electrically isolate glomeruli from their neighbors. PMID:18537134

  1. Ethanol Affects the Development of Sensory Hair Cells in Larval Zebrafish (Danio rerio)

    PubMed Central

    Matsui, Jonathan I.

    2013-01-01

    Children born to mothers with substantial alcohol consumption during pregnancy can present a number of morphological, cognitive, and sensory abnormalities, including hearing deficits, collectively known as fetal alcohol syndrome (FAS). The goal of this study was to determine if the zebrafish lateral line could be used to study sensory hair cell abnormalities caused by exposure to ethanol during embryogenesis. Some lateral line sensory hair cells are present at 2 days post-fertilization (dpf) and are functional by 5 dpf. Zebrafish embryos were raised in fish water supplemented with varying concentrations of ethanol (0.75%–1.75% by volume) from 2 dpf through 5 dpf. Ethanol treatment during development resulted in many physical abnormalities characteristic of FAS in humans. Also, the number of sensory hair cells decreased as the concentration of ethanol increased in a dose-dependent manner. The dye FM 1-43FX was used to detect the presence of functional mechanotransduction channels. The percentage of FM 1-43-labeled hair cells decreased as the concentration of ethanol increased. Methanol treatment did not affect the development of hair cells. The cell cycle markers proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU) demonstrated that ethanol reduced the number of sensory hair cells, as a consequence of decreased cellular proliferation. There was also a significant increase in the rate of apoptosis, as determined by TUNEL-labeling, in neuromasts following ethanol treatment during larval development. Therefore, zebrafish are a useful animal model to study the effects of hair cell developmental disorders associated with FAS. PMID:24324841

  2. Proteome analysis of hemolymph changes during the larval to pupal development stages of honeybee workers (Apis mellifera ligustica).

    PubMed

    Woltedji, Dereje; Fang, Yu; Han, Bin; Feng, Mao; Li, Rongli; Lu, Xiaoshan; Li, Jianke

    2013-11-01

    Hemolymph is vital for the flow and transportation of nutrients, ions, and hormones in the honey bee and plays role in innate immune defense. The proteome of the hemolymph changes over the life of a honey bee, but many of these changes are not well characterized, including changes during the life cycle transition from the larval to pupal stages of workers. We used two-dimensional gel electrophoresis, mass spectrometry, bioinformatics, and Western blot to analyze the proteome changes of the honeybee hemolymph during the transition from newly hatched larvae to five-day-old pupae. Of the 49 nonredundant proteins that changed in abundance (identified by 80 protein spots), 29 (59.2%) and 20 (40.8%) were strongly expressed in the larvae and the pupae, respectively. The larval hemolymph had high expressions of major royal jelly proteins and proteins related to metabolism of carbohydrates and energy, folding activities, development, and the cytoskeleton and antioxidant systems. Proteins involved in food storage and the metabolism of fatty acids and amino acids were abundantly expressed during the late larval to pupal development stages. The proteins expressed by the young larvae are used to enhance their development process and as a temporal innate immune protection mechanism until they gain immunity with age development. The pupae use more energy storage related proteins as they prepare for their non-diet-driven pupation. Our data provide new evidence that changes in the hemolymph at the proteome level match the processes during life transitions in the honeybee. PMID:24006915

  3. Reproduction and development in Halocaridina rubra Holthuis, 1963 (Crustacea: Atyidae) clarifies larval ecology in the Hawaiian anchialine ecosystem.

    PubMed

    Havird, Justin C; Vaught, Rebecca C; Weese, David A; Santos, Scott R

    2015-10-01

    Larvae in aquatic habitats often develop in environments different from those they inhabit as adults. Shrimp in the Atyidae exemplify this trend, as larvae of many species require salt or brackish water for development, while adults are freshwater-adapted. An exception within the Atyidae family is the "anchialine clade," which are euryhaline as adults and endemic to habitats with subterranean fresh and marine water influences. Although the Hawaiian anchialine atyid Halocaridina rubra is a strong osmoregulator, its larvae have never been observed in nature. Moreover, larval development in anchialine species is poorly studied. Here, reproductive trends in laboratory colonies over a 5-y period are presented from seven genetic lineages and one mixed population of H. rubra; larval survivorship under varying salinities is also discussed. The presence and number of larvae differed significantly among lineages, with the mixed population being the most prolific. Statistical differences in reproduction attributable to seasonality also were identified. Larval survivorship was lowest (12% settlement rate) at a salinity approaching fresh water and significantly higher in brackish and seawater (88% and 72%, respectively). Correlated with this finding, identifiable gills capable of ion transport did not develop until metamorphosis into juveniles. Thus, early life stages of H. rubra are apparently excluded from surface waters, which are characterized by lower and fluctuating salinities. Instead, these stages are restricted to the subterranean (where there is higher and more stable salinity) portion of Hawaii's anchialine habitats due to their inability to tolerate low salinities. Taken together, these data contribute to the understudied area of larval ecology in the anchialine ecosystem. PMID:26504154

  4. Drosophila Avoids Parasitoids by Sensing Their Semiochemicals via a Dedicated Olfactory Circuit

    PubMed Central

    Ebrahim, Shimaa A. M.; Dweck, Hany K. M.; Stökl, Johannes; Hofferberth, John E.; Trona, Federica; Weniger, Kerstin; Rybak, Jürgen; Seki, Yoichi; Stensmyr, Marcus C.; Sachse, Silke; Hansson, Bill S.; Knaden, Markus

    2015-01-01

    Detecting danger is one of the foremost tasks for a neural system. Larval parasitoids constitute clear danger to Drosophila, as up to 80% of fly larvae become parasitized in nature. We show that Drosophila melanogaster larvae and adults avoid sites smelling of the main parasitoid enemies, Leptopilina wasps. This avoidance is mediated via a highly specific olfactory sensory neuron (OSN) type. While the larval OSN expresses the olfactory receptor Or49a and is tuned to the Leptopilina odor iridomyrmecin, the adult expresses both Or49a and Or85f and in addition detects the wasp odors actinidine and nepetalactol. The information is transferred via projection neurons to a specific part of the lateral horn known to be involved in mediating avoidance. Drosophila has thus developed a dedicated circuit to detect a life-threatening enemy based on the smell of its semiochemicals. Such an enemy-detecting olfactory circuit has earlier only been characterized in mice and nematodes. PMID:26674493

  5. 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.

  6. Yolk-sac larval development of the substrate-brooding cichlid Archocentrus nigrofasciatus in relation to temperature.

    PubMed

    Vlahos, Nikolaos; Vasilopoulos, Michael; Mente, Eleni; Hotos, George; Katselis, George; Vidalis, Kosmas

    2015-09-01

    In order to conserve and culture the cichlid fish Archocentrus nigrofasciatus, more information about its reproductive biology and its larval behavior and morphogenesis is necessary. Currently, temperatures ranging from 21 to 27 °C are used in ornamental aquaculture hatcheries. Lower temperatures are preferred to reduce the costs of water heating, and 23 °C is usually the selected temperature. However, there is limited information on culturing protocols for ornamental species and most of the information generated on this topic remains scarce. Thus, the present study examines the morphological development of Archocentrus nigrofasciatus during the yolk-sac period up to the age of 100 h post-hatching in relation to 2 temperature regimes used in ornamental aquaculture: a temperature of 27 °C (thermal optimum) and a decreased temperature of 23 °C (thermal tolerance). The results of this study suggest that the 27 °C temperature generates intense morphological changes in yolk-sac development in a shorter period. This has advantages as it reduces the time of yolk-sac larval development, and, thus, minimizes the transition phase to exogenous feeding and maximizes the efficiency at which yolk is converted into body tissues. The present paper provides necessary information to produce freshwater ornamental fish with better practices so as to increase larval survival and capitalize on time for growth. PMID:26201370

  7. Inbreeding Effects in Families of Ostrinia nubilalis (Lepidoptera: Crambidae): Larval Development in Laboratory Bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inbreeding depression of laboratory-reared insects has the potential to affect their larval performance and reproductive output. Two studies of laboratory-reared colonies of Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae) were conducted to assess whether inbreeding affected a laboratory bioass...

  8. Efficacy and longevity of the newly developed catnip oil microcapsules against stable fly oviposition larval growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The stable fly, Stomoxys calcitrans (Diptera: Muscidae), is one of the most important pests of cattle and costs U.S. cattle producers billions of dollars in losses annually. In this study, the efficacy of catnip oil encapsulated in gelatin in oviposition deterrence and larval growth inhibition in st...

  9. Efficacy and longevity of newly developed catnip oil microcapsules against stable fly oviposition and larval growth.

    PubMed

    Zhu, J J; Wienhold, B J; Wehrle, J; Davis, D; Chen, H; Taylor, D; Friesen, K; Zurek, L

    2014-06-01

    The stable fly, Stomoxys calcitrans (Diptera: Muscidae), is one of the most important pests of cattle and costs U.S. cattle producers billions of dollars in losses annually. In this study, the efficacy of catnip oil encapsulated in gelatin in oviposition deterrence and larval growth inhibition in stable flies was examined under laboratory conditions. More than 98% inhibition of stable fly larval growth and female oviposition was observed in larval and oviposition media treated with encapsulated catnip oil (0.5 g). Further, dose-response tests showed that as little as 0.1 g of encapsulated catnip oil provided > 85% oviposition deterrence. The release of nepetalactones from the capsules was more rapid when the capsules were placed on a moist substrate rather than a dry substrate. Encapsulated catnip oil also exhibited antibacterial activity, supporting the hypothesis that its inhibition of larval growth may be based on its killing of the bacteria on which larvae feed. The use of encapsulated catnip oil can provide an alternative control strategy for stable fly management. PMID:24111783

  10. Dispersal of stable flies (Diptera: Muscidae) from larval development sites in a Nebraska landscape

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seven mark recapture studies of stable flies wer conducted in a mixed agricultural environment in northeastern Nebraska. Larval developmental sites were marked by dusting the surface with fluorescent pigments and stable flies were recaptured with grids of Alsynite sticky traps. Recaptured flies were...

  11. Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection.

    PubMed

    Mitchell, Mairi C; Tzelos, Thomas; Handel, Ian; McWilliam, Hamish E G; Hodgkinson, Jane E; Nisbet, Alasdair J; Kharchenko, Vitaliy O; Burgess, Stewart T G; Matthews, Jacqueline B

    2016-07-01

    Cyathostomins are ubiquitous nematodes of horses. Once ingested, they can spend a substantial time as encysted larvae in the intestinal wall. The larvae can comprise up to 90% of the total burden, with up to several million worms reported in individuals. These stages can emerge in large numbers to cause life-threatening colitis. Direct methods for detection of encysted larval burdens in live horses do not exist. Previously, two antigen complexes were identified as promising markers for infection. A component of these, cyathostomin gut associated larval antigen-1 (Cy-GALA-1), was identified following immunoscreening of a complementary DNA library. Serum immunoglobulin G(T) (IgG(T)) responses to Cy-GALA-1 were shown to inform on larval infection. Sequence analysis of polymerase chain reaction products amplified from individual worms indicated that Cy-GALA-1 was derived from Cyathostomum pateratum. As cyathostomin infections always comprise multiple species, a diagnostic test must account for this. Here, segments of the Cy-gala gene were isolated from four common species, Cyathostomum catinatum, Cylicocyclus ashworthi, Cylicostephanus goldi and Cylicostephanus longibursatus, and the associated proteins expressed in recombinant form. The specificity and immunogenicity of each protein was confirmed. Each protein was assessed by enzyme linked immuno sorbent assay (ELISA) for its ability for informing on the presence of encysted larval infection and the level of burden. PMID:27174468

  12. Control of larval and egg development in Aedes aegypti with RNA interference against juvenile hormone acid methyl transferase.

    PubMed

    Van Ekert, Evelien; Powell, Charles A; Shatters, Robert G; Borovsky, Dov

    2014-11-01

    RNA interference (RNAi) is a powerful approach for elucidating gene functions in a variety of organisms, including mosquitoes and many other insects. Little has been done, however, to harness this approach in order to control adult and larval mosquitoes. Juvenile hormone (JH) plays a pivotal role in the control of reproduction in adults and metamorphism in larval mosquitoes. This report describes an approach to control Aedes aegypti using RNAi against JH acid methyl transferase (AeaJHAMT), the ultimate enzyme in the biosynthetic pathway of JH III that converts JH acid III (JHA III) into JH III. In female A. aegypti that were injected or fed jmtA dsRNA targeting the AeaJHAMT gene (jmtA) transcript, egg development was inhibited in 50% of the treated females. In mosquito larvae that were fed transgenic Pichia pastoris cells expressing long hair pin (LHP) RNA, adult eclosion was delayed by 3 weeks causing high mortality. Northern blot analyses and qPCR studies show that jmtA dsRNA causes inhibition of jmtA transcript in adults and larvae, which is consistent with the observed inhibition of egg maturation and larval development. Taken together, these results suggest that jmtA LHP RNA expressed in heat inactivated genetically modified P. pastoris cells could be used to control mosquito populations in the marsh. PMID:25111689

  13. Effect of CO2-related acidification on aspects of the larval development of the European lobster, Homarus gammarus (L.)

    NASA Astrophysics Data System (ADS)

    Arnold, K. E.; Findlay, H. S.; Spicer, J. I.; Daniels, C. L.; Boothroyd, D.

    2009-03-01

    Oceanic uptake of anthropogenic CO2 results in a reduction in pH termed "Ocean Acidification" (OA). Comparatively little attention has been given to the effect of OA on the early life history stages of marine animals. Consequently, we investigated the effect of culture in CO2-acidified sea water (approx. 1200 ppm, i.e. average values predicted using IPCC 2007 A1F1 emissions scenarios for year 2100) on early larval stages of an economically important crustacean, the European lobster Homarus gammarus. Culture in CO2-acidified sea water did not significantly affect carapace length or development of H. gammarus. However, there was a reduction in carapace mass during the final stage of larval development in CO2-acidified sea water. This co-occurred with a reduction in exoskeletal mineral (calcium and magnesium) content of the carapace. As the control and high CO2 treatments were not undersaturated with respect to any of the calcium carbonate polymorphs measured, the physiological alterations we record are most likely the result of acidosis or hypercapnia interfering with normal homeostatic function, and not a direct impact on the carbonate supply-side of calcification per se. Thus despite there being no observed effect on survival, carapace length, or zoeal progression, OA related (indirect) disruption of calcification and carapace mass might still adversely affect the competitive fitness and recruitment success of larval lobsters with serious consequences for population dynamics and marine ecosystem function.

  14. The Implications of Temperature-Mediated Plasticity in Larval Instar Number for Development within a Marine Invertebrate, the Shrimp Palaemonetes varians

    PubMed Central

    Oliphant, Andrew; Hauton, Chris; Thatje, Sven

    2013-01-01

    Variations in larval instar number are common among arthropods. Here, we assess the implications of temperature-mediated variations in larval instar number for larval development time, larval growth rates, and juvenile dry weight within the palaemonid shrimp, Palaemonetes varians. In contrast with previous literature, which focuses on terrestrial arthropods, particularly model and pest species often of laboratory lines, we use wild shrimp, which differ in their life history from previous models. Newly-hatched P. varians larvae were first reared at 5, 10, 17, 25, and 30°C to assess their thermal scope for development. Larvae developed at 17, 25, and 30°C. At higher temperatures, larvae developed through fewer larval instars. Two dominant developmental pathways were observed; a short pathway of four instars and a long pathway of five instars. Longer developmental pathways of six to seven instars were rarely observed (mostly at lower temperatures) and consisted of additional instars as ‘repeat’ instars; i.e. little developmental advance over the preceding instar. To assess the implications of temperature-mediated variation in larval instar number, newly-hatched larvae were then reared at 15, 20, and 25°C. Again, the proportion of larvae developing through four instars increased with temperature. At all temperatures, larval development time and juvenile dry weight were greater for larvae developing through five instars. Importantly, because of the increasing proportion of larvae developing through four instars with increasing temperature, larval traits associated with this pathway (reduced development time and juvenile dry weight) became more dominant. As a consequence of increasing growth rate with temperature, and the shift in the proportion of larvae developing through four instars, juvenile dry weight was greatest at intermediate temperatures (20°C). We conclude that at settlement P. varians juveniles do not follow the temperature-size rule; this is of

  15. Regional immune responses with stage-specific antigen recognition profiles develop in lymph nodes of pigs following Ascaris suum larval migration.

    PubMed

    Jungersen, G; Eriksen, L; Nansen, P; Lind, P; Rasmussen, T; Meeusen, E N

    2001-04-01

    The early life-cycle of the pig round worm, Ascaris suum, involves well-defined larval development in the liver, lungs and finally the small intestine. Distinct regional immune responses to larval antigens of A. suum were observed in the draining lymph nodes of immunized and challenged pigs during larval migration. This was reflected in a transient enlargement of the stimulated lymph nodes, due to increases in numbers of B cells and CD4 T cells, and the production of A. suum-specific antibody by antibody secreting cell (ASC) cultures. Larval antigen recognition pattern of antibodies in serum, bile and draining lymph node ASC culture supernatant (ASC-probes) was examined by immunoblotting. This revealed distinct organ-specific recognition patterns of larval-specific antigens by the draining lymph nodes at different times after challenge. In particular, an early larval 42 kDa antigen was recognized specifically by ASC-probes of the liver lymph nodes at 7 but not 14 days postchallenge (pc) which was not detected in other lymph nodes, serum or bile of the same pig. Similarly, a late larval antigen of 34 kDa was uniquely detected by lung and jejunal ASC-probes at 14 days pc. These observations demonstrate how development of distinct regional immune responses in tissues with different antigen stimulation can be monitored with ASC-probes and flow cytometry. PMID:11298295

  16. Phosphoproteome analysis during larval development and metamorphosis in the spionid polychaete Pseudopolydora vexillosa

    PubMed Central

    2011-01-01

    Background The metamorphosis of the spionid polychaete Pseudopolydora vexillosa includes spontaneous settlement onto soft-bottom habitats and morphogenesis that can be completed in a very short time. A previous study on the total changes to the proteome during the various developmental stages of P. vexillosa suggested that little or no de novo protein synthesis occurs during metamorphosis. In this study, we used multicolor fluorescence detection of proteins in 2-D gels for differential analysis of proteins and phosphoproteins to reveal the dynamics of post-translational modification proteins in this species. A combination of affinity chromatography, 2D-PAGE, and mass spectrometry was used to identify the phosphoproteins in pre-competent larvae, competent larvae, and newly metamorphosed juveniles. Results We reproducibly detected 210, 492, and 172 phosphoproteins in pre-competent larvae, competent larvae, and newly metamorphosed juveniles, respectively. The highest percentage of phosphorylation was observed during the competent larval stage. About 64 stage-specific phosphoprotein spots were detected in the competent stage, and 32 phosphoproteins were found to be significantly differentially expressed in the three stages. We identified 38 phosphoproteins, 10 of which were differentially expressed during metamorphosis. These phosphoproteins belonged to six categories of biological processes: (1) development, (2) cell differentiation and integrity, (3) transcription and translation, (4) metabolism, (5) protein-protein interaction and proteolysis, and (6) receptors and enzymes. Conclusion This is the first study to report changes in phosphoprotein expression patterns during the metamorphosis of the marine polychaete P. vexillosa. The higher degree of phosphorylation during the process of attaining competence to settle and metamorphose may be due to fast morphological transitions regulated by various mechanisms. Our data are consistent with previous studies showing a

  17. Genetic analysis of contributions of dorsal group and JAK-Stat92E pathway genes to larval hemocyte concentration and the egg encapsulation response in Drosophila.

    PubMed Central

    Sorrentino, Richard Paul; Melk, Jonathan P; Govind, Shubha

    2004-01-01

    Drosophila larvae defend themselves against parasitoid wasps by completely surrounding the egg with layers of specialized hemocytes called lamellocytes. Similar capsules of lamellocytes, called melanotic capsules, are also formed around "self" tissues in larvae carrying gain-of-function mutations in Toll and hopscotch. Constitutive differentiation of lamellocytes in larvae carrying these mutations is accompanied by high concentrations of plasmatocytes, the major hemocyte class in uninfected control larvae. The relative contributions of hemocyte concentration vs. lamellocyte differentiation to wasp egg encapsulation are not known. To address this question, we used Leptopilina boulardi to infect more than a dozen strains of host larvae harboring a wide range of hemocyte densities. We report a significant correlation between hemocyte concentration and encapsulation capacity among wild-type larvae and larvae heterozygous for mutations in the Hopscotch-Stat92E and Toll-Dorsal pathways. Larvae carrying loss-of-function mutations in Hopscotch, Stat92E, or dorsal group genes exhibit significant reduction in encapsulation capacity. Larvae carrying loss-of-function mutations in dorsal group genes (including Toll and tube) have reduced hemocyte concentrations, whereas larvae deficient in Hopscotch-Stat92E signaling do not. Surprisingly, unlike hopscotch mutants, Toll and tube mutants are not compromised in their ability to generate lamellocytes. Our results suggest that circulating hemocyte concentration and lamellocyte differentiation constitute two distinct physiological requirements of wasp egg encapsulation and Toll and Hopscotch proteins serve distinct roles in this process. PMID:15082553

  18. Modeling congenital disease and inborn errors of development in Drosophila melanogaster

    PubMed Central

    Moulton, Matthew J.; Letsou, Anthea

    2016-01-01

    ABSTRACT Fly models that faithfully recapitulate various aspects of human disease and human health-related biology are being used for research into disease diagnosis and prevention. Established and new genetic strategies in Drosophila have yielded numerous substantial successes in modeling congenital disorders or inborn errors of human development, as well as neurodegenerative disease and cancer. Moreover, although our ability to generate sequence datasets continues to outpace our ability to analyze these datasets, the development of high-throughput analysis platforms in Drosophila has provided access through the bottleneck in the identification of disease gene candidates. In this Review, we describe both the traditional and newer methods that are facilitating the incorporation of Drosophila into the human disease discovery process, with a focus on the models that have enhanced our understanding of human developmental disorders and congenital disease. Enviable features of the Drosophila experimental system, which make it particularly useful in facilitating the much anticipated move from genotype to phenotype (understanding and predicting phenotypes directly from the primary DNA sequence), include its genetic tractability, the low cost for high-throughput discovery, and a genome and underlying biology that are highly evolutionarily conserved. In embracing the fly in the human disease-gene discovery process, we can expect to speed up and reduce the cost of this process, allowing experimental scales that are not feasible and/or would be too costly in higher eukaryotes. PMID:26935104

  19. Modeling congenital disease and inborn errors of development in Drosophila melanogaster.

    PubMed

    Moulton, Matthew J; Letsou, Anthea

    2016-03-01

    Fly models that faithfully recapitulate various aspects of human disease and human health-related biology are being used for research into disease diagnosis and prevention. Established and new genetic strategies in Drosophila have yielded numerous substantial successes in modeling congenital disorders or inborn errors of human development, as well as neurodegenerative disease and cancer. Moreover, although our ability to generate sequence datasets continues to outpace our ability to analyze these datasets, the development of high-throughput analysis platforms in Drosophila has provided access through the bottleneck in the identification of disease gene candidates. In this Review, we describe both the traditional and newer methods that are facilitating the incorporation of Drosophila into the human disease discovery process, with a focus on the models that have enhanced our understanding of human developmental disorders and congenital disease. Enviable features of the Drosophila experimental system, which make it particularly useful in facilitating the much anticipated move from genotype to phenotype (understanding and predicting phenotypes directly from the primary DNA sequence), include its genetic tractability, the low cost for high-throughput discovery, and a genome and underlying biology that are highly evolutionarily conserved. In embracing the fly in the human disease-gene discovery process, we can expect to speed up and reduce the cost of this process, allowing experimental scales that are not feasible and/or would be too costly in higher eukaryotes. PMID:26935104

  20. Interactions of temperature and steroids on larval growth, development, and metamorphosis in a toad (Bufo boreas).

    PubMed

    Hayes, T; Chan, R; Licht, P

    1993-07-01

    The effects of temperature and steroids [testosterone (T), estradiol (E2), and corticosterone (B)] on premetamorphic growth and development were investigated in the toad (Bufo boreas). The effects of steroids were both temperature and age dependent. In the first experiment, steroids (1.1-1.4 microM) were administered by dissolving them in the water beginning 1 day after hatching at 22 degrees C or 27 degrees C. At 22 degrees C, B inhibited growth (P < 0.001) but had no significant effect on development. Forelegs never emerged in B-treated animals and all died before complete tail resorption. Discontinuation of B treatment allowed normal growth and metamorphosis, but the resulting post-metamorphic animals were significantly shorter (snout-vent length, P < 0.001) than after other treatments. At 22 degrees C, T and E2 had no effect on larval growth and development or size at metamorphosis (P > 0.05), but T induced early foreleg emergence (FLE) (P < 0.005). At 27 degrees C, B was fatal after 2 weeks of treatment, and T and E2 inhibited growth (P < 0.001) and development (P < 0.001), but did not affect time to FLE. In a second experiment at 27 degrees C, treatment with 1.1 microM B starting 15 days after hatching induced early metamorphic events (P < 0.001), such as tail resorption and emergence of the left foreleg (but not the right), but jaw and head restructuring failed to occur. All B-treated animals died before complete tail resorption. In a third experiment, 0.275 and 1.11 microM B, starting at day 43 (stage 43), induced early FLE (P < 0.05) and decreased snout-vent length at tail resorption (P < 0.005) without a dose effect. A higher dose of B (4.44 microM) decreased snout-vent length at tail resorption and time to FLE (P < 0.05) but did not affect body weight at metamorphosis (P > 0.05). Animals in this experiment survived to complete tail resorption and transformed normally. The actions of B in these experiments closely resemble those observed with

  1. Mosquito larval habitat mapping using remote sensing and GIS: Implications of coalbed methane development and West Nile virus

    SciTech Connect

    Zou, L.; Miller, S.N.; Schmidtmann, E.T.

