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Translational control of development in C. elegans.

Translational control by the 3'untranslated regions (3'UTRs) of mRNAs contributes to important events throughout the development of C. elegans. In oocytes and early embryos, maternal mRNAs are controlled by 3'UTR elements to restrict translation of their protein products to specific blastomeres. Localized translation is probably critical for specifying blastomere identity. In both germline and somatic cells, mRNAs from sex determining genes are translationally repressed by 3'UTR controls. These controls balance the activities that specify male and female cell fates. During larval development, the temporal sequence of cell lineages requires 3'UTR-mediated regulation of heterochronic genes by a small non-protein coding RNA. We review what is known about these translational control mechanisms in C. elegans. This overview illustrates that translational control by 3'UTR elements is a powerful mechanism for regulating the expression of multiple gene products in diverse cell types during development of a multi-cellular animal.Copyright 1997 Academic Press Limited.

Journal Article↗

Neuropeptide Y: localization in the brain and pituitary of the developing frog (Rana esculenta).

The immunohistochemical localization of neuropeptide Y (NPY)-like peptide has been investigated in the peripheral terminal nerve, brain and pituitary of the frog, Rana esculenta, during development. Soon after hatching, a rather simple NPY-immunoreactive (-ir) neuronal system is present, with elements located mainly in the diencephalon. When hind limbs appear and develop, the NPY-neuronal system undergoes considerable elaboration and NPY-ir perikarya appear in several regions of the telencephalon (dorsal, medial, and lateral pallium; medial septum; medioventral telencephalon; anterior preoptic area), diencephalon (ventromedial, central and posterior thalamic nuclei; suprachiasmatic nucleus; infundibulum), mesencephalon (anteroventral mesencephalic tegmentum), and rhombencephalon (central grey; area of the cerebellar and vestibular nuclei). The frequency of NPY-ir neurons increases during larval development, and then decreases in the anterior preoptic area during the metamorphic climax. Dense plexuses of NPY-ir fibers are formed in several brain areas. NPY-ir fibers are found in the peripheral terminal nerve, and ir-neurons through its course along the ventromedial surface of the olfactory bulbs. NPY-ir fiber projections to the median eminence and pars intermedia derive mainly from the ventral infundibular group of NPY-ir neurons, with a contribution from the suprachiasmatic group of NPY neurons. NPY and carboxyl terminal flanking peptide of proneuropeptide Y coexist in the same neurons throughout the brain. The ontogenetic pattern of NPY-ir neuronal system in the brain of Rana esculenta is remarkably different than that reported for Xenopus laevis.

Animals↗

Studies on Onchocerca cervicalis Railliet and Henry 1910: V. The development of Onchocerca cervicalis larvae in the vectors.

Colonies of Culicoides nubeculosus and C; variipennis were established at Winches Farm Field Station, St. Abans. Both species of midge were found to be susceptible to infection with Onchocerca cervicalis microfilariae. Infection was achieved either by engorging through a chick skin membrane on a suspension of microfilariae in blood, or by feeding upon a horse naturally infected with O. cervicalis microfilariae. A large proportion of the microfilariae ingested by the midges penetrated into the haemocoel and migrated first to the thoracic flight muscles and then to the head. Larval development occurred during migration and 3rd stage (infective) larvae of O. cervicalis reached the head of infected midges (kept at 23 degrees C.) 14-15 days after engorgement.

Animals↗

Morphogenesis of Trichostrongylus rugatus and distribution during development in sheep.

Morphogenesis of Trichostrongylus rugatus was examined in 16 sheep experimentally-infected with 120 000 third-stage larvae and killed 2, 4, 6, 8, 10, 12, 16 and 20 days later (DAI). Third stage larvae moulted between Days 4 and 6, and fourth stage larvae moulted on Day 10 after infection. Four sheep first passed eggs in the faeces between Days 16 and 18 after infection. Rate of growth of larvae was constant between 2 and 10 DAI followed by a period of rapid growth from 10 to 16 DAI. Major features of larval development are described. Nematodes were largely restricted to the first 6 m of gut with 71% of worms occurring in the first 3 m.

Abomasum↗

Development of melanin-concentrating hormone-immunoreactive elements in the brain of gilthead seabream (Sparus auratus).

The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.

Animals↗

The biology, ecology, and management of the cat flea.