    2006-09-15

    Potential larval habitats of the mosquito Culex tarsalis (Coquillett), implicated as a primary vector of West Nile virus in Wyoming, were identified using integrated remote sensing and geographic information system (GIS) analyses. The study area is in the Powder River Basin of north central Wyoming, an area that has been undergoing a significant increase in coalbed methane gas extractions since the late 1990s. Large volumes of water are discharged, impounded, and released during the extraction of methane gas, creating aquatic habitats that have the potential to support immature mosquito development. Landsat TM and ETM + data were initially classified into spectrally distinct water and vegetation classes, which were in turn used to identify suitable larval habitat sites. This initial habitat classification was refined using knowledge-based GIS techniques requiring spatial data layers for topography, streams, and soils to reduce the potential for overestimation of habitat. Accuracy assessment was carried out using field data and high-resolution aerial photography commensurate with one of the Landsat images. The classifier can identify likely habitat for ponds larger than 0.8 ha (2 acres) with generally satisfactory results (72.1%) with a lower detection limit of approximate to 0.4 ha (1 acre). Results show a 75% increase in potential larval habitats from 1999 to 2004 in the study area, primarily because of the large increase in small coalbed methane water discharge ponds. These results may facilitate mosquito abatement programs in the Powder River Basin with the potential for application throughout the state and region.

  2. Effect of CO2-related acidification on aspects of the larval development of the European lobster, Homarus gammarus (L.)

    NASA Astrophysics Data System (ADS)

    Arnold, K. E.; Findlay, H. S.; Spicer, J. I.; Daniels, C. L.; Boothroyd, D.

    2009-08-01

    Oceanic uptake of anthropogenic CO2 results in a reduction in pH termed "Ocean Acidification" (OA). Comparatively little attention has been given to the effect of OA on the early life history stages of marine animals. Consequently, we investigated the effect of culture in CO2-acidified sea water (approx. 1200 ppm, i.e. average values predicted using IPCC 2007 A1F1 emissions scenarios for year 2100) on early larval stages of an economically important crustacean, the European lobster Homarus gammarus. Culture in CO2-acidified sea water did not significantly affect carapace length of H. gammarus. However, there was a reduction in carapace mass during the final stage of larval development in CO2-acidified sea water. This co-occurred with a reduction in exoskeletal mineral (calcium and magnesium) content of the carapace. As the control and high CO2 treatments were not undersaturated with respect to any of the calcium carbonate polymorphs measured, the physiological alterations we record are most likely the result of acidosis or hypercapnia interfering with normal homeostatic function, and not a direct impact on the carbonate supply-side of calcification per se. Thus despite there being no observed effect on survival, carapace length, or zoeal progression, OA related (indirect) disruption of calcification and carapace mass might still adversely affect the competitive fitness and recruitment success of larval lobsters with serious consequences for population dynamics and marine ecosystem function.

  3. The Development of Motor Coordination in Drosophila Embryos

    PubMed Central

    Crisp, Sarah; Evers, Jan Felix; Fiala, André; Bate, Michael

    2012-01-01

    We use non-invasive muscle imaging to study onset of motor activity and emergence of coordinated movement in Drosophila embryos. Earliest movements are myogenic and neurally controlled muscle contractions first appear with the onset of bursting activity 17 hours after egg laying. Initial episodes of activity are poorly organised and coordinated crawling sequences only begin to appear after a further hour of bursting. Thus network performance improves during this first period of activity. The embryo continues to exhibit bursts of crawling like sequences until shortly before hatching, while other reflexes also mature. Bursting does not begin as a reflex response to sensory input but appears to reflect the onset of spontaneous activity in the motor network. It does not require GABA-ergic transmission, and using a light activated channel to excite the network we demonstrate activity dependent depression that may cause burst termination. PMID:18927150

  4. The Development of Germline Stem Cells in Drosophila

    PubMed Central

    Dansereau, David A.; Lasko, Paul

    2009-01-01

    Summary Germline stem cells (GSCs) in Drosophila are a valuable model to explore of how adult stem cells are regulated in vivo. Genetic dissection of this system has shown that stem cell fate is determined and maintained by the stem cell’s somatic microenvironment or niche. In Drosophila gonads, the stem cell niche—the cap cell cluster in females and the hub in males—acts as a signaling center to recruit GSCs from among a small population of undifferentiated primordial germ cells (PGCs). Short-range signals from the niche specify and regulate stem cell fate by maintaining the undifferentiated state of the PGCs next to the niche. Germline cells that do not receive the niche signals because of their location assume the default fate and differentiate. Once GSCs are specified, adherens junctions maintain close association between the stem cells and their niche and help to orient stem cell division so that one daughter is displaced from the niche and differentiates. In females, stem cell fate depends on bone morphogenetic protein (BMP) signals from the cap cells; in males, hub cells express the cytokine-like ligand Unpaired, which activates the Janus kinase-signal transducers and activators of transcription (Jak-Stat) pathway in stem cells. Although the signaling pathways operating between the niche and stem cells are different, there are common general features in both males and females, including the arrangement of cell types, many of the genes used, and the logic of the system that maintains stem cell fate. PMID:18370048

  5. 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

  6. miR-965 controls cell proliferation and migration during tissue morphogenesis in the Drosophila abdomen.

    PubMed

    Verma, Pushpa; Cohen, Stephen M

    2015-01-01

    Formation of the Drosophila adult abdomen involves a process of tissue replacement in which larval epidermal cells are replaced by adult cells. The progenitors of the adult epidermis are specified during embryogenesis and, unlike the imaginal discs that make up the thoracic and head segments, they remain quiescent during larval development. During pupal development, the abdominal histoblast cells proliferate and migrate to replace the larval epidermis. Here, we provide evidence that the microRNA, miR-965, acts via string and wingless to control histoblast proliferation and migration. Ecdysone signaling downregulates miR-965 at the onset of pupariation, linking activation of the histoblast nests to the hormonal control of metamorphosis. Replacement of the larval epidermis by adult epidermal progenitors involves regulation of both cell-intrinsic events and cell communication. By regulating both cell proliferation and cell migration, miR-965 contributes to the robustness of this morphogenetic system. PMID:26226636

  7. Development and organization of the larval nervous system in Phoronopsis harmeri: new insights into phoronid phylogeny

    PubMed Central

    2014-01-01

    Background The organization and development of the nervous system has traditionally been used as an important character for establishing the relationships among large groups of animals. According to this criterion, phoronids were initially regarded as deuterostomian but have more recently been regarded as protostomian. The resolving of this conflict requires detailed information from poorly investigated members of phoronids, such as Phoronopsis harmeri. Results The serotonin-like immunoreactive part of the P. harmeri nervous system changes during larval development. These changes mostly concern the nervous system of the hood and correlate with the appearance of the median and two marginal neurite bundles, the frontal organ, and the sensory field. The apical organ has bilateral symmetry. The tentacular neurite bundle passes under the tentacles, contains several types of perikarya, and gives rise to intertentacular bundles, which branch in the tentacle base and penetrate into adjacent tentacles by two lateroabfrontal bundles. There are two groups of dorsolateral perikarya, which exhibit serotonin-like immunoreactivity, contact the tentacular neurite bundle, and are located near the youngest tentacles. Larvae have a minor nerve ring, which originates from the posterior marginal neurite bundle of the hood, passes above the tentacle base, and gives rise to the mediofrontal neurite bundle in each tentacle. Paired laterofrontal neurite bundles of tentacles form a continuous nerve tract that conducts to the postoral ciliated band. Discussion The organization of the nervous system differs among the planktotrophic larvae of phoronid species. These differences may correlate with differences in phoronid biology. Data concerning the innervation of tentacles in different phoronid larvae are conflicting and require careful reinvestigation. The overall organization of the nervous system in phoronid larvae has more in common with the deuterostomian than with the protostomian

  8. Developing methods based on light sheet fluorescence microscopy for biophysical investigations of larval zebrafish

    NASA Astrophysics Data System (ADS)

    Taormina, Michael J.

    Adapting the tools of optical microscopy to the large-scale dynamic systems encountered in the development of multicellular organisms provides a path toward understanding the physical processes necessary for complex life to form and function. Obtaining quantitatively meaningful results from such systems has been challenging due to difficulty spanning the spatial and temporal scales representative of the whole, while also observing the many individual members from which complex and collective behavior emerges. A three-dimensional imaging technique known as light sheet fluorescence microscopy provides a number of significant benefits for surmounting these challenges and studying developmental systems. A thin plane of fluorescence excitation light is produced such that it coincides with the focal plane of an imaging system, providing rapid acquisition of optically sectioned images that can be used to construct a three-dimensional rendition of a sample. I discuss the implementation of this technique for use in larva of the model vertebrate Danio rerio (zebrafish). The nature of light sheet imaging makes it especially well suited to the study of large systems while maintaining good spatial resolution and minimizing damage to the specimen from excessive exposure to excitation light. I show the results from a comparative study that demonstrates the ability to image certain developmental processes non-destructively, while in contrast confocal microscopy results in abnormal growth due to phototoxicity. I develop the application of light sheet microscopy to the study of a previously inaccessible system: the bacterial colonization of a host organism. Using the technique, we are able to obtain a survey of the intestinal tract of a larval zebrafish and observe the location of microbes as they grow and establish a stable population in an initially germ free fish. Finally, I describe a new technique to measure the fluid viscosity of this intestinal environment in vivo using

  9. Embryonic multipotent progenitors remodel the Drosophila airways during metamorphosis

    PubMed Central

    Pitsouli, Chrysoula; Perrimon, Norbert

    2010-01-01

    Adult structures in holometabolous insects such as Drosophila are generated by groups of imaginal cells dedicated to the formation of different organs. Imaginal cells are specified in the embryo and remain quiescent until the larval stages, when they proliferate and differentiate to form organs. The Drosophila tracheal system is extensively remodeled during metamorphosis by a small number of airway progenitors. Among these, the spiracular branch tracheoblasts are responsible for the generation of the pupal and adult abdominal airways. To understand the coordination of proliferation and differentiation during organogenesis of tubular organs, we analyzed the remodeling of Drosophila airways during metamorphosis. We show that the embryonic spiracular branch tracheoblasts are multipotent cells that express the homeobox transcription factor Cut, which is necessary for their survival and normal development. They give rise to three distinct cell populations at the end of larval development, which generate the adult tracheal tubes, the spiracle and the epidermis surrounding the spiracle. Our study establishes the series of events that lead to the formation of an adult tubular structure in Drosophila. PMID:20940225

  10. The Impact of Seawater Saturation State on Early Skeletal Development in Larval Corals: Insights into Scleractinian Biomineralization

    NASA Astrophysics Data System (ADS)

    Cohen, A. L.; McCorkle, D. C.; de Putron, S.

    2007-12-01

    contrast to the fine, closed, densely packed spherulitic bundles accreted in the control system, larvae in the lower Omega treatments produced a disorganized conglomerate of large, highly faceted crystals, consistent with slow growth under low saturation state conditions. Our results suggest that the coral calcification response to changes in seawater saturation state is linked to a physiological limitation on the organism's ability to elevate the saturation state of seawater within the calcifying space. Further, our data indicate that ocean acidification due to fossil fuel CO2 emissions will likely have a strong negative effect on the recruitment and early skeletal development of larval corals over the next several decades.

  11. Stage-Specific Changes in Physiological and Life-History Responses to Elevated Temperature and Pco2 during the Larval Development of the European Lobster Homarus gammarus (L.).

    PubMed

    Small, Daniel P; Calosi, Piero; Boothroyd, Dominic; Widdicombe, Steve; Spicer, John I

    2015-01-01

    An organism's physiological processes form the link between its life-history traits and the prevailing environmental conditions, especially in species with complex life cycles. Understanding how these processes respond to changing environmental conditions, thereby affecting organismal development, is critical if we are to predict the biological implications of current and future global climate change. However, much of our knowledge is derived from adults or single developmental stages. Consequently, we investigated the metabolic rate, organic content, carapace mineralization, growth, and survival across each larval stage of the European lobster Homarus gammarus, reared under current and predicted future ocean warming and acidification scenarios. Larvae exhibited stage-specific changes in the temperature sensitivity of their metabolic rate. Elevated Pco2 increased C∶N ratios and interacted with elevated temperature to affect carapace mineralization. These changes were linked to concomitant changes in survivorship and growth, from which it was concluded that bottlenecks were evident during H. gammarus larval development in stages I and IV, the transition phases between the embryonic and pelagic larval stages and between the larval and megalopa stages, respectively. We therefore suggest that natural changes in optimum temperature during ontogeny will be key to larvae survival in a future warmer ocean. The interactions of these natural changes with elevated temperature and Pco2 significantly alter physiological condition and body size of the last larval stage before the transition from a planktonic to a benthic life style. Thus, living and growing in warm, hypercapnic waters could compromise larval lobster growth, development, and recruitment. PMID:26658247

  12. Nitric oxide directly regulates gene expression during Drosophila development: need some gas to drive into metamorphosis?

    PubMed

    Yamanaka, Naoki; O'Connor, Michael B

    2011-07-15

    Nitric oxide (NO) is an important second messenger involved in numerous biological processes, but how it regulates gene expression is not well understood. In this issue of Genes & Development, Cáceres and colleagues (pp. 1476-1485) report a critical requirement of NO as a direct regulator of gene expression through its binding to a heme-containing nuclear receptor in Drosophila. This may be an anciently evolved mechanism to coordinate behavior and metabolism during animal development. PMID:21764850

  13. Effect of benserazid and 6-hydroxydopamine on the development of the last larval instar of the house cricket, Acheta domestica L.

    PubMed

    Rózsa, K S; Chudakova, I V; Hiripi, L

    1986-01-01

    The effect of benserazid and 6-OHDA on the duration of the last larval instar and development of wings with intact corpora allata and following allatectomy on Acheta domestica L. was studied. 6-OHDA failed to alter the duration of the last larval instar in the nymphs with intact corpora allata but prolonged it in the allatectomized crickets. Introduction of 6-OHDA in the second phase of the last larval instar caused a deformation of the wings both in allatectomized and intact crickets. Injection of benserazid to the nymphs of the last larval instar prolonged the duration of this phase only in the presence of the corpora allata. The effect of benserazid can be connected to elimination of central inhibition of the corpora allata involving dopaminergic neurons of the brain. PMID:2869911

  14. Intracellular Trafficking in Drosophila Visual System Development: A Basis for Pattern Formation Through Simple Mechanisms

    PubMed Central

    Chan, Chih-Chiang; Epstein, Daniel; Hiesinger, P. Robin

    2012-01-01

    Intracellular trafficking underlies cellular functions ranging from membrane remodeling to receptor activation. During multicellular organ development, these basic cell biological functions are required as both passive machinery and active signaling regulators. Exocytosis, endocytosis, and recycling of several key signaling receptors have long been known to actively regulate morphogenesis and pattern formation during Drosophila eye development. Hence, intracellular membrane trafficking not only sets the cell biological stage for receptor-mediated signaling but also actively controls signaling through spatiotemporally regulated receptor localization. In contrast to eye development, the role of intracellular trafficking for the establishment of the eye-to-brain connectivity map has only recently received more attention. It is still poorly understood how guidance receptors are spatiotemporally regulated to serve as meaningful synapse formation signals. Yet, the Drosophila visual system provides some of the most striking examples for the regulatory role of intracellular trafficking during multicellular organ development. In this review we will first highlight the experimental and conceptual advances that motivate the study of intracellular trafficking during Drosophila visual system development. We will then illuminate the development of the eye, the eye-to-brain connectivity map and the optic lobe from the perspective of cell biological dynamics. Finally, we provide a conceptual framework that seeks to explain how the interplay of simple genetically encoded intracellular trafficking events governs the seemingly complex cellular behaviors, which in turn determine the developmental product. PMID:21714102

  15. Bazooka mediates secondary axon morphology in Drosophila brain lineages

    PubMed Central

    2011-01-01

    In the Drosophila brain, neural lineages project bundled axon tracts into a central neuropile. Each lineage exhibits a stereotypical branching pattern and trajectory, which distinguish it from other lineages. In this study, we used a multilineage approach to explore the neural function of the Par-complex member Par3/Bazooka in vivo. Drosophila bazooka is expressed in post-mitotic neurons of the larval brain and localizes within neurons in a lineage-dependent manner. The fact that multiple GAL4 drivers have been mapped to several lineages of the Drosophila brain enables investigation of the role of Bazooka from larval to adult stages Bazooka loss-of-function (LOF) clones had abnormal morphologies, including aberrant pathway choice of ventral projection neurons in the BAla1 lineage, ectopic branching in the DALv2 and BAmv1 lineages, and excess BLD5 lineage axon projections in the optic medulla. Exogenous expression of Bazooka protein in BAla1 neurons rescued defective guidance, supporting an intrinsic requirement for Bazooka in the post-mitotic neuron. Elimination of the Par-complex member Par6 recapitulated Bazooka phenotypes in some but not all lineages, suggesting that the Par complex functions in a lineage-dependent manner, and that Bazooka may act independently in some lineages. Importantly, this study highlights the potential of using a multilineage approach when studying gene function during neural development in Drosophila. PMID:21524279

  16. Bazooka mediates secondary axon morphology in Drosophila brain lineages.

    PubMed

    Spindler, Shana R; Hartenstein, Volker

    2011-01-01

    In the Drosophila brain, neural lineages project bundled axon tracts into a central neuropile. Each lineage exhibits a stereotypical branching pattern and trajectory, which distinguish it from other lineages. In this study, we used a multilineage approach to explore the neural function of the Par-complex member Par3/Bazooka in vivo. Drosophila bazooka is expressed in post-mitotic neurons of the larval brain and localizes within neurons in a lineage-dependent manner. The fact that multiple GAL4 drivers have been mapped to several lineages of the Drosophila brain enables investigation of the role of Bazooka from larval to adult stages Bazooka loss-of-function (LOF) clones had abnormal morphologies, including aberrant pathway choice of ventral projection neurons in the BAla1 lineage, ectopic branching in the DALv2 and BAmv1 lineages, and excess BLD5 lineage axon projections in the optic medulla. Exogenous expression of Bazooka protein in BAla1 neurons rescued defective guidance, supporting an intrinsic requirement for Bazooka in the post-mitotic neuron. Elimination of the Par-complex member Par6 recapitulated Bazooka phenotypes in some but not all lineages, suggesting that the Par complex functions in a lineage-dependent manner, and that Bazooka may act independently in some lineages. Importantly, this study highlights the potential of using a multilineage approach when studying gene function during neural development in Drosophila. PMID:21524279

  17. Expression profile of heat shock response factors during hookworm larval activation and parasitic development.

    PubMed

    Gelmedin, Verena; Delaney, Angela; Jennelle, Lucas; Hawdon, John M

    2015-07-01

    protein expression, slightly down regulated both genes under similar conditions. Both modulators inhibited activation-associated feeding, but neither had an effect on hsp-1 levels in activated L3 at 16h. Both celastrol and KNK437 prevent the up-regulation of daf-21 and hsf-1 seen in non-activated control larvae during activation, and significantly down regulated expression of the HSF-1 negative regulator Aca-hsb-1 in activated larvae. Expression levels of heat shock response factors were examined in developing Ancylostoma ceylanicum larvae recovered from infected hosts and found to differ significantly from the expression profile of activated L3, suggesting that feeding during in vitro activation is regulated differently than parasitic development. Our results indicate that a classical heat shock response is not induced at host temperature and is suppressed during larval recovery and parasitic development in the host, but a partial heat shock response is induced after extended incubation at host temperature in the absence of a developmental signal, possibly to protect against heat stress. PMID:26296769

  18. Experimental study of Lucilia sericata (Diptera Calliphoridae) larval development on rat cadavers: Effects of climate and chemical contamination.

    PubMed

    Aubernon, Cindy; Charabidzé, Damien; Devigne, Cédric; Delannoy, Yann; Gosset, Didier

    2015-08-01

    Household products such as bleach, gasoline or hydrochloric acid have been used to mask the presence of a cadaver or to prevent the colonization of insects. These types of chemicals affect insect development and alter the forensic entomology analysis. This study was designed to test the effects of six household products (bleach, mosquito repellent, perfume, caustic soda, insecticide and unleaded gasoline) on blowfly (Lucilia sericata, Diptera: Calliphoridae) larval development. Furthermore, the effects of climate (rain or dry conditions) on larval development were analyzed. For each replication, 100 first instars were placed on a rat cadaver on which one household product was spilled. We observed a decrease in the survival rates of the larvae but no significant effect on their development times or the adult size. The same trends were observed under rainy conditions. However, the rain altered the effects of some tested household products, especially gasoline. These results demonstrate for the first time the successful development of necrophagous larvae on chemically contaminated cadavers, and provide evidence for the range of possible effects to expect. PMID:26123620

  19. The vascular anatomy of the developing zebrafish: an atlas of embryonic and early larval development.

    PubMed

    Isogai, S; Horiguchi, M; Weinstein, B M

    2001-02-15

    We have used confocal microangiography to examine and describe the vascular anatomy of the developing zebrafish, Danio rerio. This method and the profound optical clarity of zebrafish embryos make it possible to view the entire developing vasculature with unprecedented resolution. A staged series of three-dimensional images of the vascular system were collected beginning shortly after the onset of circulation at 1 day postfertilization through early- to midlarval stages at approximately 7 days postfertilization. Blood vessels in every region of the animal were imaged at each stage, and detailed "wiring patterns" were derived describing the interconnections between every major vessel. We present an overview of these data here in this paper and in an accompanying Web site "The interactive atlas of zebrafish vascular anatomy" online at (http://eclipse.nichd.nih.gov/nichd/lmg/redirect.html). We find a highly dynamic but also highly stereotypic pattern of vascular connections, with different sets of primitive embryonic vessels severing connections and rewiring in new configurations according to a reproducible plan. We also find that despite variation in the details of the vascular anatomy, the basic vascular plan of the developing zebrafish shows strong similarity to that of other vertebrates. This atlas will provide an invaluable foundation for future genetic and experimental studies of vascular development in the zebrafish. PMID:11161578

  20. The effect of space environment on the development and aging of Drosophila Melanogaster (7-IML-1)

    NASA Technical Reports Server (NTRS)

    Marco, Roberto

    1992-01-01

    This experiment involves the study of the development of eggs of the fly, Drosophila, exposed to microgravity. It is presumed that oogenesis, rather than further states of embryonic development, is sensitive to gravity. This hypothesis will be tested by collecting eggs layered at specific times inflight and postflight from flies exposed to 0 and 1 g. This portion of the experiment is a repetition of an earlier experiment flown in Biorack during the Spacelab D1 Mission. An added feature of the experiment for the First International Microgravity Laboratory (IML-1) Mission is to study the effect of microgravity on the life span of Drosophila male flies. Various aspects of the investigation are discussed.

  1. Effect of mercuric chloride on fertilization and larval development in the River Frog, Rana heckscheri (Wright) (Anura: Ranidae)

    SciTech Connect

    Punzo, F. )

    1993-10-01

    Previous investigations have indicated that heavy metals such as copper, cadmium, lead and mercury can act as systemic toxicants in many species of wildlife. Although numerous studies have emphasized the effects of metals and pesticides on metabolism, growth, survivorship, neural processes and reproduction in a number of taxa, little information is available on the effects of sublethal concentrations of metals on the reproductive physiology of amphibians. Industrial processes and mining activities can release substantial concentrations of heavy metals such as mercury into aquatic habitats. Since most amphibians have obligate aquatic larval stages, they are exposed to pollutants discharged into the aquatic environment. Amphibians can act as accumulators of heavy metals and their larval stages are useful indicators of pollution levels in the field. What little data are available, indicate that metals can significantly reduce viability in amphibians through their actions on metabolism, development and gametogenesis. The recent concerns over worldwide declines in amphibian populations and the susceptibility of amphibian populations to environmental toxicants, led me to assess the effect of mercuric chloride, one of the most common and persistent toxicants in aquatic environments, on fertilization and larval development in the river frog, Rana heckscheri (Wright). Although there is some information on fish, very little data are available on the effects of mercury on fertilization in amphibians generally, and no published data exist for R. heckscheri. This species is a conspicuous component of the aquatic fauna of parts of the southeastern United States where mercury levels have increased significantly over the last two decades. 22 refs., 2 tabs.

  2. Wnt4 is required for ostia development in the Drosophila heart.

    PubMed

    Chen, Zhimin; Zhu, Jun-Yi; Fu, Yulong; Richman, Adam; Han, Zhe

    2016-05-15

    The Drosophila ostia are valve-like structures in the heart with functional similarity to vertebrate cardiac valves. The Wnt/β-catenin signaling pathway is critical for valve development in zebrafish and mouse, but the key ligand(s) for valve induction remains unclear. We observed high levels of Wnt4 gene expression in Drosophila ostia progenitor cells, immediately prior to morphological differentiation of these cells associated with ostia formation. This differentiation was blocked in Wnt4 mutants and in flies expressing canonical Wnt signaling pathway inhibitors but not inhibitors of the planar cell polarity pathway. High levels of Wnt4 dependent activation of a canonical Wnt signaling reporter was observed specifically in ostia progenitor cells. In vertebrate valve formation Wnt signaling is active in cells undergoing early endothelial-mesenchymal transition (EMT) and the Wnt9 homolog of Drosophila Wnt4 is expressed in valve progenitors. In demonstrating an essential role for Wnt4 in ostia development we have identified similarities between molecular and cellular events associated with early EMT during vertebrate valve development and the differentiation and partial delamination of ostia progenitor cells in the process of ostia formation. PMID:26994311

  3. Overcrowding-mediated stress alters cell proliferation in key neuroendocrine areas during larval development in Rhinella arenarum.

    PubMed

    Distler, Mijal J; Jungblut, Lucas D; Ceballos, Nora R; Paz, Dante A; Pozzi, Andrea G

    2016-02-01

    Exposure to adverse environmental conditions can elicit a stress response, which results in an increase in endogenous corticosterone levels. In early life stages, it has been thoroughly demonstrated that amphibian larval growth and development is altered as a consequence of chronic stress by interfering with the metamorphic process, however, the underlying mechanisms involved have only been partially disentangled. We examined the effect of intraspecific competition on corticosterone levels during larval development of the toad Rhinella arenarum and its ultimate effects on cell proliferation in particular brain areas as well as the pituitary gland. While overcrowding altered the number of proliferating cells in the pituitary gland, hypothalamus, and third ventricle of the brain, no differences were observed in areas which are less associated with neuroendocrine processes, such as the first ventricle of the brain. Apoptosis was increased in hypothalamic regions but not in the pituitary. With regards to pituitary cell populations, thyrotrophs but not somatoatrophs and corticotrophs showed a decrease in the cell number in overcrowded larvae. Our study shows that alterations in growth and development, produced by stress, results from an imbalance in the neuroendocrine systems implicated in orchestrating the timing of metamorphosis. J. Exp. Zool. 9999A:XX-XX, 2016. © 2016 Wiley Periodicals, Inc. PMID:26817921

  4. Patterns of Cranial Development in Larval Rana macrocnemis: Chondrocranial Size and Shape Relationship With Pelophylax bedriagae (Anura: Ranidae).

    PubMed

    Yildirim, Elıf; Kaya, Uğur

    2016-06-01

    Notwithstanding the abundance of amphibians, there are few descriptions about ranid cranial development. Herein, larval chondrocranial development of Uludağ frog, Rana macrocnemis (Boulenger, 1885), is described on cleared and double-stained specimens. Descriptions are related with the ontogeny of the chondrocranium and osteogenesis of the cranial skeleton. The larval chondrocranial development of R. macrocnemis is compared to those of Rana and Pelophylax larvae (Pelophylax bedriagae, Rana pipiens, R. palustris, R. sphenocephala, R. catesbeiana, R. clamitans and R. sylvatica). In R. macrocnemis, the first bones to ossify are the parasphenoid and exoccipital (Stage 33), followed by the frontoparietal and prootic (stages 35 and 40, respectively). The major reconstruction of the chondrocranium begins at Stage 41. The ossification sequence of R. macrocnemis is distinguished from other ranids. Adult cranial osteology of R. macrocnemis is compared to that of P. bedriagae. Osteologically, R. macrocnemis is different from P. bedriagae by the shape and size of the vomer and number of teeth. Additionally, geometric morphometric methods are used to analyze chondrocranial size and shape changes of ranid larva of R. macrocnemis and P. bedriagae. Anat Rec, 299:711-721, 2016. © 2016 Wiley Periodicals, Inc. PMID:26950267

  5. Chronic effects of strobilurin fungicides on development, growth, and mortality of larval Great Plains toads (Bufo cognatus).

    PubMed

    Hartman, Emily A H; Belden, Jason B; Smith, Loren M; McMurry, Scott T

    2014-04-01

    Agricultural fungicide application has increased tenfold since 2005 in the United States. Active ingredients and formulations of strobilurin fungicides at environmentally relevant concentrations cause mortality to larval and metamorph amphibians; however, little is known about chronic exposure effects in amphibians. We exposed larval amphibians (Bufo cognatus) throughout metamorphosis to the common fungicide formulations Headline(®), Stratego(®), Quilt(®), and a control to determine effects on development and growth. Formulations were tested at 1.7, 50, and 400 μg/L of the active strobilurin ingredient for Headline(®), Stratego(®), and Quilt(®), respectively. Fungicide exposure did not affect body mass or snout-vent length at metamorphosis. However, exposure to Headline(®) at 1.7 μg/L increased the development rate of tadpoles by approximately 5 days compared to the control, an effect not observed for Stratego(®) and Quilt(®). Stratego(®) also caused approximately 35 % cumulative mortality. Results from the experiment suggest that chronic effects of strobilurin fungicides on development, growth, and mortality to B. cognatus are apparent at environmentally relevant concentrations. PMID:24522617

  6. Combined effects of pollutants and salinity on embryo-larval development of the Pacific oyster, Crassostrea gigas.

    PubMed

    Gamain, Perrine; Gonzalez, Patrice; Cachot, Jérôme; Pardon, Patrick; Tapie, Nathalie; Gourves, Pierre Yves; Budzinski, Hélène; Morin, Bénédicte

    2016-02-01

    For several years, low larval recruitment has been observed in Arcachon Bay, in southwest France. Exposure to pollutants could partly account for the reduction of early life stages of the Pacific oyster. This study evaluated the effects of copper and S-metolachlor in combination with salinity on the early life stages of Crassostrea gigas. Embryos were exposed to concentrations of copper (1, 10 and 50 μg L(-1)) or S-metolachlor (10, 100 and 1000 ng L(-1)) and six salinities (18, 21, 24, 27, 30 and 33 u.s.i). Embryotoxicity was measured by considering both the percentage of abnormalities and arrested development in D-shaped larvae. Embryo-larval development was only affected at salinities ≤24 u.s.i, which have never been observed during C. gigas reproduction period in Arcachon Bay. Both contaminants had an effect at environmental concentrations. Our results suggest that copper and metolachlor toxicity was enhanced with decreasing salinity. PMID:26583531

  7. The effects of the amphibian chytrid fungus, insecticide exposure, and temperature on larval anuran development and survival.

    PubMed

    Rumschlag, Samantha L; Boone, Michelle D; Fellers, Gary

    2014-11-01

    Chytridiomycosis, a disease caused by Batrachochytrium dendrobatidis (Bd), has been implicated as a cause of amphibian declines. Susceptibility may be influenced by environmental factors that suppress the immune response. The authors conducted a laboratory study to examine the effect of temperature, insecticide exposure, and Bd exposure during larval anuran development. The authors examined the consequences of exposure to Bd, an insecticide (carbaryl or malathion), and static or fluctuating temperature (15 °C, 20 °C, 25 °C, or 15 °C to 25 °C 72-h flux) on larval development through metamorphosis of the Pacific treefrog (Pseudacris regilla). High and fluctuating temperature had negative effects on survival in the presence of Bd. Insecticides inhibited the effects of Bd; time to tail resorption of Pacific treefrogs decreased when tadpoles were exposed to carbaryl. The present study indicates that abiotic factors may play a role in the host-pathogen interactions in this system. PMID:25098758

  8. A novel tissue in an established model system: the Drosophila pupal midgut.

    PubMed

    Takashima, Shigeo; Younossi-Hartenstein, Amelia; Ortiz, Paola A; Hartenstein, Volker

    2011-06-01

    The Drosophila larval and adult midguts are derived from two populations of endodermal progenitors that separate from each other in the early embryo. As larval midgut cells differentiate into an epithelial layer, adult midgut progenitors (AMPs) remain as small clusters of proliferating, undifferentiated cells attached to the basal surface of the larval gut epithelium. During the first few hours of metamorphosis, AMPs merge into a continuous epithelial tube that overgrows the larval layer and differentiates into the adult midgut; at the same time, the larval midgut degenerates. As shown in this paper, there is a second, transient pupal midgut that develops from the AMPs at the beginning of metamorphosis and that intercalates between the adult and larval midgut epithelia. Cells of the transient pupal midgut form a multilayered tube that exhibits signs of differentiation, in the form of septate junctions and rudimentary apical microvilli. Some cells of the pupal midgut develop as endocrine cells. The pupal midgut remains closely attached to the degenerating larval midgut cells. Along with these cells, pupal midgut cells are sequestered into the lumen where they form the compact "yellow body." The formation of a pupal midgut has been reported from several other species and may represent a general feature of intestinal metamorphosis in insects. PMID:21556856

  9. A novel tissue in an established model system: the Drosophila pupal midgut

    PubMed Central

    Takashima, Shigeo; Younossi-Hartenstein, Amelia; Ortiz, Paola A.; Hartenstein, Volker

    2014-01-01

    The Drosophila larval and adult midgut are derived from two populations of endodermal progenitors that separate from each other in the early embryo. As larval midgut cells differentiate into an epithelial layer, adult midgut progenitors (AMPs) remain as small clusters of proliferating, undifferentiated cells attached to the basal surface of the larval gut epithelium. During the first few hours of metamorphosis, AMPs merge into a continuous epithelial tube that overgrows the larval layer and differentiates into the adult midgut; at the same time, the larval midgut degenerates. As shown in this paper, there is a second, transient pupal midgut that develops from the AMPs at the beginning of metamorphosis, and that intercalates between the adult and larval midgut epithelia. Cells of the transient pupal midgut form a multilayered tube that exhibits signs of differentiation, in the form of septate junctions and rudimentary apical microvilli. Some cells of the pupal midgut develop as endocrine cells. The pupal midgut remains closely attached to the degenerating larval midgut cells. Along with these cells, pupal midgut cells are sequestered into the lumen where they form the compact “yellow body”. The formation of a pupal midgut has been reported from several other species, and may represent a general feature of intestinal metamorphosis in insects. PMID:21556856

  10. Larval Defense against Attack from Parasitoid Wasps Requires Nociceptive Neurons

    PubMed Central

    Robertson, Jessica L.; Tsubouchi, Asako; Tracey, W. Daniel

    2013-01-01

    Parasitoid wasps are a fierce predator of Drosophila larvae. Female Leptopilina boulardi (LB) wasps use a sharp ovipositor to inject eggs into the bodies of Drosophila melanogaster larvae. The wasp then eats the Drosophila larva alive from the inside, and an adult wasp ecloses from the Drosophila pupal case instead of a fly. However, the Drosophila larvae are not defenseless as they may resist the attack of the wasps through somatosensory-triggered behavioral responses. Here we describe the full range of behaviors performed by the larval prey in immediate response to attacks by the wasps. Our results suggest that Drosophila larvae primarily sense the wasps using their mechanosensory systems. The range of behavioral responses included both “gentle touch” like responses as well as nociceptive responses. We found that the precise larval response depended on both the somatotopic location of the attack, and whether or not the larval cuticle was successfully penetrated during the course of the attack. Interestingly, nociceptive responses are more likely to be triggered by attacks in which the cuticle had been successfully penetrated by the wasp. Finally, we found that the class IV neurons, which are necessary for mechanical nociception, were also necessary for a nociceptive response to wasp attacks. Thus, the class IV neurons allow for a nociceptive behavioral response to a naturally occurring predator of Drosophila. PMID:24205297

  11. Investigating the effects of nanoparticles on reproduction and development in Drosophila melanogaster and CD-1 mice

    NASA Astrophysics Data System (ADS)

    Philbrook, Nicola Anne

    Manufactured nanoparticles (NPs) are a class of small (≤ 100 nm) materials that are being used for a variety of purposes, including industrial lubricants, food additives, antibacterial agents, as well as delivery systems for drug and gene therapies. Their unique characteristics due to their small size as well as their parent materials allow them to be exploited in convenience applications; however, some of these properties also allow them to interact with and invade biological systems. Few studies have been performed to determine the potential harm that NPs can inflict on reproductive and developmental processes in organisms. In this study, Drosophila melanogaster and CD-1 mice were orally exposed to varying doses of titanium dioxide (TiO 2) NPs, silver (Ag) NPs, or hydroxyl-functionalized carbon nanotubes (fCNTs) and Drosophila were also exposed to microparticles (MPs) as a control for particle size. The subsequent effect of these materials on reproduction and development were evaluated. Strikingly, each type of NP studied negatively affected either reproduction or development in one or both of the two model systems. TiO2 NPs significantly negative effected both CD-1 mouse development (100 mg/kg or 1000 mg/kg) as well as Drosophila female fecundity (0.005%-0.5% w/v). Ag NPs significantly reduced mouse fetus viability after prenatal exposure to10 mg/kg. Ag NPs also significantly decreased the developmental success of Drosophila when they were directly exposed to these NPs (0.05% - 0.5% w/v) compared to both the vehicle and MP controls. fCNTs significantly increased the presence of morphological defects, resorptions and skeletal abnormalities in CD-1 mice, but had little effect on Drosophila. We speculate that the differences seen in the effects of NP types may be partially due to differences in reproductive physiology as well as each organism's ability to internalize these NPs. Whereas the differing response of each organism to a NP type was likely due in part to

  12. Drosophila Cyclin G Is a Regulator of the Notch Signalling Pathway during Wing Development

    PubMed Central

    Nagel, Anja C.; Szawinski, Jutta; Zimmermann, Mirjam; Preiss, Anette

    2016-01-01

    Notch signalling regulates a multitude of differentiation processes during Drosophila development. For example, Notch activity is required for proper wing vein differentiation which is hampered in mutants of either the receptor Notch, the ligand Delta or the antagonist Hairless. Moreover, the Notch pathway is involved in several aspects of Drosophila oogenesis as well. We have identified Drosophila Cyclin G (CycG) as a molecular interaction partner of Hairless, the major antagonist in the Notch signalling pathway, in vitro and in vivo. Loss of CycG was shown before to cause female sterility and to disturb the architecture of the egg shell. Nevertheless, Notch dependent processes during oogenesis appeared largely unaffected in cycG mutant egg chambers. Loss of CycG modified the dominant wing phenotypes of Notch, Delta and Hairless mutants. Whereas the Notch loss of function phenotype was ameliorated by a loss of CycG, the phenotypes of either Notch gain of function or of Delta or Hairless loss of function were enhanced. In contrast, loss of CycG had only a minor effect on the wing vein phenotype of mutants affecting the EGFR signalling pathway emphasizing the specificity of the interaction of CycG and Notch pathway members. PMID:26963612

  13. Microgravity effects on the oogenesis and development of embryos of Drosophila melanogaster laid in the Spaceshuttle during the Biorack experiment (ESA).

    PubMed

    Vernós, I; González-Jurado, J; Calleja, M; Marco, R

    1989-06-01

    The results obtained during the last successful flight of the Challenger Shuttle, in early November 1985, indicate that oogenesis and embryonic development of Drosophila melanogaster are altered in the absence of gravity. Two hundred forty females and ninety males, wild type Oregon R Drosophila melanogaster flies were flown in the Spaceshuttle during the 7-day D-1 mission and the embryos laid during the spaceflight were recovered and studied. Although some eggs developed into normal 1st instar larvae and many into late embryos in the 23 +/- 2 h collection periods throughout the flight, several interesting differences from the parallel ground and in-flight centrifuge controls were observed: 1) There was an increase in oocyte production and size. 2) There was a significant decrease in the number of larvae hatched from the embryonic cuticles in microgravity. 3) The majority of embryos were normally fertilized and at late stages of development, except in the space-flown containers in microgravity where a percentage of earlier stage embryos were recovered showing alterations in the deposition of yolk. 4) In correspondence with these results, at least 25% of the living embryos recovered from space failed to develop into adults. 5) Studies of the larval cuticles and those of the late embryos indicate the existence of alterations in the anterior, head and thoracic regions of the animals. 6) There was a delay in the development into adults of the embryos and larvae that had been subjected to microgravity and recovered from the space shuttle at the end of the flight. No significant accumulation of lethal mutations in any of the experimental conditions was detected as measured through the male to female ratio in the descendant generation. It seems that Drosophila melanogaster flies are able to sense and respond to the absence of gravity, changing several developmental processes even in very short space flights. The results suggest an interference with the distribution and

  14. Larval development of the Chinese mitten crab Eriocheir sinensis H. Milne-Edwards (Decapoda: Grapsidae) reared in the laboratory

    NASA Astrophysics Data System (ADS)

    Montú, M.; Anger, K.; de Bakker, C.

    1996-06-01

    Larvae of the Chinese mitten crab Eriocheir sinensis were reared in the laboratory from the time of hatching and through metamorphosis. Development normally consists of a Prezoea, 5 Zoea stages, and a Megalopa. Occasionally, an additional (stage VI) Zoea and, in one case, an additional Megalopa (transitional to the first crab stage) were observed. Detailed morphological descriptions of all larval and the first two juvenile instars are given, and larval morphology is compared with that of two closely related species, Eriocheir japonicus and Eriocheir rectus, descriptions of which have recently become available. The zoeal stages of these species can be distinguished by their different number of aesthetascs and setae on the antennules, and different setation of maxillipeds 1 and 2. The Megalopa shows differences in the shape of the rostrum and again in the morphology of the antennule. These and other morphological differences (mainly in setation and spinulation of the zoeal carapace) between E. sinensis and E. japonicus larvae suggest that they may be very closely related but separate species; this contradicts a recent study of adult morphometrics and molecular genetics (Li et al., 1993), suggesting that they are only varieties of a single species.

  15. Embryonic, Larval, and Juvenile Development of the Sea Biscuit Clypeaster subdepressus (Echinodermata: Clypeasteroida)

    PubMed Central

    Vellutini, Bruno C.; Migotto, Alvaro E.