The cat flea, Ctenocephalides felis felis, is the most important ectoparasite of domestic cats and dogs worldwide. In addition to its annoyance to pets and humans, C. felis felis is responsible for flea bite allergy dermatitis and the transmission of dog tapeworm. The abiotic and biotic factors that affect the development of immature stages are reviewed with special emphasis given to those aspects directly affecting control. Factors influencing host selection and feeding by adults are summarized. Recent studies concerning mating and oviposition, especially as they impact the likelihood of survival by immatures, are discussed. There has been an increase in the number of reports of insecticide resistance in the past ten years. Greater attention has been placed on disrupting larval development in modern IPM programs. The immature stages of the cat flea are extremely susceptible to environmental factors such as temperature and relative humidity and insect growth regulators (IGRs). In recent years, the control of cat fleas has increasingly relied on the use of IGRs applied to the host or to the indoor environment. Finally, we discuss advances in pesticide chemistry that provide tools for better control of adult fleas on the host.

Animals↗

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

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.

Animals↗

Parasites of domestic and wild animals in South Africa. XLV. Helminths of dairy calves on dry-land Kikuyu grass pastures in the Eastern Cape Province.

Successive pairs of approximately 4-month-old Friesland bull calves, raised under worm-free conditions, were exposed to helminth infection for 14 days on dry-land Kikuyu grass pastures at 28-day to monthly intervals, on a coastal farm in a non-seasonal rainfall region of the Eastern Cape Province. With the exception of one pair of calves exposed for 28 days, this procedure was repeated for 28 consecutive months from December 1982 to March 1985. The day after removal from the pastures one calf of each pair was slaughtered and processed for helminth recovery and the other 21 days later. Both members of the last four pairs of calves were killed 21 days after removal from the pastures. Sixteen nematode species were recovered from the calves, and infection with Ostertagia ostertagi was the most intense and prevalent, followed by Cooperia oncophora. The calves acquired the greatest number of nematodes from the pastures from June to October of the first year and from June to August of the second year of the survey. Few worms were recovered from the tracer calves examined from November or December to March or April in each year of the survey. The seasonal patterns of infection with Cooperia spp., Haemonchus placei, Nematodirus helvetianus, Oesophagostomum spp., O. ostertagi and Trichostrongylus axei were all similar and were negatively correlated to atmospheric temperature and evaporation. Slight to moderate arrest in the development of fourth stage larvae occurred from July to September in Cooperia spp., April to July in H. placei, and August to October in O. ostertagi and Trichostrongylus spp. during the first year of the survey. Too few worms were present in the second year to determine a seasonal pattern of arrest. Species survival during the hot and windy summer months appeared to be achieved via a combination of arrested larval development and an ageing residual population of adult worms in the host, and a small extant population of infective larvae on the pastures.

Animal Husbandry↗

Qualitative and quantitative analyses of juvenile hormone and ecdysteroids from the egg to the pupal molt in Trichoplusia ni.

A method was developed to determine in the same extract juvenile hormone and various types of ecdysteroids in precisely staged eggs and larvae of Trichoplusia ni. Ecdysteroids were tentatively identified on the basis of their retention time in ion suppression reversed-phase HPLC and their cross-reactivity with two relatively non-specific, complimentary antibodies, whereas juvenile hormone was identified using reversed-phase HPLC combined with Galleria bioassay. Freshly laid eggs contained low levels of immunoreactive ecdysteroids. Mid-polar ecdysteroids increased in the phase of segmentation (14-18 h) and 1st larval cuticle formation (36-44 h), when 20-hydroxyecdysone and 20,26-dihydroxyecdysone were found to be predominant. Only traces of ecdysone and 26-hydroxyecdysone were seen. Toward hatching ecdysteroids decreased and represented mainly compounds more polar than 20,26-dihydroxyecdysone. In larval development ecdysteroids were low at the beginning of the feeding phases, increased toward cessation of feeding, and reached highest levels 12-15 h before ecdysis. In feeding stages ecdysone and 20-hydroxyecdysone were predominant, whereas in molting stages they were seen together with 20,26-dihydroxyecdysone and 20-hydroxyecdysonoic acid. The juvenile hormone titer was very low in freshly laid eggs and was high (approximately 25 ng/g) in embryos at the stage of 1st larval cuticle formation and eye pigmentation. In eggs we tentatively identified juvenile hormones I and II, whereas in larval stages juvenile hormone II appeared to be the predominant or exclusive juvenile hormone. Its titer fluctuated rapidly and was high in early 1st-instar larvae and again before the molts into the 3rd, 4th, and 5th instar. Highest titers were reached concomitant with the peak in 20-hydroxyecdysone 12-15 h before ecdysis.