    2010-01-01

    Sea biscuits and sand dollars diverged from other irregular echinoids approximately 55 million years ago and rapidly dispersed to oceans worldwide. A series of morphological changes were associated with the occupation of sand beds such as flattening of the body, shortening of primary spines, multiplication of podia, and retention of the lantern of Aristotle into adulthood. To investigate the developmental basis of such morphological changes we documented the ontogeny of Clypeaster subdepressus. We obtained gametes from adult specimens by KCl injection and raised the embryos at 26C. Ciliated blastulae hatched 7.5 h after sperm entry. During gastrulation the archenteron elongated continuously while ectodermal red-pigmented cells migrated synchronously to the apical plate. Pluteus larvae began to feed in 3 d and were 20 d old at metamorphosis; starved larvae died 17 d after fertilization. Postlarval juveniles had neither mouth nor anus nor plates on the aboral side, except for the remnants of larval spicules, but their bilateral symmetry became evident after the resorption of larval tissues. Ossicles of the lantern were present and organized in 5 groups. Each group had 1 tooth, 2 demipyramids, and 2 epiphyses with a rotula in between. Early appendages consisted of 15 spines, 15 podia (2 types), and 5 sphaeridia. Podial types were distributed in accordance to Lovén's rule and the first podium of each ambulacrum was not encircled by the skeleton. Seven days after metamorphosis juveniles began to feed by rasping sand grains with the lantern. Juveniles survived in laboratory cultures for 9 months and died with wide, a single open sphaeridium per ambulacrum, aboral anus, and no differentiated food grooves or petaloids. Tracking the morphogenesis of early juveniles is a necessary step to elucidate the developmental mechanisms of echinoid growth and important groundwork to clarify homologies between irregular urchins. PMID:20339592

  16. [Embrionary-larval development of the tropical fish Hemirhamphus brasiliensis (Beloniformes: Hemirhamphidae) from eggs collected in the wild].

    PubMed

    Rosas, Jesús; Mata, Ernesto; Velásquez, Aidé; Cabrera, Tomas

    2008-09-01

    The embryo formation and larval development of Hemirhamphus brasiliensis Linnaeus, 1758 (Pisces: Hemirhamphidae) is described from morula stage eggs collected on Sargassum sp. Thalii in the field (10 degrees 50'55.2" N y 64 degrees 09'467" W). The eggs were spherical, 1 923.54 +/- 72.35 microm diameter with several corionic filaments, and are striated. During the first 48 h the embryo developed cephalic vesicle, miomers, and a heart located on the external body surface, beating strongly and circulating colorless blood which became pigmented red later. Before hatching, the larva developed kidney, gut tract, liver and biliar vesicle, pectoral fins, four pairs of gill arches and the mouth. The larva hatched at 114 h, the body was torpedo-shaped, yellow-green, with several dendriform melanophores; the pelvic fin was observed 72 h post hatching. At 240 hours the metamorphoses was completed. When the larvae hatched they could ingest Artemia metanauplii. PMID:19419056

  17. 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

  18. Isolation of intact larval haemoglobin from the brine shrimp Artemia salina. Prevention of degradation in vitro by proteases induced during larval development.

    PubMed

    Mansfield, B C; Krissansen, G W; Smith, M G; Tate, W P

    1980-05-29

    Haemoglobin induced in the larval stage of the brine shrimp, Artemia salina is extensively degraded when isolated from the later developmental stages of the larvae. Alkaline proteases appear in the organism a few hours after the induction of haemoglobin and cause the observed degradation. Addition of 2.6 mM phenylmethylsulphonyl fluoride or 20 micrograms/ml soybean trypsin inhibitor to the extraction buffer used for haemoglobin isolation prevents most of this degradation. Discrete haem proteins are found in extracts of the brine shrimp larvae isolated before induction of the proteases, and the major species has a molecular weight of over 200,000. This is believed to be the native haemoglobin. A spread of lower molecular weight haem-containing polypeptides is found in extracts of larvae isolated after induction of the proteases. These products are believed to result from degradation of the discrete haem proteins present in protease-free extracts. PMID:6990994

  19. Importin-β11 Regulates Synaptic pMAD and Thereby Influences Synaptic Development and Function at the Drosophila Neuromuscular Junction

    PubMed Central

    Higashi-Kovtun, Misao E.; Mosca, Timothy J.; Dickman, Dion K.; Meinertzhagen, Ian A.; Schwarz, Thomas L.

    2010-01-01

    Importin proteins act both at the nuclear pore to promote substrate entry and in the cytosol during signal trafficking. Here, we describe mutations in the Drosophila gene importin-β11 which has not previously been analyzed genetically. Mutants of importin-β11 died as late pupae from neuronal defects and neuronal importin-β11 was present not only at nuclear pores but also in the cytosol and at synapses. Neurons lacking importin-β11 were viable and properly differentiated but exhibited discrete defects. Synaptic transmission was defective in adult photoreceptors and at larval neuromuscular junctions. Mutant photoreceptor axons formed grossly normal projections and synaptic terminals in the brain, but synaptic arbors on larval muscles were smaller while still containing appropriate synaptic components. BMP signaling was the apparent cause of the observed NMJ defects. Importin-β11 interacted genetically with the BMP pathway and at mutant synaptic boutons, a key component of this pathway, phosphorylated Mothers Against Decapentaplegic (pMAD), was reduced. Neuronal expression of an importin-β11 transgene rescued this phenotype as well as the other observed neuromuscular phenotypes. Despite the loss of synaptic pMAD, pMAD persisted in motor neuron nuclei, suggesting a specific impairment in the local function of pMAD. Restoring levels of pMAD to mutant terminals via expression of constitutively active type I BMP receptors or by reducing retrograde transport in motor neurons, also restored synaptic strength and morphology. Thus, importin-β11 function interacts with the BMP pathway to regulate a pool of pMAD that must be present at the presynapse for its proper development and function. PMID:20392948

  20. Foraging Path-length Protocol for Drosophila melanogaster Larvae.

    PubMed

    Anreiter, Ina; Vasquez, Oscar E; Allen, Aaron M; Sokolowski, Marla B

    2016-01-01

    The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral variation. The larval path-length assay was developed to measure individual differences in foraging behavior that were later linked to the foraging gene. Larval path-length is an easily scored trait that facilitates the collection of large sample sizes, at minimal cost, for genetic screens. Here we provide a detailed description of the current protocol for the larval path-length assay first used by Sokolowski. The protocol details how to reproducibly handle test animals, perform the behavioral assay and analyze the data. An example of how the assay can be used to measure behavioral plasticity in response to environmental change, by manipulating feeding environment prior to performing the assay, is also provided. Finally, appropriate test design as well as environmental factors that can modify larval path-length such as food quality, developmental age and day effects are discussed. PMID:27167330

  1. The role of Dichaete in transcriptional regulation during Drosophila embryonic development

    PubMed Central

    2013-01-01

    Background Group B Sox domain transcription factors play conserved roles in the specification and development of the nervous system in higher metazoans. However, we know comparatively little about how these transcription factors regulate gene expression, and the analysis of Sox gene function in vertebrates is confounded by functional compensation between three closely related family members. In Drosophila, only two group B Sox genes, Dichaete and SoxN, have been shown to function during embryonic CNS development, providing a simpler system for understanding the functions of this important class of regulators. Results Using a combination of transcriptional profiling and genome-wide binding analysis we conservatively identify over 1000 high confidence direct Dichaete target genes in the Drosophila genome. We show that Dichaete plays key roles in CNS development, regulating aspects of the temporal transcription factor sequence that confer neuroblast identity. Dichaete also shows a complex interaction with Prospero in the pathway controlling the switch from stem cell self-renewal to neural differentiation. Dichaete potentially regulates many more genes in the Drosophila genome and was found to be associated with over 2000 mapped regulatory elements. Conclusions Our analysis suggests that Dichaete acts as a transcriptional hub, controlling multiple regulatory pathways during CNS development. These include a set of core CNS expressed genes that are also bound by the related Sox2 gene during mammalian CNS development. Furthermore, we identify Dichaete as one of the transcription factors involved in the neural stem cell transcriptional network, with evidence supporting the view that Dichaete is involved in controlling the temporal series of divisions regulating neuroblast identity. PMID:24314314

  2. 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...

  3. Differential toxicity of silver and titanium dioxide nanoparticles on Drosophila melanogaster development, reproductive effort, and viability: size, coatings and antioxidants matter.

    PubMed

    Posgai, Ryan; Cipolla-McCulloch, Caitlin B; Murphy, Kyle R; Hussain, Saber M; Rowe, John J; Nielsen, Mark G

    2011-09-01

    Silver and titanium dioxide nanoparticles are known to induce oxidative stress in vitro and in vivo. Here we test if they impact development, mating success, and survivorship in Drosophila melanogaster, and if so, if these effects are reversible by antioxidants. Ingestion of nanotitanium dioxide during the larval stage of the life cycle showed no effects on development or survivorship, up to doses of 200 μg mL(-1). Conversely, ingestion of nanosilver had major dose, size, and coating-dependent effects on each of these aspects of life history. Each of these effects was partially or fully reversible by vitamin C. Larvae growing on nanosilver supplemented with vitamin C showed a greater than twofold increase in survivorship compared to flies reared on nanosilver alone, and a threefold increase in mating success. Vitamin C also rescued cuticular and pigmentation defects in nanosilver fed flies. Biochemical assays of superoxide dismutase and glutathione show these markers respond to nanotitanium dioxide and nanosilver induced oxidative stress, and this response is reduced by vitamin C. These results indicate that life history effects of nanosilver ingestion result from oxidative stress, and suggest antioxidants as a potential remediation for nanosilver toxicity. Conversely, the lack of nanotitanium dioxide life history toxicity shows that oxidative stress does not necessarily result in whole organism effects, and argues that nanoparticle toxicity needs to be examined at different levels of biological organization. PMID:21733543

  4. Looming detection by identified visual interneurons during larval development of the locust Locusta migratoria.

    PubMed

    Simmons, Peter J; Sztarker, Julieta; Rind, F Claire

    2013-06-15

    Insect larvae clearly react to visual stimuli, but the ability of any visual neuron in a newly hatched insect to respond selectively to particular stimuli has not been directly tested. We characterised a pair of neurons in locust larvae that have been extensively studied in adults, where they are known to respond selectively to objects approaching on a collision course: the lobula giant motion detector (LGMD) and its postsynaptic partner, the descending contralateral motion detector (DCMD). Our physiological recordings of DCMD axon spikes reveal that at the time of hatching, the neurons already respond selectively to objects approaching the locust and they discriminate between stimulus approach speeds with differences in spike frequency. For a particular approaching stimulus, both the number and peak frequency of spikes increase with instar. In contrast, the number of spikes in responses to receding stimuli decreases with instar, so performance in discriminating approaching from receding stimuli improves as the locust goes through successive moults. In all instars, visual movement over one part of the visual field suppresses a response to movement over another part. Electron microscopy demonstrates that the anatomical substrate for the selective response to approaching stimuli is present in all larval instars: small neuronal processes carrying information from the eye make synapses both onto LGMD dendrites and with each other, providing pathways for lateral inhibition that shape selectivity for approaching objects. PMID:23531812

  5. Mitochondrial fusion but not fission regulates larval growth and synaptic development through steroid hormone production

    PubMed Central

    Sandoval, Hector; Yao, Chi-Kuang; Chen, Kuchuan; Jaiswal, Manish; Donti, Taraka; Lin, Yong Qi; Bayat, Vafa; Xiong, Bo; Zhang, Ke; David, Gabriela; Charng, Wu-Lin; Yamamoto, Shinya; Duraine, Lita; Graham, Brett H; Bellen, Hugo J

    2014-01-01

    Mitochondrial fusion and fission affect the distribution and quality control of mitochondria. We show that Marf (Mitochondrial associated regulatory factor), is required for mitochondrial fusion and transport in long axons. Moreover, loss of Marf leads to a severe depletion of mitochondria in neuromuscular junctions (NMJs). Marf mutants also fail to maintain proper synaptic transmission at NMJs upon repetitive stimulation, similar to Drp1 fission mutants. However, unlike Drp1, loss of Marf leads to NMJ morphology defects and extended larval lifespan. Marf is required to form contacts between the endoplasmic reticulum and/or lipid droplets (LDs) and for proper storage of cholesterol and ecdysone synthesis in ring glands. Interestingly, human Mitofusin-2 rescues the loss of LD but both Mitofusin-1 and Mitofusin-2 are required for steroid-hormone synthesis. Our data show that Marf and Mitofusins share an evolutionarily conserved role in mitochondrial transport, cholesterol ester storage and steroid-hormone synthesis. DOI: http://dx.doi.org/10.7554/eLife.03558.001 PMID:25313867

  6. Post-embryonic larval development and metamorphosis of the hydroid Eudendrium racemosum (Cavolini) (Hydrozoa, Cnidaria)

    NASA Astrophysics Data System (ADS)

    Sommer, C.

    1990-09-01

    The morphology and histology of the planula larva of Eudendrium racemosum (Cavolini) and its metamorphosis into the primary polyp are described from light microscopic observations. The planula hatches as a differentiated gastrula. During the lecithotrophic larval period, large ectodermal mucous cells, embedded between epitheliomuscular cells, secrete a sticky slime. Two granulated cell types occur in the ectoderm that are interpreted as secretory and sensorynervous cells, but might also be representatives of only one cell type with a multiple function. The entoderm consists of yolk-storing gastrodermal cells, digestive gland cells, interstitial cells, cnidoblasts, and premature cnidocytes. The larva starts metamorphosis by affixing its blunt aboral pole to a substratum. While the planula flattens down, the mucous cells penetrate the mesolamella and migrate through the entoderm into the gastral cavity where they are lysed. Subsequently, interstitial cells, cnidoblasts, and premature cnidocytes migrate in the opposite direction, i.e. from entoderm to ectoderm. Then, the polypoid body organization, comprising head (hydranth), stem and foot, all covered by peridermal secretion, becomes recognisable. An oral constriction divides the hypostomal portion of the gastral cavity from the stomachic portion. Within the hypostomal entoderm, cells containing secretory granules differentiate. Following growth and the multiplication of tentacles, the head periderm disappears. A ring of gland cells differentiates at the hydranth's base. The positioning of cnidae in the tentacle ectoderm, penetration of the mouth opening and the multiplication of digestive gland cells enable the polyp to change from lecithotrophic to planktotrophic nutrition.

  7. Morphogenesis of the somatic musculature in Drosophila melanogaster

    PubMed Central

    Schulman, Victoria K.; Dobi, Krista C.; Baylies, Mary K.

    2015-01-01

    In Drosophila melanogaster, the somatic muscle system is first formed during embryogenesis, giving rise to the larval musculature. Later during metamorphosis, this system is destroyed and replaced by an entirely new set of muscles in the adult fly. Proper formation of the larval and adult muscles is critical for basic survival functions such as hatching and crawling (in the larva), walking and flying (in the adult), and feeding (at both larval and adult stages). Myogenesis, from mononucleated muscle precursor cells to multinucleated functional muscles, is driven by a number of cellular processes that have begun to be mechanistically defined. Once themesodermal cells destined for themyogenic lineage have been specified, individual myoblasts fuse together iteratively to form syncytial myofibers. Combining cytoplasmic contents demands a level of intracellular reorganization that, most notably, leads to redistribution of the myonuclei to maximize internuclear distance. Signaling from extending myofibers induces terminal tendon cell differentiation in the ectoderm, which results in secure muscle-tendon attachments that are critical formuscle contraction. Simultaneously, muscles become innervated and undergo sarcomerogenesis to establish the contractile apparatus that will facilitate movement. The cellular mechanisms governing these morphogenetic events share numerous parallels to mammalian development, and the basic unit of all muscle, the myofiber, is conserved from flies to mammals. Thus, studies of Drosophila myogenesis and comparisons to muscle development in other systems highlight conserved regulatory programs of biomedical relevance to general muscle biology and studies of muscle disease. PMID:25758712

  8. 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

  9. Optogenetic stimulation of Drosophila heart rate at different temperatures and Ca2+ concentrations.

    PubMed

    Zhu, Yue C; Uradu, Henry; Majeed, Zana R; Cooper, Robin L

    2016-02-01

    Optogenetics is a revolutionary technique that enables noninvasive activation of electrically excitable cells. In mammals, heart rate has traditionally been modulated with pharmacological agents or direct stimulation of cardiac tissue with electrodes. However, implanted wires have been known to cause physical damage and damage from electrical currents. Here, we describe a proof of concept to optically drive cardiac function in a model organism, Drosophila melanogaster. We expressed the light sensitive channelrhodopsin protein ChR2.XXL in larval Drosophila hearts and examined light-induced activation of cardiac tissue. After demonstrating optical stimulation of larval heart rate, the approach was tested at low temperature and low calcium levels to simulate mammalian heart transplant conditions. Optical activation of ChR2.XXL substantially increased heart rate in all conditions. We have developed a system that can be instrumental in characterizing the physiology of optogenetically controlled cardiac function with an intact heart. PMID:26834237

  10. Identification and characterization of kraken, a gene encoding a putative hydrolytic enzyme in Drosophila melanogaster.

    PubMed

    Edwin Chan, H Y; Harris, S J; O'Kane, C J

    1998-11-19

    Kraken, a novel Drosophila gene isolated from a 4-8-h-old Drosophila embryo cDNA library, shows homology to a family of serine hydrolases whose common feature is that they all catalyse breakage of substrates with a carbonyl-containing group. It is a single-copy gene with at least two introns and maps to position 21D on polytene chromosomes. kraken is a member of a conserved gene family. Messenger RNA of kraken is expressed ubiquitously in early embryogenesis. Later, it is concentrated in the foregut and the posterior midgut primordium. Towards the end of embryogenesis, expression of kraken is confined to the gastric caeca. During the third-instar larval stage, kraken is expressed at low levels in the gastric caeca and parts of the gut, and at higher levels in the fat body. We suggest a role for Kraken in detoxification and digestion during embryogenesis and larval development. PMID:9831651

  11. Klf15 Is Critical for the Development and Differentiation of Drosophila Nephrocytes

    PubMed Central

    Bodmer, Rolf; Ocorr, Karen; Paululat, Achim; Hartley, Paul S.

    2015-01-01

    Insect nephrocytes are highly endocytic scavenger cells that represent the only invertebrate model for the study of human kidney podocytes. Despite their importance, nephrocyte development is largely uncharacterised. This work tested whether the insect ortholog of mammalian Kidney Krüppel-Like Factor (Klf15), a transcription factor required for mammalian podocyte differentiation, was required for insect nephrocyte development. It was found that expression of Drosophila Klf15 (dKlf15, previously known as Bteb2) was restricted to the only two nephrocyte populations in Drosophila, the garland cells and pericardial nephrocytes. Loss of dKlf15 function led to attrition of both nephrocyte populations and sensitised larvae to the xenotoxin silver nitrate. Although pericardial nephrocytes in dKlf15 loss of function mutants were specified during embryogenesis, they failed to express the slit diaphragm gene sticks and stones and did not form slit diaphragms. Conditional silencing of dKlf15 in adults led to reduced surface expression of the endocytic receptor Amnionless and loss of in vivo scavenger function. Over-expression of dKlf15 increased nephrocyte numbers and rescued age-dependent decline in nephrocyte function. The data place dKlf15 upstream of sns and Amnionless in a nephrocyte-restricted differentiation pathway and suggest dKlf15 expression is both necessary and sufficient to sustain nephrocyte differentiation. These findings explain the physiological relevance of dKlf15 in Drosophila and imply that the role of KLF15 in human podocytes is evolutionarily conserved. PMID:26301956

  12. She’s a femme fatale: low-density larval development produces good disease vectors

    PubMed Central

    Juliano, Steven A; Ribeiro, Gabriel Sylvestre; Maciel-de-Freitas, Rafael; Castro, Márcia G; Codeço, Claudia; Lourenço-de-Oliveira, Ricardo; Lounibos, L Philip

    2014-01-01

    Two hypotheses for how conditions for larval mosquitoes affect vectorial capacity make opposite predictions about the relationship of adult size and frequency of infection with vector-borne pathogens. Competition among larvae produces small adult females. The competition-susceptibility hypothesis postulates that small females are more susceptible to infection and predicts frequency of infection should decrease with size. The competition-longevity hypothesis postulates that small females have lower longevity and lower probability of becoming competent to transmit the pathogen and thus predicts frequency of infection should increase with size. We tested these hypotheses for Aedes aegypti in Rio de Janeiro, Brazil, during a dengue outbreak. In the laboratory, longevity increases with size, then decreases at the largest sizes. For field-collected females, generalised linear mixed model comparisons showed that a model with a linear increase of frequency of dengue with size produced the best Akaike’s information criterion with a correction for small sample sizes (AICc). Consensus prediction of three competing models indicated that frequency of infection increases monotonically with female size, consistent with the competition-longevity hypothesis. Site frequency of infection was not significantly related to site mean size of females. Thus, our data indicate that uncrowded, low competition conditions for larvae produce the females that are most likely to be important vectors of dengue. More generally, ecological conditions, particularly crowding and intraspecific competition among larvae, are likely to affect vector-borne pathogen transmission in nature, in this case via effects on longevity of resulting adults. Heterogeneity among individual vectors in likelihood of infection is a generally important outcome of ecological conditions impacting vectors as larvae. PMID:25591112

  13. A core transcriptional network for early mesoderm development in Drosophila melanogaster

    PubMed Central

    Sandmann, Thomas; Girardot, Charles; Brehme, Marc; Tongprasit, Waraporn; Stolc, Viktor; Furlong, Eileen E.M.

    2007-01-01

    Embryogenesis is controlled by large gene-regulatory networks, which generate spatially and temporally refined patterns of gene expression. Here, we report the characteristics of the regulatory network orchestrating early mesodermal development in the fruitfly Drosophila, where the transcription factor Twist is both necessary and sufficient to drive development. Through the integration of chromatin immunoprecipitation followed by microarray analysis (ChIP-on-chip) experiments during discrete time periods with computational approaches, we identified >2000 Twist-bound cis-regulatory modules (CRMs) and almost 500 direct target genes. Unexpectedly, Twist regulates an almost complete cassette of genes required for cell proliferation in addition to genes essential for morophogenesis and cell migration. Twist targets almost 25% of all annotated Drosophila transcription factors, which may represent the entire set of regulators necessary for the early development of this system. By combining in vivo binding data from Twist, Mef2, Tinman, and Dorsal we have constructed an initial transcriptional network of early mesoderm development. The network topology reveals extensive combinatorial binding, feed-forward regulation, and complex logical outputs as prevalent features. In addition to binary activation and repression, we suggest that Twist binds to almost all mesodermal CRMs to provide the competence to integrate inputs from more specialized transcription factors. PMID:17322403

  14. Involvement of a putative allatostatin in regulation of juvenile hormone titer and the larval development in Leptinotarsa decemlineata (Say).