Animals↗

Expression pattern of the homeobox protein NKX2-1 in the developing Xenopus forebrain.

Although morphological data suggest that the amphibian forebrain contains similar subdivisions to those observed in birds and mammals, it is presently unclear whether the same patterning mechanisms are conserved among all three classes of tetrapods. Here we report that NKX2-1, a transcription factor that is essential for the ventral patterning of the forebrain in birds and mammals, is expressed in corresponding (homologous) domains in the developing Xenopus forebrain. NKX2-1 expression is restricted to two domains in the amphibian forebrain: (1) a ventral diencephalic domain, with expression limited to hypothalamic structures; and (2) a telencephalic domain, with expression in the medial ganglionic eminence, preoptic area and part of the septum. Thus, the detailed analysis of the distribution of NKX2-1 provides the first unequivocal evidence for distinct progenitor zones within the amphibian forebrain through embryonic and larval development.

Animals↗

Roles for beta(pat-3) integrins in development and function of Caenorhabditis elegans muscles and gonads.

Heterodimeric integrin receptors for extracellular matrix (ECM) play vital roles in bidirectional signaling during tissue development, organization, remodeling, and repair. The beta integrin subunit cytoplasmic domain is essential for transmission of many of these signals and overexpression of an unpaired beta tail in cultured cells inhibits endogenous integrins. Unlike vertebrates, which have at least nine beta subunit genes, the nematode Caenorhabditis elegans expresses only one beta subunit (betapat-3), and a null mutation in this gene causes embryonic lethality. To determine the functions of integrins during larval development and in adult tissues, we have taken a dominant negative approach by expression of an HA-betatail transgene composed of a hemagglutinin (HA) epitope tag extracellular domain connected to the betapat-3 transmembrane and cytoplasmic domains. Expression of this transgene in muscle and gonad, major sites of integrin expression, caused a variety of phenotypes dependent on the level of transgene expression. Abnormalities in body wall and sex muscles led to uncoordinated movement and egg-laying defects. Significant anomalies in migration and pathfinding were caused by tissue-specific expression of HA-betatail in the distal tip cells (DTC), the cells that direct gonad morphogenesis. A pat-3 gene with Tyr to Phe mutations in the cytoplasmic domain was able to rescue pat-3 null animals but also showed DTC migration defects. These results show that betapat-3 plays important roles in post-embryonic organogenesis and tissue function.

Actins↗

Highwire function at the Drosophila neuromuscular junction: spatial, structural, and temporal requirements.

Highwire is a huge, evolutionarily conserved protein that is required to restrain synaptic growth and promote synaptic transmission at the Drosophila neuromuscular junction. Current models of highwire function suggest that it may act as a ubiquitin ligase to regulate synaptic development. However, it is not known in which cells highwire functions, whether its putative ligase domain is required for function, or whether highwire regulates the synapse during development or alternatively sets cell fate in the embryo. We performed a series of transgenic rescue experiments to test the spatial, structural, and temporal requirements for highwire function. We find that presynaptic activity of highwire is both necessary and sufficient to regulate both synapse morphology and physiology. The Highwire RING domain, which is postulated to function as an E3 ubiquitin ligase, is required for highwire function. In addition, highwire acts throughout larval development to regulate synaptic morphology and function. Finally, we show that the morphological and physiological phenotypes of highwire mutants have different dosage and temporal requirements for highwire, demonstrating that highwire may independently regulate the molecular pathways controlling synaptic growth and function.

Animals↗

Differentiation of somatic mitochondria and the structural changes in mtDNA during development of the dicyemid Dicyema japonicum (Mesozoa).

Dicyemids (Mesozoa) are extremely simple multicellular parasites found in the kidneys of cephalopods. Their mitochondria are known to contain single-gene minicircle DNAs. However, it is not known if the minicircles represent the sole form of mitochondrial genome in these organisms. Here we demonstrate that high-molecular-weight (HMW) mtDNA is present in dicyemids. This form of mtDNA is probably limited to germ cells, and has been analyzed by PCR and Southern hybridization. In situ hybridization revealed that mtDNA is initially amplified during early embryogenesis, and then gradually decreases in copy number as larval development proceeds. Furthermore, we demonstrated using BrdU as a tracer that many of the mitochondria in terminally differentiated somatic cells no longer support DNA synthesis. Taking these observations into account, we propose an "amplification-dilution" model for mesozoan mtDNA. "Stem" mitochondria in the germ cells (1) amplify the HMW form of mtDNA in early embryos, followed by minicircle formation via DNA rearrangement, or (2) selectively replicate minicircles from the HMW DNA, concomitantly with the differentiation of the soma. Minicircle formation may itself lead to the loss of replication origins. Thereafter, the minicircles are simply distributed to daughter mitochondria without replication, resulting in the "somatic" mitochondria, which have lost the replicative form of the HMW mtDNA. The change in mtDNA configuration is discussed in relation to mitochondrial differentiation.