    PubMed

    Meng, Qing-Wei; Liu, Xin-Ping; Lü, Feng-Gong; Fu, Kai-Yun; Guo, Wen-Chao; Li, Guo-Qing

    2015-01-01

    Juvenile hormone III (JH III) plays primary roles in regulation of metamorphosis, reproduction and diapause in Leptinotarsa decemlineata, a notorious defoliator of potato. The neurosecretory cell-borne substance(s) negatively affects the final two steps in JH biosynthesis, catalyzed respectively by an epoxidase CYP15A1 and a juvenile hormone acid methyltransferase (JHAMT). In a few insect species other than L. decemlineata, the inhibitory substance is allatostatin (AS) neuropeptide. In this study, two putative AS genes encoding LdAS-C and LdAS-B precursors were cloned. Both LdAS-C and LdAS-B were expressed in the egg, larvae, pupae and adults, and highly expressed in the brain and the gut. Dietary introduction of double-stranded RNAs (dsRNAs) targeting LdAS-C and LdAS-B successfully knocked down respective target genes. Ingestion during 3 and 6 consecutive days of dsLdAS-C significantly increased the LdJHAMT mRNA levels by 3.8 and 9.9 fold respectively. In contrast, ingestion of dsLdAS-B only slightly increased the LdJHAMT expression level by 1.1 and 1.7 fold. Moreover, after one, two and three days' ingestion of dsLdAS-C, the relative JH levels in the hemolymph of treated larvae were 2.5, 4.2 and 1.9 fold higher than those in control beetles. Furthermore, ingestion of dsLdAS-C and dsLdAS-B significantly affected larval growth and delayed larval development. Thus, we provide a line of experimental evidence in L. decemlineata to support the concept that AS-C acts as an allatostatin and inhibit JH biosynthesis. PMID:25452193

  15. The potential of ocean acidification on suppressing larval development in the Pacific oyster Crassostrea gigas and blood cockle Arca inflata Reeve

    NASA Astrophysics Data System (ADS)

    Li, Jiaqi; Jiang, Zengjie; Zhang, Jihong; Mao, Yuze; Bian, Dapeng; Fang, Jianguang

    2014-11-01

    We evaluated the effect of pH on larval development in larval Pacific oyster ( Crassostrea gigas) and blood cockle ( Arca inflata Reeve). The larvae were reared at pH 8.2 (control), 7.9, 7.6, or 7.3 beginning 30 min or 24 h post fertilization. Exposure to lower pH during early embryonic development inhibited larval shell formation in both species. Compared with the control, larvae took longer to reach the D-veliger stage when reared under pH 7.6 and 7.3. Exposure to lower pH immediately after fertilization resulted in significantly delayed shell formation in the Pacific oyster larvae at pH 7.3 and blood cockle larvae at pH 7.6 and 7.3. However, when exposure was delayed until 24 h post fertilization, shell formation was only inhibited in blood cockle larvae reared at pH 7.3. Thus, the early embryonic stages were more sensitive to acidified conditions. Our results suggest that ocean acidification will have an adverse effect on embryonic development in bivalves. Although the effects appear subtle, they may accumulate and lead to subsequent issues during later larval development.

  16. Development of liquid larval diet with modified rearing system for Bactrocera dorsalis (Hendel) (Diptera:Tephritidae) for the application of sterile insect technique

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A liquid larval diet and its rearing system have been developed for mass rearing of Bactrocera dorsalis (Hendel) in Hawaii. Rearing facility in Institute of Food and Radiation Biology, Atomic Energy Research Establishment, Dhaka, Bangladesh, modified protein source from brewer's yeast to a combinat...

  17. GROWTH AND VARIATIONS IN LIPID CLASS AND FATTY ACID COMPOSITION DURING LARVAL DEVELOPMENT OF THE STONE CRAB, MENIPPE ADINA WILLIAMS AND FELDER, 1986

    EPA Science Inventory

    Larval development in decapod crustaceans is marked by variable growth patterns and changes in weight and biochemical composition. Larvae of the stone crab, Menippe adina, were mass-reared under laboratory conditions (28|C; 20o/ooS) from hatching to the megalopal stage. Growth in...

  18. EFFECTS OF SOLUBLE FRACTIONS OF USED LIGHT-WEIGHT LIGNOSULFONATE TYPE MUD AND HEXAVALENT CHROMIUM ON THE COMPLETE LARVAL DEVELOPMENT OF THE CRABS, 'RHITHROPANOPEUS HARRISII' AND 'CALLINECTES SAPIDUS'

    EPA Science Inventory

    The mud aqueous fractions (MAF) and suspended particulate phase (SPP) of lignosulfonate type mud were nontoxic to the complete larval development of Rhithropanopeus harrisii. Five percent MAF and SPP were not toxic to Callinectes sapidus. Differential survival of C. sapidus larva...

  19. Complete larval development of Thor amboinensis (De Man, 1888) Decapoda: Thoridae) described from laboratory-reared material
    and identified by DNA barcoding.

    PubMed

    Bartilotti, Cátia; Salabert, Joana; Santos, Antonina Dos

    2016-01-01

    Of the 12 species of Thor described until present date, only three (25%) have their complete larval development known. Present work describes the complete larval development of Thor amboinensis, based on laboratory-reared material. The spent females were identified through the analysis of the partial sequences of the mitochondrial DNA barcode, also used for the reconstruction of the phylogenetic relationships within the recently resurrected and recognized family Thoridae Kingsley, 1879. Eight zoeal stages and one decapodid complete this species larval development. In the genus Thor, the number of zoeal stages varies greatly from two (T. dobkini) to eight (T. amboinensis and T. floridanus). The larvae of T. ambionensis and T. floridanus are readily distinguished from each other by the ornamentation of the ventral margin of the carapace and the pereiopods development. The first zoeal stage of T. amboinensis described by Yang & Okuno (2004) and the one described in present study are very similar. A brief discussion on the morphological characters and on the number of zoeal stages of the genus, as well as of the previous larval descriptions is made. The phylogenetic analysis suggest cryptic speciation for geographical separated populations of T. amboinensis, paraphyly of the genus Eualus, and the reassignment of E. cranchii to a different genus. PMID:27395843

  20. GROWTH AND VARIATIONS IN LIPID CLASS AND FATTY ACID COMPOSITION DURING LARVAL DEVELOPMENT OF THE STONE CRAB, MENIPPE ADINA WILLIAMS AND FELDER, 1986.

    EPA Science Inventory

    Larval development in decapod crustaceans is marked by variable growth patterns and changes in weight and biochemical composition. Larvae of the stone crab, Menippe adina, were mass-reared under laboratory conditions (28?C; 20o/ooS) from hatching to the megalopa stage. Growth in...

  1. Specialized Cortex Glial Cells Accumulate Lipid Droplets in Drosophila melanogaster

    PubMed Central

    Kis, Viktor; Barti, Benjámin; Lippai, Mónika; Sass, Miklós

    2015-01-01

    Lipid droplets (LDs) are common organelles of the majority of eukaryotic cell types. Their biological significance has been extensively studied in mammalian liver cells and white adipose tissue. Although the central nervous system contains the highest relative amount and the largest number of different lipid species, neither the spatial nor the temporal distribution of LDs has been described. In this study, we used the brain of the fruitfly, Drosophila melanogaster, to investigate the neuroanatomy of LDs. We demonstrated that LDs are exclusively localised in glial cells but not in neurons in the larval nervous system. We showed that the brain’s LD pool, rather than being constant, changes dynamically during development and reaches its highest value at the beginning of metamorphosis. LDs are particularly enriched in cortex glial cells located close to the brain surface. These specialized superficial cortex glial cells contain the highest amount of LDs among glial cell types and encapsulate neuroblasts and their daughter cells. Superficial cortex glial cells, combined with subperineurial glial cells, express the Drosophila fatty acid binding protein (Dfabp), as we have demonstrated through light- and electron microscopic immunocytochemistry. To the best of our best knowledge this is the first study that describes LD neuroanatomy in the Drosophila larval brain. PMID:26148013

  2. Specialized Cortex Glial Cells Accumulate Lipid Droplets in Drosophila melanogaster.

    PubMed

    Kis, Viktor; Barti, Benjámin; Lippai, Mónika; Sass, Miklós

    2015-01-01

    Lipid droplets (LDs) are common organelles of the majority of eukaryotic cell types. Their biological significance has been extensively studied in mammalian liver cells and white adipose tissue. Although the central nervous system contains the highest relative amount and the largest number of different lipid species, neither the spatial nor the temporal distribution of LDs has been described. In this study, we used the brain of the fruitfly, Drosophila melanogaster, to investigate the neuroanatomy of LDs. We demonstrated that LDs are exclusively localised in glial cells but not in neurons in the larval nervous system. We showed that the brain's LD pool, rather than being constant, changes dynamically during development and reaches its highest value at the beginning of metamorphosis. LDs are particularly enriched in cortex glial cells located close to the brain surface. These specialized superficial cortex glial cells contain the highest amount of LDs among glial cell types and encapsulate neuroblasts and their daughter cells. Superficial cortex glial cells, combined with subperineurial glial cells, express the Drosophila fatty acid binding protein (Dfabp), as we have demonstrated through light- and electron microscopic immunocytochemistry. To the best of our best knowledge this is the first study that describes LD neuroanatomy in the Drosophila larval brain. PMID:26148013

  3. Dual functionality of O-GlcNAc transferase is required for Drosophila development.

    PubMed

    Mariappa, Daniel; Zheng, Xiaowei; Schimpl, Marianne; Raimi, Olawale; Ferenbach, Andrew T; Müller, H-Arno J; van Aalten, Daan M F

    2015-12-01

    Post-translational modification of intracellular proteins with O-linked N-acetylglucosamine (O-GlcNAc) catalysed by O-GlcNAc transferase (OGT) has been linked to regulation of diverse cellular functions. OGT possesses a C-terminal glycosyltransferase catalytic domain and N-terminal tetratricopeptide repeats that are implicated in protein-protein interactions. Drosophila OGT (DmOGT) is encoded by super sex combs (sxc), mutants of which are pupal lethal. However, it is not clear if this phenotype is caused by reduction of O-GlcNAcylation. Here we use a genetic approach to demonstrate that post-pupal Drosophila development can proceed with negligible OGT catalysis, while early embryonic development is OGT activity-dependent. Structural and enzymatic comparison between human OGT (hOGT) and DmOGT informed the rational design of DmOGT point mutants with a range of reduced catalytic activities. Strikingly, a severely hypomorphic OGT mutant complements sxc pupal lethality. However, the hypomorphic OGT mutant-rescued progeny do not produce F2 adults, because a set of Hox genes is de-repressed in F2 embryos, resulting in homeotic phenotypes. Thus, OGT catalytic activity is required up to late pupal stages, while further development proceeds with severely reduced OGT activity. PMID:26674417

  4. Quantitative analysis of bristle number in Drosophila mutants identifies genes involved in neural development

    NASA Technical Reports Server (NTRS)

    Norga, Koenraad K.; Gurganus, Marjorie C.; Dilda, Christy L.; Yamamoto, Akihiko; Lyman, Richard F.; Patel, Prajal H.; Rubin, Gerald M.; Hoskins, Roger A.; Mackay, Trudy F.; Bellen, Hugo J.

    2003-01-01

    BACKGROUND: The identification of the function of all genes that contribute to specific biological processes and complex traits is one of the major challenges in the postgenomic era. One approach is to employ forward genetic screens in genetically tractable model organisms. In Drosophila melanogaster, P element-mediated insertional mutagenesis is a versatile tool for the dissection of molecular pathways, and there is an ongoing effort to tag every gene with a P element insertion. However, the vast majority of P element insertion lines are viable and fertile as homozygotes and do not exhibit obvious phenotypic defects, perhaps because of the tendency for P elements to insert 5' of transcription units. Quantitative genetic analysis of subtle effects of P element mutations that have been induced in an isogenic background may be a highly efficient method for functional genome annotation. RESULTS: Here, we have tested the efficacy of this strategy by assessing the extent to which screening for quantitative effects of P elements on sensory bristle number can identify genes affecting neural development. We find that such quantitative screens uncover an unusually large number of genes that are known to function in neural development, as well as genes with yet uncharacterized effects on neural development, and novel loci. CONCLUSIONS: Our findings establish the use of quantitative trait analysis for functional genome annotation through forward genetics. Similar analyses of quantitative effects of P element insertions will facilitate our understanding of the genes affecting many other complex traits in Drosophila.

  5. Regulation of Drosophila Eye Development by the Transcription Factor Sine oculis

    PubMed Central

    Kwak, Su-Jin; Wang, Feng; Wang, Hui; Chen, Rui; Mardon, Graeme

    2014-01-01

    Homeodomain transcription factors of the Sine oculis (SIX) family direct multiple regulatory processes throughout the metazoans. Sine oculis (So) was first characterized in the fruit fly Drosophila melanogaster, where it is both necessary and sufficient for eye development, regulating cell survival, proliferation, and differentiation. Despite its key role in development, only a few direct targets of So have been described previously. In the current study, we aim to expand our knowledge of So-mediated transcriptional regulation in the developing Drosophila eye using ChIP-seq to map So binding regions throughout the genome. We find 7,566 So enriched regions (peaks), estimated to map to 5,952 genes. Using overlap between the So ChIP-seq peak set and genes that are differentially regulated in response to loss or gain of so, we identify putative direct targets of So. We find So binding enrichment in genes not previously known to be regulated by So, including genes that encode cell junction proteins and signaling pathway components. In addition, we analyze a subset of So-bound novel genes in the eye, and find eight genes that have previously uncharacterized eye phenotypes and may be novel direct targets of So. Our study presents a greatly expanded list of candidate So targets and serves as basis for future studies of So-mediated gene regulation in the eye. PMID:24586968

  6. Dual functionality of O-GlcNAc transferase is required for Drosophila development

    PubMed Central

    Mariappa, Daniel; Zheng, Xiaowei; Schimpl, Marianne; Raimi, Olawale; Ferenbach, Andrew T.; Müller, H.-Arno J.; van Aalten, Daan M. F.

    2015-01-01

    Post-translational modification of intracellular proteins with O-linked N-acetylglucosamine (O-GlcNAc) catalysed by O-GlcNAc transferase (OGT) has been linked to regulation of diverse cellular functions. OGT possesses a C-terminal glycosyltransferase catalytic domain and N-terminal tetratricopeptide repeats that are implicated in protein–protein interactions. Drosophila OGT (DmOGT) is encoded by super sex combs (sxc), mutants of which are pupal lethal. However, it is not clear if this phenotype is caused by reduction of O-GlcNAcylation. Here we use a genetic approach to demonstrate that post-pupal Drosophila development can proceed with negligible OGT catalysis, while early embryonic development is OGT activity-dependent. Structural and enzymatic comparison between human OGT (hOGT) and DmOGT informed the rational design of DmOGT point mutants with a range of reduced catalytic activities. Strikingly, a severely hypomorphic OGT mutant complements sxc pupal lethality. However, the hypomorphic OGT mutant-rescued progeny do not produce F2 adults, because a set of Hox genes is de-repressed in F2 embryos, resulting in homeotic phenotypes. Thus, OGT catalytic activity is required up to late pupal stages, while further development proceeds with severely reduced OGT activity. PMID:26674417

  7. Abbreviation of larval development and extension of brood care as key features of the evolution of freshwater Decapoda.

    PubMed

    Vogt, Günter

    2013-02-01

    The transition from marine to freshwater habitats is one of the major steps in the evolution of life. In the decapod crustaceans, four groups have colonized fresh water at different geological times since the Triassic, the freshwater shrimps, freshwater crayfish, freshwater crabs and freshwater anomurans. Some families have even colonized terrestrial habitats via the freshwater route or directly via the sea shore. Since none of these taxa has ever reinvaded its environment of origin the Decapoda appear particularly suitable to investigate life-history adaptations to fresh water. Evolutionary comparison of marine, freshwater and terrestrial decapods suggests that the reduction of egg number, abbreviation of larval development, extension of brood care and lecithotrophy of the first posthatching life stages are key adaptations to fresh water. Marine decapods usually have high numbers of small eggs and develop through a prolonged planktonic larval cycle, whereas the production of small numbers of large eggs, direct development and extended brood care until the juvenile stage is the rule in freshwater crayfish, primary freshwater crabs and aeglid anomurans. The amphidromous freshwater shrimp and freshwater crab species and all terrestrial decapods that invaded land via the sea shore have retained ocean-type planktonic development. Abbreviation of larval development and extension of brood care are interpreted as adaptations to the particularly strong variations of hydrodynamic parameters, physico-chemical factors and phytoplankton availability in freshwater habitats. These life-history changes increase fitness of the offspring and are obviously favoured by natural selection, explaining their multiple origins in fresh water. There is no evidence for their early evolution in the marine ancestors of the extant freshwater groups and a preadaptive role for the conquest of fresh water. The costs of the shift from relative r- to K-strategy in freshwater decapods are traded

  8. Transcriptomic Analysis of Differentially Expressed Genes During Larval Development of Rapana venosa by Digital Gene Expression Profiling

    PubMed Central

    Song, Hao; Yu, Zheng-Lin; Sun, Li-Na; Xue, Dong-Xiu; Zhang, Tao; Wang, Hai-Yan

    2016-01-01

    During the life cycle of shellfish, larval development, especially metamorphosis, has a vital influence on the dynamics, distribution, and recruitment of natural populations, as well as seed breeding. Rapana venosa, a carnivorous gastropod, is an important commercial shellfish in China, and is an ecological invader in the United States, Argentina, and France. However, information about the mechanism of its early development is still limited, because research in this area has long suffered from a lack of genomic resources. In this study, 15 digital gene expression (DGE) libraries from five developmental stages of R. venosa were constructed and sequenced on the IIIumina Hi-Sequation 2500 platform. Bioinformaticsanalysis identified numerous differentially and specifically expressed genes, which revealed that genes associated with growth, nervous system, digestive system, immune system, and apoptosis participate in important developmental processes. The functional analysis of differentially expressed genes was further implemented by gene ontology, and Kyoto encyclopedia of genes and genomes enrichment. DGE profiling provided a general picture of the transcriptomic activities during the early development of R. venosa, which may provide interesting hints for further study. Our data represent the first comparative transcriptomic information available for the early development of R. venosa, which is a prerequisite for a better understanding of the physiological traits controlling development. PMID:27194808

  9. Transcriptomic Analysis of Differentially Expressed Genes During Larval Development of Rapana venosa by Digital Gene Expression Profiling.

    PubMed

    Song, Hao; Yu, Zheng-Lin; Sun, Li-Na; Xue, Dong-Xiu; Zhang, Tao; Wang, Hai-Yan

    2016-01-01

    During the life cycle of shellfish, larval development, especially metamorphosis, has a vital influence on the dynamics, distribution, and recruitment of natural populations, as well as seed breeding. Rapana venosa, a carnivorous gastropod, is an important commercial shellfish in China, and is an ecological invader in the United States, Argentina, and France. However, information about the mechanism of its early development is still limited, because research in this area has long suffered from a lack of genomic resources. In this study, 15 digital gene expression (DGE) libraries from five developmental stages of R. venosa were constructed and sequenced on the IIIumina Hi-Sequation 2500 platform. Bioinformaticsanalysis identified numerous differentially and specifically expressed genes, which revealed that genes associated with growth, nervous system, digestive system, immune system, and apoptosis participate in important developmental processes. The functional analysis of differentially expressed genes was further implemented by gene ontology, and Kyoto encyclopedia of genes and genomes enrichment. DGE profiling provided a general picture of the transcriptomic activities during the early development of R. venosa, which may provide interesting hints for further study. Our data represent the first comparative transcriptomic information available for the early development of R. venosa, which is a prerequisite for a better understanding of the physiological traits controlling development. PMID:27194808

  10. Requirement for highly efficient pre-mRNA splicing during Drosophila early embryonic development

    PubMed Central

    Guilgur, Leonardo Gastón; Prudêncio, Pedro; Sobral, Daniel; Liszekova, Denisa; Rosa, André; Martinho, Rui Gonçalo

    2014-01-01

    Drosophila syncytial nuclear divisions limit transcription unit size of early zygotic genes. As mitosis inhibits not only transcription, but also pre-mRNA splicing, we reasoned that constraints on splicing were likely to exist in the early embryo, being splicing avoidance a possible explanation why most early zygotic genes are intronless. We isolated two mutant alleles for a subunit of the NTC/Prp19 complexes, which specifically impaired pre-mRNA splicing of early zygotic but not maternally encoded transcripts. We hypothesized that the requirements for pre-mRNA splicing efficiency were likely to vary during development. Ectopic maternal expression of an early zygotic pre-mRNA was sufficient to suppress its splicing defects in the mutant background. Furthermore, a small early zygotic transcript with multiple introns was poorly spliced in wild-type embryos. Our findings demonstrate for the first time the existence of a developmental pre-requisite for highly efficient splicing during Drosophila early embryonic development and suggest in highly proliferative tissues a need for coordination between cell cycle and gene architecture to ensure correct gene expression and avoid abnormally processed transcripts. DOI: http://dx.doi.org/10.7554/eLife.02181.001 PMID:24755291

  11. The Drosophila blood-brain barrier: development and function of a glial endothelium.

    PubMed

    Limmer, Stefanie; Weiler, Astrid; Volkenhoff, Anne; Babatz, Felix; Klämbt, Christian

    2014-01-01

    The efficacy of neuronal function requires a well-balanced extracellular ion homeostasis and a steady supply with nutrients and metabolites. Therefore, all organisms equipped with a complex nervous system developed a so-called blood-brain barrier, protecting it from an uncontrolled entry of solutes, metabolites or pathogens. In higher vertebrates, this diffusion barrier is established by polarized endothelial cells that form extensive tight junctions, whereas in lower vertebrates and invertebrates the blood-brain barrier is exclusively formed by glial cells. Here, we review the development and function of the glial blood-brain barrier of Drosophila melanogaster. In the Drosophila nervous system, at least seven morphologically distinct glial cell classes can be distinguished. Two of these glial classes form the blood-brain barrier. Perineurial glial cells participate in nutrient uptake and establish a first diffusion barrier. The subperineurial glial (SPG) cells form septate junctions, which block paracellular diffusion and thus seal the nervous system from the hemolymph. We summarize the molecular basis of septate junction formation and address the different transport systems expressed by the blood-brain barrier forming glial cells. PMID:25452710

  12. The role of the microbial environment in Drosophila post-embryonic development.

    PubMed

    Strigini, Maura; Leulier, François

    2016-11-01

    Development, growth and maturation of animals are under genetic and environmental control. Multicellular organisms interact throughout their lives with a variety of environment- and body-associated microorganisms. It has now been appreciated that the very conspicuous and varied microbial population associated with the food and the gastro-intestinal tract is a critical factor that can influence growth. Beyond the phenomenology, the mechanisms underlying the beneficial effects of microbes on development are being revealed from studies in Drosophila melanogaster, a particularly well suited system for a mechanistic understanding of host/microbiota interactions. Association of otherwise germ-free eggs with specific bacterial strains isolated from Drosophila gut samples can accelerate growth in larvae raised on restrictive diets. We review advances made possible by the exploitation of such simplified gnotobiotic systems in the search for the genes, molecules and physiological adaptations responsible for this effect in both host and microbes. Transposon mutagenesis and gene-trait match studies in bacteria can identify the key microbial genes and metabolites required for the beneficial effect, acetic acid being one of them. In the fly, functional genomic analysis, transcriptomics and metabolomics point to the modulation of systemic insulin and steroid hormone signalling as well as the regulation of intestinal physiology, including the enhancement of intestinal protease activity, as crucial mediators of the host's response. PMID:26827889

  13. Spen is required for pigment cell survival during pupal development in Drosophila.

    PubMed

    Querenet, Matthieu; Goubard, Valerie; Chatelain, Gilles; Davoust, Nathalie; Mollereau, Bertrand

    2015-06-15

    Apoptosis is required during development to eliminate superfluous cells and sculpt tissues; spatial and timed control of apoptosis ensures that the necessary number of cells is eliminated at a precise time in a given tissue. The elimination of supernumerary pigment or inter-ommatidial cells (IOCs) depends on cell-cell communication and is necessary for the formation of the honeycomb-like structure of the Drosophila eye. However, the mechanisms occurring during pupal development and controlling apoptosis of superfluous IOC in space and time remain unclear. Here, we found that split-ends (spen) is required for IOC survival at the time of removal of superfluous IOCs. Loss of spen function leads to abnormal removal of IOCs by apoptosis. We show that spen is required non-autonomously in cone cells for the survival of IOCs by positively regulating the Spitz/EGFR pathway. We propose that Spen is an important survival factor that ensures spatial control of the apoptotic wave that is necessary for the correct patterning and formation of the Drosophila eye. PMID:25872184

  14. The Drosophila blood-brain barrier: development and function of a glial endothelium

    PubMed Central

    Limmer, Stefanie; Weiler, Astrid; Volkenhoff, Anne; Babatz, Felix; Klämbt, Christian

    2014-01-01

    The efficacy of neuronal function requires a well-balanced extracellular ion homeostasis and a steady supply with nutrients and metabolites. Therefore, all organisms equipped with a complex nervous system developed a so-called blood-brain barrier, protecting it from an uncontrolled entry of solutes, metabolites or pathogens. In higher vertebrates, this diffusion barrier is established by polarized endothelial cells that form extensive tight junctions, whereas in lower vertebrates and invertebrates the blood-brain barrier is exclusively formed by glial cells. Here, we review the development and function of the glial blood-brain barrier of Drosophila melanogaster. In the Drosophila nervous system, at least seven morphologically distinct glial cell classes can be distinguished. Two of these glial classes form the blood-brain barrier. Perineurial glial cells participate in nutrient uptake and establish a first diffusion barrier. The subperineurial glial (SPG) cells form septate junctions, which block paracellular diffusion and thus seal the nervous system from the hemolymph. We summarize the molecular basis of septate junction formation and address the different transport systems expressed by the blood-brain barrier forming glial cells. PMID:25452710

  15. Analysis of cell identity, morphology, apoptosis and mitotic activity in a primary neural cell culture system in Drosophila

    PubMed Central

    2012-01-01

    In Drosophila, most neurogenetic research is carried out in vivo. Mammalian research demonstrates that primary cell culture techniques provide a powerful model to address cell autonomous and non-autonomous processes outside their endogenous environment. We developed a cell culture system in Drosophila using wildtype and genetically manipulated primary neural tissue for long-term observations. We assessed the molecular identity of distinct neural cell types by immunolabeling and genetically expressed fluorescent cell markers. We monitored mitotic activity of cell cultures derived from wildtype and tumorous larval brains. Our system provides a powerful approach to unveil developmental processes in the nervous system and to complement studies in vivo. PMID:22554060

  16. Same but different: Larval development and gall-inducing process of a non-pollinating fig wasp compared to that of pollinating fig-wasps

    NASA Astrophysics Data System (ADS)

    Jansen-González, Sergio; Teixeira, Simone de Padua; Kjellberg, Finn; Pereira, Rodrigo A. Santinelo

    2014-05-01

    The receptacles of fig trees (Ficus spp.) can harbor a highly diversified and complex community of chalcid wasps. Functional groups of fig wasps (e.g. gallers, cleptoparasites and parasitoids) oviposit into the fig at different developmental stages, reflecting different feeding regimes for these insect larvae. There are few direct data available on larval feeding regimes and access to resources. We studied the gall induction and larval feeding strategy of an Idarnes (group flavicollis) species, a non-pollinating fig wasp (NPFW) associated to Ficus citrifolia P. Miller in Brazil. This Idarnes species shares with the pollinator characteristics such as time of oviposition, ovipositor insertion through flower and location of the egg inside plant ovaries. Nevertheless, we show that the gall induction differs considerably from that of the pollinating species. This Idarnes species relies on the induction of nucellus cell proliferation for gall formation and as the main larval resource. This strategy enables it to develop in both pollinated and unpollinated figs. The large differences between this NPFW and other fig wasps in how ovules are galled suggest that there are different ways to be a galler. A functional analysis of NPFW community structure may require descriptions of the histological processes associated with larval development.

  17. Effects of temperature and salinity on larval survival and development in the invasive shrimp Palaemon macrodactylus (Caridea: Palaemonidae) along the reproductive season

    NASA Astrophysics Data System (ADS)

    Guadalupe Vázquez, M.; Bas, Claudia C.; Kittlein, Marcelo; Spivak, Eduardo D.

    2015-05-01

    The invasive shrimp Palaemon macrodactylus is associated mainly with brackish waters. Previous studies raised the question if tolerance to low salinities differs between larvae and adults. To answer this question, the combined effects of two temperatures (20 and 25 °C) and four salinities (5, 12, 23 and 34 psu) on survival and development of larvae that hatched at the beginning, in the midpoint and near the end of a reproductive season (denoted early, middle season and late larvae respectively) were examined. The three types of larvae were able to survive and reach juvenile phase at salinities between 12 and 34 psu and at both temperatures. At 5 psu all larvae died, but 45% molted at least once. Temperature and salinity to a lesser extent, had effects on the duration of development and on the number of larval stages in all larval types. Development was longer at the lower temperature, especially in middle season and late larvae. Most early larvae reached the juvenile phase through 5 larval stages; the number of larval stages of middle season and late larvae was higher at 20 °C and in late larvae also low salinity produced extra stages. Low salinity (12 psu) and, in early and middle season larvae, low temperature produced lighter and smaller individuals. Response of larvae to environmental factors seems to be related in part to the previous conditions (maternal effects and/or embryo development conditions). The narrower salinity tolerance of larvae compared to adults and the ability of zoea I to survive at least some days at 5 psu may be related with an export larval strategy.

  18. Combinatorial control of Drosophila eye development by Eyeless, Homothorax, and Teashirt

    PubMed Central

    Bessa, Jose; Gebelein, Brian; Pichaud, Franck; Casares, Fernando; Mann, Richard S.

    2002-01-01

    In Drosophila, the development of the compound eye depends on the movement of a morphogenetic furrow (MF) from the posterior (P) to the anterior (A) of the eye imaginal disc. We define several subdomains along the A–P axis of the eye disc that express distinct combinations of transcription factors. One subdomain, anterior to the MF, expresses two homeobox genes, eyeless (ey) and homothorax (hth), and the zinc-finger gene teashirt (tsh). We provide evidence that this combination of transcription factors may function as a complex and that it plays at least two roles in eye development: it blocks the expression of later-acting transcription factors in the eye development cascade, and it promotes cell proliferation. A key step in the transition from an immature proliferative state to a committed state in eye development is the repression of hth by the BMP-4 homolog Decapentaplegic (Dpp). PMID:12231630

  19. VARIATION IN GROWTH, LIPID CLASS AND FATTY ACID COMPOSITION OF THE MUD CRAB, RHITHROPANOPEUS HARRISII (GOULD) DURING LARVAL DEVELOPMENT FOLLOWING EXPOSURE TO AN INSECT JUVENILE HORMONE ANALOG (FENOXYCARB(R))

    EPA Science Inventory

    This study examines the effects of fenoxycarb?, an insect juvenile hormone (JH) analog, on larval growth, and lipid class and fatty acid composition in first crabs of the mud crab Rhithropanopeus harrisii reared through total larval development in nominal water concentrations fr...

  20. Effects of sister chromatid cohesion proteins on cut gene expression during wing development in Drosophila

    PubMed Central

    Dorsett, Dale; Eissenberg, Joel C.; Misulovin, Ziva; Martens, Andrew; Redding, Bethany; McKim, Kim

    2006-01-01

    Summary The cohesin protein complex is a conserved structural component of chromosomes. Cohesin binds numerous sites along interphase chromosomes and is essential for sister chromatid cohesion and DNA repair. Here, we test the idea that cohesin also regulates gene expression. This idea arose from the finding that the Drosophila Nipped-B protein, a functional homolog of the yeast Scc2 factor that loads cohesin onto chromosomes, facilitates the transcriptional activation of certain genes by enhancers located many kilobases away from their promoters. We find that cohesin binds between a remote wing margin enhancer and the promoter at the cut locus in cultured cells, and that reducing the dosage of the Smc1 cohesin subunit increases cut expression in the developing wing margin. We also find that cut expression is increased by a unique pds5 gene mutation that reduces the binding of cohesin to chromosomes. On the basis of these results, we posit that cohesin inhibits long-range activation of the Drosophila cut gene, and that Nipped-B facilitates activation by regulating cohesin-chromosome binding. Such effects of cohesin on gene expression could be responsible for many of the developmental deficits that occur in Cornelia de Lange syndrome, which is caused by mutations in the human homolog of Nipped-B. PMID:16207752

  1. mef2 activity levels differentially affect gene expression during Drosophila muscle development

    PubMed Central

    Elgar, Stuart J.; Han, Jun; Taylor, Michael V.

    2008-01-01

    Cell differentiation is controlled by key transcription factors, and a major question is how they orchestrate cell-type-specific genetic programs. Muscle differentiation is a well studied paradigm in which the conserved Mef2 transcription factor plays a pivotal role. Recent genomic studies have identified a large number of mef2-regulated target genes with distinct temporal expression profiles during Drosophila myogenesis. However, the question remains as to how a single transcription factor can control such diverse patterns of gene expression. In this study we used a strategy combining genomics and developmental genetics to address this issue in vivo during Drosophila muscle development. We found that groups of mef2-regulated genes respond differently to changes in mef2 activity levels: some require higher levels for their expression than others. Furthermore, this differential requirement correlates with when the gene is first expressed during the muscle differentiation program. Genes that require higher levels are activated later. These results implicate mef2 in the temporal regulation of muscle gene expression, and, consistent with this, we show that changes in mef2 activity levels can alter the start of gene expression in a predictable manner. Together these results indicate that Mef2 is not an all-or-none regulator; rather, its action is more subtle, and levels of its activity are important in the differential expression of muscle genes. This suggests a route by which mef2 can orchestrate the muscle differentiation program and contribute to the stringent regulation of gene expression during myogenesis. PMID:18198273

  2. Drosophila DOCK family protein sponge regulates the JNK pathway during thorax development.

    PubMed

    Morishita, Kazushige; Ozasa, Fumito; Eguchi, Koichi; Yoshioka, Yasuhide; Yoshida, Hideki; Hiai, Hiroshi; Yamaguchi, Masamitsu

    2014-01-01

    The dedicator of cytokinesis (DOCK) family proteins that are conserved in a wide variety of species are known as DOCK1-DOCK11 in mammals. The Sponge (Spg) is a Drosophila counterpart to the mammalian DOCK3. Specific knockdown of spg by pannir-GAL4 or apterous-GAL4 driver in wing discs induced split thorax phenotype in adults. Reduction of the Drosophila c-Jun N-terminal kinase (JNK), basket (bsk) gene dose enhanced the spg knockdown-induced phenotype. Conversely, overexpression of bsk suppressed the split thorax phenotype. Monitoring JNK activity in the wing imaginal discs by immunostaining with anti-phosphorylated JNK (anti-pJNK) antibody together with examination of lacZ expression in a puckered-lacZ enhancer trap line revealed the strong reduction of the JNK activity in the spg knockdown clones. This was further confirmed by Western immunoblot analysis of extracts from wing discs of spg knockdown fly with anti-pJNK antibody. Furthermore, the Duolink in situ Proximity Ligation Assay method detected interaction signals between Spg and Rac1 in the wing discs. Taken together, these results indicate Spg positively regulates JNK pathway that is required for thorax development and the regulation is mediated by interaction with Rac1. PMID:25311449

  3. Effete, an E2 ubiquitin-conjugating enzyme with multiple roles in Drosophila development and chromatin organization

    PubMed Central

    Cipressa, Francesca; Cenci, Giovanni

    2013-01-01

    The Drosophila effete gene encodes an extremely conserved class I E2 ubiquitin-conjugating enzyme. Growing evidence indicates that Eff is involved in many cellular processes including eye development, maintenance of female germline stem cells, and regulation of apoptosis. Eff is also a major component of Drosophila chromatin and it is particularly enriched in chromatin with repressive properties. In addition, Eff is required for telomere protection and to prevent telomere fusion. Consistent with its multiple roles in chromatin maintenance, Eff is also one of the rare factors that modulate both telomere-induced and heterochromatin-induced position effect variegation. PMID:24088712

  4. ParaHox gene expression in larval and postlarval development of the polychaete Nereis virens (Annelida, Lophotrochozoa)

    PubMed Central

    Kulakova, Milana A; Cook, Charles E; Andreeva, Tatiana F

    2008-01-01

    Background Transcription factors that encode ANTP-class homeobox genes play crucial roles in determining the body plan organization and specification of different organs and tissues in bilaterian animals. The three-gene ParaHox family descends from an ancestral gene cluster that existed before the evolution of the Bilateria. All three ParaHox genes are reported from deuterostomes and lophotrochozoans, but not to date from any ecdysozoan taxa, and there is evidence that the ParaHox genes, like the related Hox genes, were ancestrally a single chromosomal cluster. However, unlike the Hox genes, there is as yet no strong evidence that the ParaHox genes are expressed in spatial and temporal order during embryogenesis. Results We isolated fragments of the three Nereis virens ParaHox genes, then used these as probes for whole-mount in situ hybridization in larval and postlarval worms. In Nereis virens the ParaHox genes participate in antero-posterior patterning of ectodermal and endodermal regions of the digestive tract and are expressed in some cells in the segment ganglia. The expression of these genes occurs in larval development in accordance with the position of these cells along the main body axis and in postlarval development in accordance with the position of cells in ganglia along the antero-posterior axis of each segment. In none of these tissues does expression of the three ParaHox genes follow the rule of temporal collinearity. Conclusion In Nereis virens the ParaHox genes are expressed during antero-posterior patterning of the digestive system (ectodermal foregut and hindgut, and endodermal midgut) of Nereis virens. These genes are also expressed during axial specification of ventral neuroectodermal cell domains, where the expression domains of each gene are re-iterated in each neuromere except for the first parapodial segment. These expression domains are probably predetermined and may be directed on the antero-posterior axis by the Hox genes, whose

  5. Effect of flavan-3-ols on in vitro egg hatching, larval development and viability of infective larvae of Trichostrongylus colubriformis.

    PubMed

    Molan, A L; Meagher, L P; Spencer, P A; Sivakumaran, S

    2003-12-01

    The effects of flavan-3-ols (the monomer units of condensed tannins (CT)) and their galloyl derivatives on the viability of eggs, the development of first stage (L1) larvae, and the viability of the infective larvae of Trichostrongylus colubriformis were investigated under in vitro conditions. Each of the flavan-3-ol gallates showed some inhibition of egg hatching at 100 microg/ml, and 100% inhibition at 1000 microg/ml, with epigallocatechin gallate being the most effective in the egg hatch (EH) assay. In contrast, none of the flavan-3-ols were able to completely inhibit egg hatching. The flavan-3-ols and galloyl derivatives dose-dependently inhibited the development of infective larvae as assessed by the larval development (LD) assay. A larval migration inhibition (LMI) assay was used to assess the effect of flavan-3-ols and their galloyl derivatives on the motility of the infective third-stage (L3) larvae of T. colubriformis. In general, the flavan-3-ol gallates were more effective than the flavan-3-ols at immobilising the infective larvae as evidenced by their ability to inhibit more (P<0.05-0.01) larvae from passing through the LMI sieves. At 500 microg/ml, epigallocatechin gallate inhibited significantly more (P<0.1) larvae from passing through the sieves than did catechin gallate, epicatechin gallate, or gallocatechin gallate. Comparisons were made between the flavan-3-ols and their galloyl derivatives with the in vitro effects of CT extracts from several forage legumes, which have exhibited effects on parasites in vivo. The forage legumes tested at 200-500 microg/ml reduced the proportion of eggs that hatch, with comparable results to those obtained using the flavan-3-ols. The activities may be influenced by the prodelphinidin: procyanidin (PD:PC) ratios: CT extracts from Lotus pendunculatus and sainfoin have PD:PC ratios of 70:30 and 77:23, respectively, whereas the less active CT extract from Lotus corniculatus has a PD:PC ratio of 27:73. The active CT

  6. In vitro development of Strongylus edentatus to the fourth larval stage with notes on Strongylus vulgaris and Strongylus equinus.

    PubMed

    Farrar, R G; Klei, T R

    1985-08-01

    Strongylus edentatus was successfully cultured in vitro to the fourth larval stage (L4). Some growth continued for periods of 40-50 days at which time reductions in viability were observed in some of the culture systems tested. Various combinations of media, sera, buffers and organ explant cultures were tested. All cultures were incubated at 37 C in an atmosphere of 95% air and 5% CO2. Larvae underwent growth and differentiation to the L4 in all medium-serum combinations with and without organ explant cultures. Development and growth did occur but viability was reduced to insignificant levels in media without serum or cells. Optimal growth, differentiation, and longevity were observed in bicarbonate buffered RPMI-1640 containing 10% fetal calf serum and gerbil (Meriones unguiculatus) cecum explant cultures. Observations indicated that Strongylus vulgaris and Strongylus equinus also developed to the L4 stage using similar techniques. However, viability of S. vulgaris L4 was markedly limited. Specific morphological changes marked phases of development of S. edentatus, categorized as early, middle and late third stage, third molt and early fourth stage. Strongylus equinus appeared to follow the same developmental pattern in vitro as S. edentatus. Distinct differences in morphological features during differentiation were observed between S. edentatus and S. vulgaris. PMID:4032151

  7. The morphogenic features of otoconia during larval development of Cynops pyrrhogaster, the Japanese red-bellied newt

    NASA Technical Reports Server (NTRS)

    Steyger, P. S.; Wiederhold, M. L.; Batten, J.

    1995-01-01

    Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. Mammalian otoconia are barrel-shaped with triplanar facets at each end. Reptilian otoconia are commonly prismatic or fusiform in shape. Amphibians have all three otoconial morphologies, barrel-shaped otoconia within the utricle, with prismatic and fusiform otoconia in the saccule. Scanning electron microscopy revealed a sequential appearance of all three otoconial morphologies during larval development of the newt, Cynops pyrrhogaster. The first otoconia appear within a single, developing otolith, and some resemble adult barrel-shaped otoconia. As the larvae hatch, around stages 39-42, the single otolith divides into two anatomically separate regions, the utricle and saccule, and both contain otoconia similar to those seen in the single otolith. Throughout development, these otoconia may have variable morphologies, with serrated surfaces, or circumferential striations with either separated facets or adjacent facets in the triplanar end-regions. Small fusiform otoconia occur later, at stage 51, and only in the saccule. Prismatic otoconia appear later still, at stage 55, and again only in the saccule. Thus, although prismatic otoconia are the most numerous in adult newts, it is the last vestibular otoconial morphology to be expressed.

  8. Heterochrony in mandible development of larval shrimp (Decapoda: Caridea)--a comparative morphological SEM study of two carideans.

    PubMed

    Batel, Annika; Melzer, Roland R; Anger, Klaus; Geiselbrecht, Hannes

    2014-11-01

    Mandible development in the larval stages I-V of two palaemonid shrimp species, Palaemon elegans and Macrobrachium amazonicum, was analyzed using scanning electron microscopy, light microscopy, and confocal laser scanning microscopy. In contrast to the zoea I of P. elegans, first-stage larvae of M. amazonicum are nonfeeding. At hatching, the morphology of the mandibles is fully expressed in P. elegans, while it appears underdeveloped in M. amazonicum, presenting only small precursors of typical caridean features. In successive zoeal stages, both species show similar developmental changes, but the mandibular characters of the larvae in M. amazonicum were delayed compared to the equivalent stages in P. elegans, especially in the development of submarginal setae and mandible size. In conclusion, our results indicate heterochrony (postdisplacement) of mandible development in M. amazonicum compared to that in P. elegans, which is related to initial lack of mandible functionality or planktivorous feeding at hatching, respectively. This conclusion is supported by comparison with other palaemonid zoeae exhibiting different feeding modes. Our data suggest that an evolutionary ground pattern of mandible morphology is present even in species with nonfeeding first-stage larvae. PMID:24888760

  9. Eukaryotic translation initiation factor 5B activity regulates larval growth rate and germline development in Caenorhabditis elegans.

    PubMed

    Yu, Xiang; Vought, Valarie E; Conradt, Barbara; Maine, Eleanor M

    2006-09-01

    In C. elegans, a population of proliferating germ cells is maintained via GLP-1/Notch signaling; in the absence of GLP-1 signaling, germ cells prematurely enter meiosis and differentiate. We previously identified ego (enhancer of glp-1) genes that promote germline proliferation and interact genetically with the GLP-1 signaling pathway. Here, we report that iffb-1 (initiation factor five B) is an ego gene. iffb-1 encodes the sole C. elegans isoform of eukaryotic translation initiation factor 5B, a protein essential for translation. We have used RNA interference and a deletion mutation to determine the developmental consequences of reduced iffb-1 activity. Our data indicate that maternal iffb-1 gene expression is sufficient for embryogenesis, and zygotic iffb-1 expression is required for development beyond late L1/early L2 stage. Partial reduction in iffb-1 expression delays larval development and can severely disrupt proliferation and differentiation of germ cells. We hypothesize that germline development is particularly sensitive to iffb-1 expression level. PMID:16937415

  10. Effects of a fungicide formulation on embryo-larval development, metamorphosis, and gonadogenesis of the South American toad Rhinella arenarum.

    PubMed

    Svartz, Gabriela; Meijide, Fernando; Pérez Coll, Cristina

    2016-07-01

    Sublethal toxicity of the formulated fungicide Maxim(®) XL on embryonic, larval and juvenile development of Rhinella arenarum was evaluated by means of standardized bioassays. Maxim(®) XL, one of the most used fungicides in Argentina, is based on a mixture of two active ingredients: Fludioxonil and Metalaxyl-M. Maxim(®) XL exposure induced severe sublethal effects on the embryos, expressed as general underdevelopment, axial flexures, microcephaly, cellular dissociation, abnormal pigmentation, underdeveloped gills, marked edema and wavy tail. As the embryo development advanced, alterations in behavior as spasmodic contractions, general weakness and inanition were observed. Maxim(®) XL did not affect neither the time required to complete metamorphosis nor sex proportions, but gonadal development and differentiation were impaired. Gross gonadal analysis revealed a significant proportion of exposed individuals with underdevelopment of one or both gonads. Histological analysis confirmed that 18% and 10% of the individuals exposed to 0.25 and 2mg/L Maxim(®) XL, respectively, exhibited undifferentiated gonads characterized by a reduced number (or absence) of germ cells. Taking into account the risk evaluation performed by means of Hazard Quotients, this fungicide could be a threat to R. arenarum populations under chronic exposure. This study represents the first evidence of toxic effects exerted by Maxim(®) XL on amphibians. Finally, our findings highlight the properties of this fungicide that might jeopardize non-target living species exposed to it in agricultural environments. PMID:27214195

  11. Sumoylation Modulates the Activity of Spalt-like Proteins during Wing Development in Drosophila*

    PubMed Central

    Sánchez, Jonatan; Talamillo, Ana; Lopitz-Otsoa, Fernando; Pérez, Coralia; Hjerpe, Roland; Sutherland, James D.; Herboso, Leire; Rodríguez, Manuel S.; Barrio, Rosa

    2010-01-01

    The Spalt-like family of zinc finger transcription factors is conserved throughout evolution and is involved in fundamental processes during development and during embryonic stem cell maintenance. Although human SALL1 is modified by SUMO-1 in vitro, it is not known whether this post-translational modification plays a role in regulating the activity of this family of transcription factors. Here, we show that the Drosophila Spalt transcription factors are modified by sumoylation. This modification influences their nuclear localization and capacity to induce vein formation through the regulation of target genes during wing development. Furthermore, spalt genes interact genetically with the sumoylation machinery to repress vein formation in intervein regions and to attain the wing final size. Our results suggest a new level of regulation of Sall activity in vivo during animal development through post-translational modification by sumoylation. The evolutionary conservation of this family of transcription factors suggests a functional role for sumoylation in vertebrate Sall members. PMID:20562097

  12. Effects of juvenile hormone (JH) analog insecticides on larval development and JH esterase activity in two spodopterans.

    PubMed

    El-Sheikh, El-Sayed A; Kamita, Shizuo G; Hammock, Bruce D

    2016-03-01

    Juvenile hormone analog (JHA) insecticides are biological and structural mimics of JH, a key insect developmental hormone. Toxic and anti-developmental effects of the JHA insecticides methoprene, fenoxycarb, and pyriproxyfen were investigated on the larval and pupal stages of Spodoptera littoralis and Spodoptera frugiperda. Bioassays showed that fenoxycarb has the highest toxicity and fastest speed of kill in 2nd instar S. littoralis. All three JHAs affected the development of 6th instar (i.e., final instar) and pupal S. frugiperda. JH esterase (JHE) is a critical enzyme that helps to regulate JH levels during insect development. JHE activity in the last instar S. littoralis and S. frugiperda was 11 and 23 nmol min(-1) ml(-1) hemolymph, respectively. Methoprene and pyriproxyfen showed poor inhibition of JHE activity from these insects, whereas fenoxycarb showed stronger inhibition. The inhibitory activity of fenoxycarb, however, was more than 1000-fold lower than that of OTFP, a highly potent inhibitor of JHEs. Surprisingly, topical application of methoprene, fenoxycarb or pyriproxyfen on 6th instars of S. littoralis and S. frugiperda prevented the dramatic reduction in JHE activity that was found in control insects. Our findings suggest that JHAs may function as JH agonists that play a disruptive role or a hormonal replacement role in S. littoralis and S. frugiperda. PMID:26969437

  13. Impacts of regular and random noise on the behaviour, growth and development of larval Atlantic cod (Gadus morhua)

    PubMed Central

    Nedelec, Sophie L.; Simpson, Stephen D.; Morley, Erica L.; Nedelec, Brendan; Radford, Andrew N.

    2015-01-01

    Anthropogenic noise impacts behaviour and physiology in many species, but responses could change with repeat exposures. As repeat exposures can vary in regularity, identifying regimes with less impact is important for regulation. We use a 16-day split-brood experiment to compare effects of regular and random acoustic noise (playbacks of recordings of ships), relative to ambient-noise controls, on behaviour, growth and development of larval Atlantic cod (Gadus morhua). Short-term noise caused startle responses in newly hatched fish, irrespective of rearing noise. Two days of both regular and random noise regimes reduced growth, while regular noise led to faster yolk sac use. After 16 days, growth in all three sound treatments converged, although fish exposed to regular noise had lower body width–length ratios. Larvae with lower body width–length ratios were easier to catch in a predator-avoidance experiment. Our results demonstrate that the timing of acoustic disturbances can impact survival-related measures during development. Much current work focuses on sound levels, but future studies should consider the role of noise regularity and its importance for noise management and mitigation measures. PMID:26468248

  14. Impacts of regular and random noise on the behaviour, growth and development of larval Atlantic cod (Gadus morhua).

    PubMed

    Nedelec, Sophie L; Simpson, Stephen D; Morley, Erica L; Nedelec, Brendan; Radford, Andrew N

    2015-10-22

    Anthropogenic noise impacts behaviour and physiology in many species, but responses could change with repeat exposures. As repeat exposures can vary in regularity, identifying regimes with less impact is important for regulation. We use a 16-day split-brood experiment to compare effects of regular and random acoustic noise (playbacks of recordings of ships), relative to ambient-noise controls, on behaviour, growth and development of larval Atlantic cod (Gadus morhua). Short-term noise caused startle responses in newly hatched fish, irrespective of rearing noise. Two days of both regular and random noise regimes reduced growth, while regular noise led to faster yolk sac use. After 16 days, growth in all three sound treatments converged, although fish exposed to regular noise had lower body width-length ratios. Larvae with lower body width-length ratios were easier to catch in a predator-avoidance experiment. Our results demonstrate that the timing of acoustic disturbances can impact survival-related measures during development. Much current work focuses on sound levels, but future studies should consider the role of noise regularity and its importance for noise management and mitigation measures. PMID:26468248

  15. Starvation stress during larval development facilitates an adaptive response in adult worker honey bees (Apis mellifera L.).

    PubMed

    Wang, Ying; Kaftanoglu, Osman; Brent, Colin S; Page, Robert E; Amdam, Gro V

    2016-04-01

    Most organisms are constantly faced with environmental changes and stressors. In diverse organisms, there is an anticipatory mechanism during development that can program adult phenotypes. The adult phenotype would be adapted to the predicted environment that occurred during organism maturation. However, whether this anticipatory mechanism is present in eusocial species is questionable because eusocial organisms are largely shielded from exogenous conditions by their stable nest environment. In this study, we tested whether food deprivation during development of the honey bee (Apis mellifera), a eusocial insect model, can shift adult phenotypes to better cope with nutritional stress. After subjecting fifth instar worker larvae to short-term starvation, we measured nutrition-related morphology, starvation resistance, physiology, endocrinology and behavior in the adults. We found that the larval starvation caused adult honey bees to become more resilient toward starvation. Moreover, the adult bees were characterized by reduced ovary size, elevated glycogen stores and juvenile hormone (JH) titers, and decreased sugar sensitivity. These changes, in general, can help adult insects survive and reproduce in food-poor environments. Overall, we found for the first time support for an anticipatory mechanism in a eusocial species, the honey bee. Our results suggest that this mechanism may play a role in honey bee queen-worker differentiation and worker division of labor, both of which are related to the responses to nutritional stress. PMID:27030775

  16. The Drosophila Wilms׳ Tumor 1-Associating Protein (WTAP) homolog is required for eye development.

    PubMed

    Anderson, Abigail M; Weasner, Brandon P; Weasner, Bonnie M; Kumar, Justin P

    2014-06-15

    Sine Oculis (So), the founding member of the SIX family of homeobox transcription factors, binds to sequence specific DNA elements and regulates transcription of downstream target genes. It does so, in part, through the formation of distinct biochemical complexes with Eyes Absent (Eya) and Groucho (Gro). While these complexes play significant roles during development, they do not account for all So-dependent activities in Drosophila. It is thought that additional So-containing complexes make important contributions as well. This contention is supported by the identification of nearly two-dozen additional proteins that complex with So. However, very little is known about the roles that these additional complexes play in development. In this report we have used yeast two-hybrid screens and co-immunoprecipitation assays from Kc167 cells to identify a biochemical complex consisting of So and Fl(2)d, the Drosophila homolog of human Wilms׳ Tumor 1-Associating Protein (WTAP). We show that Fl(2)d protein is distributed throughout the entire eye-antennal imaginal disc and that loss-of-function mutations lead to perturbations in retinal development. The eye defects are manifested behind the morphogenetic furrow and result in part from increased levels of the pan-neuronal RNA binding protein Embryonic Lethal Abnormal Vision (Elav) and the RUNX class transcription factor Lozenge (Lz). We also provide evidence that So and Fl(2)d interact genetically in the developing eye. Wilms׳ tumor-1 (WT1), a binding partner of WTAP, is required for normal eye formation in mammals and loss-of-function mutations are associated with some versions of retinoblastoma. In contrast, WTAP and its homologs have not been implicated in eye development. To our knowledge, the results presented in this report are the first description of a role for WTAP in the retina of any seeing animal. PMID:24690230

  17. A Cell Marker System and Mosaic Patterns during Early Embryonic Development in Drosophila melanogaster

    PubMed Central

    Zusman, Susan B.; Wieschaus, Eric

    1987-01-01

    An embryonic cell marker system has been developed in Drosophila melanogaster that has enabled us to identify the genotype of cells as early as the cellular blastoderm stage of development. This system allows unambiguous detection of embryos homozygous for most X-linked lethal mutations at stages prior to when their first defects become obvious. By examining gynandromorphs at this stage, we have observed that the number of nuclei per unit area in male regions is about half that in female regions. An examination of early cleavage stage embryos whose DNA has been stained with Hoechst 33258 and whose actin has been stained with phalloidin suggests that this difference is due to a cell cycle delay in cells losing the ring-X. These experiments also demonstrate the existence of a mechanism which controls the timing of nuclear divisions in cycle 10–14 embryos. PMID:3108074

  18. Senseless, a Zn finger transcription factor, is necessary and sufficient for sensory organ development in Drosophila

    NASA Technical Reports Server (NTRS)

    Nolo, R.; Abbott, L. A.; Bellen, H. J.

    2000-01-01

    The senseless (sens) gene is required for proper development of most cell types of the embryonic and adult peripheral nervous system (PNS) of Drosophila. Sens is a nuclear protein with four Zn fingers that is expressed and required in the sensory organ precursors (SOP) for proper proneural gene expression. Ectopic expression of Sens in many ectodermal cells causes induction of PNS external sensory organ formation and is able to recreate an ectopic proneural field. Hence, sens is both necessary and sufficient for PNS development. Our data indicate that proneural genes activate sens expression. Sens is then in turn required to further activate and maintain proneural gene expression. This feedback mechanism is essential for selective enhancement and maintenance of proneural gene expression in the SOPs.

  19. Interaction between Ataxin-2 Binding Protein 1 and Cubitus-interruptus during wing development in Drosophila.

    PubMed

    Usha, N; Shashidhara, L S

    2010-05-15

    Animal growth and development is dependent on reiterative use of key signaling pathways such as Hedgehog (Hh) pathway. It is widely believed that Cubitus-interruptus (Ci) mediates all functions of Hh pathway. Here we report that CG32062, the Drosophila homologue of Ataxin-2 Binding Protein 1 (dA2BP1), functions as a cofactor of Ci to specify intervein region between L3 and L4 veins of the adult wing. Specifically, Ci-mediated transactivation of knot/collier (kn) in this region of the developing wing imaginal disc is dependent on dA2BP1 function. Protein interaction studies and chromatin-immunoprecipiation experiments suggest that Ci helps dA2BP1 to bind kn promoter, which in turn may help Ci to activate kn expression. These results suggest a mechanism by which Ci may activate targets such as kn, which do not have classical Ci/Gli-binding sites. PMID:20226779

  20. mus304 encodes a novel DNA damage checkpoint protein required during Drosophila development

    PubMed Central

    Brodsky, Michael H.; Sekelsky, Jeff J.; Tsang, Garson; Hawley, R. Scott; Rubin, Gerald M.

    2000-01-01

    Checkpoints block cell cycle progression in eukaryotic cells exposed to DNA damaging agents. We show that several Drosophila homologs of checkpoint genes, mei-41, grapes, and 14-3-3ε, regulate a DNA damage checkpoint in the developing eye. We have used this assay to show that the mutagen-sensitive gene mus304 is also required for this checkpoint. mus304 encodes a novel coiled-coil domain protein, which is targeted to the cytoplasm. Similar to mei-41, mus304 is required for chromosome break repair and for genomic stability. mus304 animals also exhibit three developmental defects, abnormal bristle morphology, decreased meiotic recombination, and arrested embryonic development. We suggest that these phenotypes reflect distinct developmental consequences of a single underlying checkpoint defect. Similar mechanisms may account for the puzzling array of symptoms observed in humans with mutations in the ATM tumor suppressor gene. PMID:10733527

  1. Comparison of somatic reversions between the ivory allele and transposon-caused mutant alleles at the white locus of Drosophila melanogaster after larval treatment with X rays and ethyl methanesulfonate

    SciTech Connect

    Ryo, H.; Yoo, M.A.; Fujikawa, K.; Kondo, S.

    1985-07-01

    Somatic reversion of strains with the ivory (wi) allele, a mutation associated with a tandem duplication of a DNA sequence at the white locus, increased with the age of larvae at the time of X-irradiation as expected from the increase in the number of target cells. In contrast, two independently isolated strains with unstable w+ loci associated with insertion of transposable elements showed higher reversion frequencies after treatment with X rays or ethyl methanesulfonate (EMS) at early larval stages than at late stages. Nevertheless, both the wi strain and the two unstable w+ strains reverted at nearly equal rates after treatment with X rays or EMS at early larval stages. Possible similarity in hot spot structure for the high reversibility of the two types of mutations is discussed in relation to production of presumed mutator-type cofactors specific to the transposon-caused mutations at early larval stages.

  2. Effects of proteinase inhibitor from Adenanthera pavonina seeds on short- and long term larval development of Aedes aegypti.

    PubMed

    Sasaki, Daniele Yumi; Jacobowski, Ana Cristina; de Souza, Antônio Pancrácio; Cardoso, Marlon Henrique; Franco, Octávio Luiz; Macedo, Maria Lígia Rodrigues

    2015-05-01

    Currently, one of the major global public health concerns is related to the transmission of dengue/yellow fever virus by the vector Aedes aegypti. The most abundant digestive enzymes in Ae. aegypti midgut larvae are trypsin and chymotrypsin. Since protease inhibitors have the capacity to bind to and inhibit the action of insect digestive proteinases, we investigated the short- and long-term effects of Adenanthera pavonina seed proteinase inhibitor (ApTI) on Ae. aegypti larvae, as well as a possible mechanism of adaptation. ApTI had a significant effect on Ae. aegypti larvae exposed to a non-lethal concentration of ApTI during short- and long-duration assays, decreasing survival, weight and proteinase activities of midgut extracts of larvae. The zymographic profile of ApTI demonstrated seven bands; three bands apparently have trypsin-like activity. Moreover, the peritrophic membrane was not disrupted. The enzymes of ApTI-fed larvae were found to be sensitive to ApTI and to have a normal feedback mechanism; also, the larval digestive enzymes were not able to degrade the inhibitor. In addition, ApTI delayed larval development time. Histological studies demonstrated a degeneration of the microvilli of the posterior midgut region epithelium cells, hypertrophy of the gastric caeca cells and an augmented ectoperitrophic space in larvae. Moreover, Ae. aegypti larvae were incapable of overcoming the negative effects of ApTI, indicating that this inhibitor might be used as a promising agent against Ae. aegypti. In addition, molecular modeling and molecular docking studies were also performed in order to construct three-dimensional theoretical models for ApTI, trypsin and chymotrypsin from Ae. aegypti, as well as to predict the possible interactions and affinity values for the complexes ApTI/trypsin and ApTI/chymotrypsin. In this context, this study broadens the base of our understanding about the modes of action of proteinase inhibitors in insects, as well as the way insects

  3. Genome-wide analysis of mRNA decay patterns during early Drosophila development

    PubMed Central

    2010-01-01

    Background The modulation of mRNA levels across tissues and time is key for the establishment and operation of the developmental programs that transform the fertilized egg into a fully formed embryo. Although the developmental mechanisms leading to differential mRNA synthesis are heavily investigated, comparatively little attention is given to the processes of mRNA degradation and how these relate to the molecular programs controlling development. Results Here we combine timed collection of Drosophila embryos and unfertilized eggs with genome-wide microarray technology to determine the degradation patterns of all mRNAs present during early fruit fly development. Our work studies the kinetics of mRNA decay, the contributions of maternally and zygotically encoded factors to mRNA degradation, and the ways in which mRNA decay profiles relate to gene function, mRNA localization patterns, translation rates and protein turnover. We also detect cis-regulatory sequences enriched in transcripts with common degradation patterns and propose several proteins and microRNAs as developmental regulators of mRNA decay during early fruit fly development. Finally, we experimentally validate the effects of a subset of cis-regulatory sequences and trans-regulators in vivo. Conclusions Our work advances the current understanding of the processes controlling mRNA degradation during early Drosophila development, taking us one step closer to the understanding of mRNA decay processes in all animals. Our data also provide a valuable resource for further experimental and computational studies investigating the process of mRNA decay. PMID:20858238

  4. Morphological Markers for Cryopreservation in the Embryonic Development of Drosophila suzukii (Diptera: Drosophilidae).

    PubMed

    Landi, Silvia; Gargani, Elisabetta; Paoli, Francesco; Simoni, Sauro; Roversi, Pio Federico

    2015-08-01

    Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is an invasive pest recently reported in Europe whose spread into new areas has caused severe economic damage to many agricultural crops. There are serious concerns about the currently available chemical insecticides because of their low efficacy in controlling the species and their environmental impact; so, several studies have focused on environmentally safe strategies. The sterile insect technique (SIT), which requires colony maintenance in laboratory and production of large numbers of live animals, can be utilized in pest management programs and could be integrated with other control strategies if the potential risks associated with the rearing and maintenance of the insect line under laboratory conditions are given sufficient attention. In this regard, the ability to cryobiologically preserve such stocks would be of substantial value. Important prerequisites for long-term cryopreservation are determination of the embryonic stages, identification of specific embryonic stages, and knowledge of development time. This paper describes the main visible markers for the different stages of embryonic development and determines the timing of development at 25°C. D. suzukii embryogenesis lasts 23-25 h at 25°C and can be divided into 17 stages defined by specific morphological markers. The point at which 50% of embryos are at Stage 14 and 50% are at Stage 15, the most tolerant stages for cryopreservation treatment, as ascertained for Drosophila melanogaster Meigen in prior studies, is reached in 14-15 h. The efficiency of this procedure might be impaired by the retention of eggs in the oviducts, making it impossible to determine the stage of embryonic development for ∼25% of laid eggs. PMID:26470330

  5. Class I Myosins Have Overlapping and Specialized Functions in Left-Right Asymmetric Development in Drosophila

    PubMed Central

    Okumura, Takashi; Sasamura, Takeshi; Inatomi, Momoko; Hozumi, Shunya; Nakamura, Mitsutoshi; Hatori, Ryo; Taniguchi, Kiichiro; Nakazawa, Naotaka; Suzuki, Emiko; Maeda, Reo; Yamakawa, Tomoko; Matsuno, Kenji

    2015-01-01

    The class I myosin genes are conserved in diverse organisms, and their gene products are involved in actin dynamics, endocytosis, and signal transduction. Drosophila melanogaster has three class I myosin genes, Myosin 31DF (Myo31DF), Myosin 61F (Myo61F), and Myosin 95E (Myo95E). Myo31DF, Myo61F, and Myo95E belong to the Myosin ID, Myosin IC, and Myosin IB families, respectively. Previous loss-of-function analyses of Myo31DF and Myo61F revealed important roles in left–right (LR) asymmetric development and enterocyte maintenance, respectively. However, it was difficult to elucidate their roles in vivo, because of potential redundant activities. Here we generated class I myosin double and triple mutants to address this issue. We found that the triple mutant was viable and fertile, indicating that all three class I myosins were dispensable for survival. A loss-of-function analysis revealed further that Myo31DF and Myo61F, but not Myo95E, had redundant functions in promoting the dextral LR asymmetric development of the male genitalia. Myo61F overexpression is known to antagonize the dextral activity of Myo31DF in various Drosophila organs. Thus, the LR-reversing activity of overexpressed Myo61F may not reflect its physiological function. The endogenous activity of Myo61F in promoting dextral LR asymmetric development was observed in the male genitalia, but not the embryonic gut, another LR asymmetric organ. Thus, Myo61F and Myo31DF, but not Myo95E, play tissue-specific, redundant roles in LR asymmetric development. Our studies also revealed differential colocalization of the class I myosins with filamentous (F)-actin in the brush border of intestinal enterocytes. PMID:25659376

  6. Myosin VIIA, Important for Human Auditory Function, Is Necessary for Drosophila Auditory Organ Development

    PubMed Central

    Todi, Sokol V.; Sivan-Loukianova, Elena; Jacobs, Julie S.; Kiehart, Daniel P.; Eberl, Daniel F.

    2008-01-01

    Background Myosin VIIA (MyoVIIA) is an unconventional myosin necessary for vertebrate audition [1]–[5]. Human auditory transduction occurs in sensory hair cells with a staircase-like arrangement of apical protrusions called stereocilia. In these hair cells, MyoVIIA maintains stereocilia organization [6]. Severe mutations in the Drosophila MyoVIIA orthologue, crinkled (ck), are semi-lethal [7] and lead to deafness by disrupting antennal auditory organ (Johnston's Organ, JO) organization [8]. ck/MyoVIIA mutations result in apical detachment of auditory transduction units (scolopidia) from the cuticle that transmits antennal vibrations as mechanical stimuli to JO. Principal Findings Using flies expressing GFP-tagged NompA, a protein required for auditory organ organization in Drosophila, we examined the role of ck/MyoVIIA in JO development and maintenance through confocal microscopy and extracellular electrophysiology. Here we show that ck/MyoVIIA is necessary early in the developing antenna for initial apical attachment of the scolopidia to the articulating joint. ck/MyoVIIA is also necessary to maintain scolopidial attachment throughout adulthood. Moreover, in the adult JO, ck/MyoVIIA genetically interacts with the non-muscle myosin II (through its regulatory light chain protein and the myosin binding subunit of myosin II phosphatase). Such genetic interactions have not previously been observed in scolopidia. These factors are therefore candidates for modulating MyoVIIA activity in vertebrates. Conclusions Our findings indicate that MyoVIIA plays evolutionarily conserved roles in auditory organ development and maintenance in invertebrates and vertebrates, enhancing our understanding of auditory organ development and function, as well as providing significant clues for future research. PMID:18461180

  7. The effects of food shortage during larval development on adult body size, body mass, physiology and developmental time in a tropical damselfly.

    PubMed

    Jiménez-Cortés, J Guillermo; Serrano-Meneses, Martín Alejandro; Córdoba-Aguilar, Alex

    2012-03-01

    Few studies have looked jointly at the effects of larval stressors on life history and physiology across metamorphosis, especially in tropical insects. Here we investigated how the variation of food availability during the larval stage of the tropical and territorial American rubyspot damselfly (Hetaerina americana) affects adult body size and body mass, and two physiological indicators of condition--phenoloxidase activity (an indicator of immune ability) and protein concentration. We also investigated whether larval developmental time is prolonged when food is scarce, an expected situation for tropical species whose larval time is less constrained, compared to temperate species. Second instar larvae were collected from their natural environments and reared in one of two diet regimes: (i) "rich" provided with five Artemia salina prey every day, and (ii) "poor" provided with two A. salina prey every day. In order to compare how distinct our treatments were from natural conditions, a second set of last-instar larvae were also collected and allowed to emerge. Only body size and phenoloxidase increased in the rich regime, possibly to prioritize investment on sexually selected traits (which increase mating opportunities), and immune ability, given pathogen pressure. The sexes did not differ in body size in relation to food regimes but they did differ in body mass and protein concentration; this can be explained on the basis of the energetically demanding territorial activities by males (for the case of body mass), and female allocation to egg production (for the case of protein). Finally, animals delayed larval development when food was scarce, which is coherent for tropical environments. These findings provide key insights in the role of food availability in a tropical species. PMID:22085821

  8. Drosophila Wnt and STAT Define Apoptosis-Resistant Epithelial Cells for Tissue Regeneration after Irradiation.

    PubMed

    Verghese, Shilpi; Su, Tin Tin

    2016-09-01

    Drosophila melanogaster larvae irradiated with doses of ionizing radiation (IR) that kill about half of the cells in larval imaginal discs still develop into viable adults. How surviving cells compensate for IR-induced cell death to produce organs of normal size and appearance remains an active area of investigation. We have identified a subpopulation of cells within the continuous epithelium of Drosophila larval wing discs that shows intrinsic resistance to IR- and drug-induced apoptosis. These cells reside in domains of high Wingless (Wg, Drosophila Wnt-1) and STAT92E (sole Drosophila signal transducer and activator of transcription [STAT] homolog) activity and would normally form the hinge in the adult fly. Resistance to IR-induced apoptosis requires STAT and Wg and is mediated by transcriptional repression of the pro-apoptotic gene reaper. Lineage tracing experiments show that, following irradiation, apoptosis-resistant cells lose their identity and translocate to areas of the wing disc that suffered abundant cell death. Our findings provide a new paradigm for regeneration in which it is unnecessary to invoke special damage-resistant cell types such as stem cells. Instead, differences in gene expression within a population of genetically identical epithelial cells can create a subpopulation with greater resistance, which, following damage, survive, alter their fate, and help regenerate the tissue. PMID:27584613

  9. Release of hyaluronidase during in vitro development of Ascaris suum from the third to fourth larval stage.

    PubMed

    Rhoads, M L; Fetterer, R H; Urban, J F

    2001-09-01

    An enzyme that degraded glycosaminoglycan hyaluronic acid was released during in vitro development of Ascaris suum L3 to L4. The enzyme did not hydrolyze glycosaminoglycan chondroitin sulfate A. One molecular form of hyaluronidase was detected, with a molecular weight estimated at 47.8 +/- 8.6 kDa by sucrose density gradient centrifugation and at 55.0 +/- 1.3 kDa by substrate SDS-PAGE zymography. Activity of the enzyme was optimal between pH 5.0 and 6.0, and was present at neutral pH. Hyaluronidase activity was not affected by 5 mM concentrations of cupric sulfate, zinc chloride, calcium chloride, manganese chloride or EDTA. In addition, NaCl had no effect on enzyme activity at concentrations of 0.2-1.0 M. The highest level of hyaluronidase was present in culture fluid collected between days 4 and 6 of in vitro culture, and this period corresponded with that of the highest rate of increase in the percentage of L4. The presence or absence of hyaluronic acid plays a key role in basic developmental processes of vertebrates and is regulated, in part, by hyaluronidases. Developmental processes occurring during the transition of A. suum L3 to L4 may likewise depend on hyaluronidase. In addition, the infection process of a number of organisms, including some nematodes, depends on hyaluronidase. A. suum may likewise utilize hyaluronidase to facilitate larval migration within the host. PMID:11570551

  10. Pigeonpea genotypes influence parasitization preference and survival and development of the Helicoverpa armigera larval parasitoid, Campoletis chlorideae.

    PubMed

    Hugar, Shiddalingappa V; Sharma, Hari C; Basavan Goud, Kondikallu

    2014-01-01

    Studies were undertaken to identify pigeonpea, Cajanus cajan (L.) Millspaugh and the wild relative of pigeonpea, Cajanus scarabaeoides (L.) (accession ICPW 125,) genotypes that are hospitable to the pod borer, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larval parasitoid, Campoletis chlorideae Uchida (Hymenoptera: Ichneumonidae) for the management of this pest in pigeonpea based cropping systems. Percentage parasitization of the H. armigera larvae by the C. chlorideae females was greater under no-choice conditions than under multi-choice conditions because of forced parasitization under no-choice conditions. Lowest parasitization was recorded on the wild relative, ICPW 125, which may be due to long nonglandular hairs and low survival of H. armigera larvae. Parasitization of H. armigera larvae was greater under no-choice, dual-choice and/or multi-choice conditions on ICPL 87, ICPL 87119 and ICPL 87091, which are susceptible to H. armigera, than on the pod borer-resistant genotypes ICPL 332WR, ICPL 84060 and ICPB 2042; while survival and development of the parasitoid was better on H. armigera larvae fed on ICPL 87, ICPL 87119, LRG 41, ICP 7035 and ICPL 87091 than on ICPL 332WR, ICPL 84060, ICPB 2042 and ICPW 125. The genotypes ICPL 87, ICPL 87119, LRG 42 and ICPL 87091 that are hospitable to C. chloridae, are better suited for use in integrated pest management to minimize the losses due to H. armigera in pigeonpea. PMID:25110629

  11. Mitonuclear Epistasis for Development Time and Its Modification by Diet in Drosophila.

    PubMed

    Mossman, Jim A; Biancani, Leann M; Zhu, Chen-Tseh; Rand, David M

    2016-05-01

    Mitochondrial (mtDNA) and nuclear genes have to operate in a coordinated manner to maintain organismal function, and the regulation of this homeostasis presents a substantial source of potential epistatic (G × G) interactions. How these interactions shape the fitness landscape is poorly understood. Here we developed a novel mitonuclear epistasis model, using selected strains of the Drosophila Genetic Reference Panel (DGRP) and mitochondrial genomes from within Drosophila melanogaster and D. simulans to test the hypothesis that mtDNA × nDNA interactions influence fitness. In total we built 72 genotypes (12 nuclear backgrounds × 6 mtDNA haplotypes, with 3 from each species) to dissect the relationship between genotype and phenotype. Each genotype was assayed on four food environments. We found considerable variation in several phenotypes, including development time and egg-to-adult viability, and this variation was partitioned into genetic (G), environmental (E), and higher-order (G × G, G × E, and G × G × E) components. Food type had a significant impact on development time and also modified mitonuclear epistases, evidencing a broad spectrum of G × G × E across these genotypes. Nuclear background effects were substantial, followed by mtDNA effects and their G × G interaction. The species of mtDNA haplotype had negligible effects on phenotypic variation and there was no evidence that mtDNA variation has different effects on male and female fitness traits. Our results demonstrate that mitonuclear epistases are context dependent, suggesting the selective pressure acting on mitonuclear genotypes may vary with food environment in a genotype-specific manner. PMID:26966258

  12. Mitonuclear Epistasis for Development Time and Its Modification by Diet in Drosophila

    PubMed Central

    Biancani, Leann M.; Zhu, Chen-Tseh

    2016-01-01

    Mitochondrial (mtDNA) and nuclear genes have to operate in a coordinated manner to maintain organismal function, and the regulation of this homeostasis presents a substantial source of potential epistatic (G × G) interactions. How these interactions shape the fitness landscape is poorly understood. Here we developed a novel mitonuclear epistasis model, using selected strains of the Drosophila Genetic Reference Panel (DGRP) and mitochondrial genomes from within Drosophila melanogaster and D. simulans to test the hypothesis that mtDNA × nDNA interactions influence fitness. In total we built 72 genotypes (12 nuclear backgrounds × 6 mtDNA haplotypes, with 3 from each species) to dissect the relationship between genotype and phenotype. Each genotype was assayed on four food environments. We found considerable variation in several phenotypes, including development time and egg-to-adult viability, and this variation was partitioned into genetic (G), environmental (E), and higher-order (G × G, G × E, and G × G × E) components. Food type had a significant impact on development time and also modified mitonuclear epistases, evidencing a broad spectrum of G × G × E across these genotypes. Nuclear background effects were substantial, followed by mtDNA effects and their G × G interaction. The species of mtDNA haplotype had negligible effects on phenotypic variation and there was no evidence that mtDNA variation has different effects on male and female fitness traits. Our results demonstrate that mitonuclear epistases are context dependent, suggesting the selective pressure acting on mitonuclear genotypes may vary with food environment in a genotype-specific manner. PMID:26966258

  13. Arm-Gal4 inheritance influences development and lifespan in Drosophila melanogaster.

    PubMed

    Slade, F A; Staveley, B E

    2015-01-01

    The UAS-Gal4 ectopic expression system is a widely used and highly valued tool that allows specific gene expression in Drosophila melanogaster. Yeast transcription factor Gal4 can be directed using D. melanogaster transcriptional control elements, and is often assumed to have little effect on the organism. By evaluation of the consequences of maternal and paternal inheritance of a Gal4 transgene under the transcriptional regulation of armadillo control elements (arm-Gal4), we demonstrated that Gal4 expression could be detrimental to development and longevity. Male progeny expressing arm-Gal4 in the presence of UAS-lacZ transgene had reduced numbers and size of ommatidia, compared to flies expressing UAS-lacZ transgene under the control of other Gal4 transgenes. Aged at 25°C, the median life span of male flies with maternally inherited elav-Gal4 was 70 days, without a responding transgene or with UAS-lacZ. The median life span of maternally inherited arm-Gal4 male flies without a responding transgene was 48 days, and 40 days with the UAS-lacZ transgene. A partial rescue of this phenotype was observed with the expression of UAS-lacZ under paternal arm-Gal4 control, having an average median lifespan of 60 days. This data suggests that arm-Gal4 has detrimental effects on Drosophila development and lifespan that are directly dependent upon parental inheritance, and that the benign responder and reporter gene UAS-lacZ may influence D. melanogaster development. These findings should be taken into consideration during the design and execution of UAS-Gal4 expression experiments. PMID:26505429

  14. Jak-STAT regulation of cyst stem cell development in the Drosophila testis

    PubMed Central

    Sinden, D.; Badgett, M.; Fry, J.; Jones, T.; Palmen, R.; Sheng, X.; Simmons, A.; Matunis, E.; Wawersik, M.

    2012-01-01

    Establishment and maintenance of functional stem cells is critical for organ development and tissue homeostasis. Little is known about the mechanisms underlying stem establishment during organogenesis. Drosophila testes are among the most thoroughly characterized systems for studying stem cell behavior, with germline stem cells (GSCs) and somatic cyst stem cells (CySCs) cohabiting a discrete stem cell niche at the testis apex. GSCs and CySCs are arrayed around hub cells that also comprise the niche and communication between hub cells, GSCs, and CySCs regulates the balance between stem cell maintenance and differentiation. Recent data has shown that functional, asymmetrically dividing GSCs are first established at ~23 hrs after egg laying during Drosophila testis morphogenesis (Sheng et al., 2009). This process correlates with coalescence of the hub, but development of CySCs from somatic gonadal precursors (SGPs) was not examined. Here, we show that functional CySCs are present at the time of GSC establishment, and that Jak-STAT signaling is necessary and sufficient for CySC maintenance shortly thereafter. Furthermore, hyper-activation of Jak in CySCs promotes expansion of the GSC population, while ectopic Jak activation in the germline induces GSC gene expression in GSC daughter cells but does not prevent spermatogenic differentiation. Together, these observations indicate that, similar to adult testes, Jak-STAT signaling from the hub acts on both GSCs and CySC to regulate their development and differentiation, and that additional signaling from CySCs to the GSCs play a dominant role in controlling GSC maintenance during niche formation. PMID:23010510

  15. A detailed staging scheme for late larval development in Strongylocentrotus purpuratus focused on readily-visible juvenile structures within the rudiment

    PubMed Central

    2014-01-01

    Background The purple sea urchin, Strongylocentrotus purpuratus, has long been the focus of developmental and ecological studies, and its recently-sequenced genome has spawned a diversity of functional genomics approaches. S. purpuratus has an indirect developmental mode with a pluteus larva that transforms after 1–3 months in the plankton into a juvenile urchin. Compared to insects and frogs, mechanisms underlying the correspondingly dramatic metamorphosis in sea urchins remain poorly understood. In order to take advantage of modern techniques to further our understanding of juvenile morphogenesis, organ formation, metamorphosis and the evolution of the pentameral sea urchin body plan, it is critical to assess developmental progression and rate during the late larval phase. This requires a staging scheme that describes developmental landmarks that can quickly and consistently be used to identify the stage of individual living larvae, and can be tracked during the final two weeks of larval development, as the juvenile is forming. Results Notable structures that are easily observable in developing urchin larvae are the developing spines, test and tube feet within the juvenile rudiment that constitute much of the oral portion of the adult body plan. Here we present a detailed staging scheme of rudiment development in the purple urchin using soft structures of the rudiment and the primordia of these juvenile skeletal elements. We provide evidence that this scheme is robust and applicable across a range of temperature and feeding regimes. Conclusions Our proposed staging scheme provides both a useful method to study late larval development in the purple urchin, and a framework for developing similar staging schemes across echinoderms. Such efforts will have a high impact on evolutionary developmental studies and larval ecology, and facilitate research on this important deuterostome group. PMID:24886415

  16. Developing a new bait for spotted wing Drosophila in organic cherry production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Studies conducted at the USDA Laboratory in Wapato, WA and at Oregon State University were initiated in 2011 to improve the efficacy of an organically-certified formulation of the insecticide spinosad (Entrust®) for control of the spotted wing drosophila, Drosophila suzukii. Our initial approach was...

  17. Mechanical Control of Organ Size in the Development of the Drosophila Wing Disc

    PubMed Central

    Schluck, Thomas; Nienhaus, Ulrike; Aegerter-Wilmsen, Tinri; Aegerter, Christof M.

    2013-01-01

    Control of cessation of growth in developing organs has recently been proposed to be influenced by mechanical forces acting on the tissue due to its growth. In particular, it was proposed that stretching of the tissue leads to an increase in cell proliferation. Using the model system of the Drosophila wing imaginal disc, we directly stretch the tissue finding a significant increase in cell proliferation, thus confirming this hypothesis. In addition, we characterize the growth over the entire growth period of the wing disc finding a correlation between the apical cell area and cell proliferation rate. PACS numbers: 87.19.lx, 87.18.Nq, 87.80.Ek, 87.17.Ee, 87.85.Xd PMID:24204600

  18. Patterned Anchorage to the Apical Extracellular Matrix Defines Tissue Shape in the Developing Appendages of Drosophila

    PubMed Central

    Ray, Robert P.; Matamoro-Vidal, Alexis; Ribeiro, Paulo S.; Tapon, Nic; Houle, David; Salazar-Ciudad, Isaac; Thompson, Barry J.

    2015-01-01

    Summary How tissues acquire their characteristic shape is a fundamental unresolved question in biology. While genes have been characterized that control local mechanical forces to elongate epithelial tissues, genes controlling global forces in epithelia have yet to be identified. Here, we describe a genetic pathway that shapes appendages in Drosophila by defining the pattern of global tensile forces in the tissue. In the appendages, shape arises from tension generated by cell constriction and localized anchorage of the epithelium to the cuticle via the apical extracellular-matrix protein Dumpy (Dp). Altering Dp expression in the developing wing results in predictable changes in wing shape that can be simulated by a computational model that incorporates only tissue contraction and localized anchorage. Three other wing shape genes, narrow, tapered, and lanceolate, encode components of a pathway that modulates Dp distribution in the wing to refine the global force pattern and thus wing shape. PMID:26190146

  19. Eyg and Ey Pax proteins act by distinct transcriptional mechanisms in Drosophila development

    PubMed Central

    Yao, Jih-Guang; Sun, Y Henry

    2005-01-01

    Drosophila has two pairs of Pax genes, ey/toy and eyg/toe, that play different functions during eye development. ey specifies eye fate, while eyg promotes cell proliferation. We have determined the molecular basis for the functional diversity of Eyg and Ey. Eyg and Ey act by distinct transcriptional mechanisms. They use different DNA-binding domains for target recognition. Most interestingly, Eyg acts exclusively as a repressor, whereas Ey is an activator. Several vertebrate Pax proteins are known to switch between activator and repressor activities, but none as repressors only. Eyg may be the first Pax protein as a dedicated repressor. Vertebrates produce a Pax6 isoform, Pax6-5a, differing from Pax6 in DNA-binding properties and functions and structurally similar to Eyg/Toe. We found that Pax6-5a acts as an activator like Ey, but has DNA-binding specificity like Eyg. PMID:15973436

  20. GAL4 enhancer traps that can be used to drive gene expression in developing Drosophila spermatocytes.

    PubMed

    Bunt, Stephanie M; Monk, Adrian C; Siddall, Nicole A; Johnston, Neisha L; Hime, Gary R

    2012-12-01

    The Drosophila testis has proven to be a valuable model organ for investigation of germline stem cell (GSC) maintenance and differentiation as well as elucidation of the genetic programs that regulate differentiation of daughter spermatogonia. Development of germ cell specific GAL4 driver transgenes has facilitated investigation of gene function in GSCs and spermatogonia but specific GAL4 tools are not available for analysis of postmitotic spermatogonial differentiation into spermatocytes. We have screened publically available pGT1 strains, a GAL4-encoding gene trap collection, to identify lines that can drive gene expression in late spermatogonia and early spermatocytes. While we were unable to identify any germline-specific drivers, we did identify an insertion in the chiffon locus, which drove expression specifically in early spermatocytes within the germline along with the somatic cyst cells of the testis. PMID:22926963

  1. The Kinesin-3, Unc-104 Regulates Dendrite Morphogenesis and Synaptic Development in Drosophila

    PubMed Central

    Kern, Jeannine V.; Zhang, Yao V.; Kramer, Stella; Brenman, Jay E.

    2013-01-01

    Kinesin-based transport is important for synaptogenesis, neuroplasticity, and maintaining synaptic function. In an anatomical screen of neurodevelopmental mutants, we identified the exchange of a conserved residue (R561H) in the forkhead-associated domain of the kinesin-3 family member Unc-104/KIF1A as the genetic cause for defects in synaptic terminal- and dendrite morphogenesis. Previous structure-based analysis suggested that the corresponding residue in KIF1A might be involved in stabilizing the activated state of kinesin-3 dimers. Herein we provide the first in vivo evidence for the functional importance of R561. The R561H allele (unc-104bris) is not embryonic lethal, which allowed us to investigate consequences of disturbed Unc-104 function on postembryonic synapse development and larval behavior. We demonstrate that Unc-104 regulates the reliable apposition of active zones and postsynaptic densities, possibly by controlling site-specific delivery of its cargo. Next, we identified a role for Unc-104 in restraining neuromuscular junction growth and coordinating dendrite branch morphogenesis, suggesting that Unc-104 is also involved in dendritic transport. Mutations in KIF1A/unc-104 have been associated with hereditary spastic paraplegia and hereditary sensory and autonomic neuropathy type 2. However, we did not observe synapse retraction or dystonic posterior paralysis. Overall, our study demonstrates the specificity of defects caused by selective impairments of distinct molecular motors and highlights the critical importance of Unc-104 for the maturation of neuronal structures during embryonic development, larval synaptic terminal outgrowth, and dendrite morphogenesis. PMID:23770702

  2. Rab3-GEF Controls Active Zone Development at the Drosophila Neuromuscular Junction123

    PubMed Central

    Bae, Haneui; Chen, Shirui; Roche, John P.; Ai, Minrong; Wu, Chunlai

    2016-01-01

    Abstract Synaptic signaling involves the release of neurotransmitter from presynaptic active zones (AZs). Proteins that regulate vesicle exocytosis cluster at AZs, composing the cytomatrix at the active zone (CAZ). At the Drosophila neuromuscular junction (NMJ), the small GTPase Rab3 controls the distribution of CAZ proteins across release sites, thereby regulating the efficacy of individual AZs. Here we identify Rab3-GEF as a second protein that acts in conjunction with Rab3 to control AZ protein composition. At rab3-GEF mutant NMJs, Bruchpilot (Brp) and Ca2+ channels are enriched at a subset of AZs, leaving the remaining sites devoid of key CAZ components in a manner that is indistinguishable from rab3 mutant NMJs. As the Drosophila homologue of mammalian DENN/MADD and Caenorhabditis elegans AEX-3, Rab3-GEF is a guanine nucleotide exchange factor (GEF) for Rab3 that stimulates GDP to GTP exchange. Mechanistic studies reveal that although Rab3 and Rab3-GEF act within the same mechanism to control AZ development, Rab3-GEF is involved in multiple roles. We show that Rab3-GEF is required for transport of Rab3. However, the synaptic phenotype in the rab3-GEF mutant cannot be fully explained by defective transport and loss of GEF activity. A transgenically expressed GTP-locked variant of Rab3 accumulates at the NMJ at wild-type levels and fully rescues the rab3 mutant but is unable to rescue the rab3-GEF mutant. Our results suggest that although Rab3-GEF acts upstream of Rab3 to control Rab3 localization and likely GTP-binding, it also acts downstream to regulate CAZ development, potentially as a Rab3 effector at the synapse. PMID:27022630

  3. Temporal changes in the bacterial community of animal feces and their correlation with stable fly oviposition, larval development, and adult fitness

    PubMed Central

    Albuquerque, Thais A.; Zurek, Ludek

    2014-01-01

    Stable flies are blood-feeding insects with a great negative impact on animals world wide. Larvae develop primarily in animal manure and bacteria are essential for larval development; however, the principle of this dependence is not understood. We hypothesized that as the microbial community of animal manure changes over time, it plays an important role in stable fly fitness. Two-choice bioassays were conducted using 2 week old horse manure (control) and aging horse manure (fresh to 5 week old) to evaluate the effect of manure age on stable fly oviposition. Our data