Animals↗

The hedgehog-related gene wrt-5 is essential for hypodermal development in Caenorhabditis elegans.

The Caenorhabditis elegans genome encodes a series of hedgehog-related genes, which are thought to have evolved and diverged from an ancestral Hh gene. They are classified into several families based on their N-terminal domains. Here, we analyze the expression and function of a member of the warthog gene family, wrt-5, that lacks the Hint/Hog domain. wrt-5 is expressed in seam cells, the pharynx, pharyngeal-intestinal valve cells, neurons, neuronal support cells, the excretory cell, and the reproductive system. WRT-5 protein is secreted into the extracellular space during embryogenesis. Furthermore, during larval development, WRT-5 protein is secreted into the pharyngeal lumen and the pharyngeal expression changes in a cyclical manner in phase with the molting cycle. Deletion mutations in wrt-5 cause embryonic lethality, which are temperature sensitive and more severe at 15 degrees C than at 25 degrees C. Animals that hatch exhibit variable abnormal morphology, for example, bagging worms, blistering, molting defects, or Roller phenotypes. We examined hypodermal cell junctions using the AJM-1Colon, two colonsGFP marker in the wrt-5 mutant background and observed cell boundary abnormalities in the arrested embryos. AJM-1Colon, two colonsGFP protein is also misplaced in pharyngeal muscle cells in the absence of WRT-5. In conclusion, we show that wrt-5 is an essential gene that - despite its lack of a Hint domain - has multiple functions in C. elegans and is implicated in cell shape integrity.

Alleles↗

Spook and Spookier code for stage-specific components of the ecdysone biosynthetic pathway in Diptera.

Ecdysteroids regulate many key developmental events in arthropods including molting and metamorphosis. Recently, members of the Drosophila Halloween group of genes, that are required for embryonic viability and cuticle deposition, have been shown to code for several cytochrome P450 enzymes that catalyze the terminal hydroxylation steps in the conversion of cholesterol to the molting hormone 20-hydroxyecdysone. These P450s are conserved in other insects and each is thought to function throughout development as the sole mediator of a particular biosynthetic step since, where analyzed, each is expressed at all stages of development and shows no closely related homolog in their respective genomes. In contrast, we show here that several dipteran genomes encode two novel, highly related, microsomal P450 enzymes, Cyp307A1 and Cyp307A2, that likely participate as stage-specific components of the ecdysone biosynthetic machinery. This hypothesis comes from the observation that Cyp307A1 is encoded by the Halloween gene spook (spo), but unlike other Halloween class genes, Dmspo is not expressed during the larval stages. In contrast, Cyp307a2, dubbed spookier (spok), is expressed primarily during larval stages within the prothoracic gland cells of the ring gland. RNAi mediated reduction in the expression of this heterochromatin localized gene leads to arrest at the first instar stage which can be rescued by feeding the larva 20E, E or ketodiol but not 7dC. In addition, spok expression is eliminated in larvae carrying mutations in molting defective (mld), a gene encoding a nuclear zinc finger protein that is required for production of ecdysone during Drosophila larval development. Intriguingly, mld is not present in the Bombyx mori genome, and we have identified only one spook homolog in both Bombyx and Manduca that is expressed in both embryos and larva. These studies suggest an evolutionary split between Diptera and Lepidoptera in how the ecdysone biosynthetic pathway is regulated during development.

Amino Acid Sequence↗

Oxidation of 3-hydroxykynurenine to produce xanthommatin for eye pigmentation: a major branch pathway of tryptophan catabolism during pupal development in the yellow fever mosquito, Aedes aegypti.

This study concerns the metabolic pathways of 3-hydroxykynurenine in Aedes aegypti mosquitoes during development with emphasis on its oxidation pathway to produce xanthommatin during eye pigmentation. Oxidation of tryptophan to 3-hydroxykynurenine is the major pathway of tryptophan catabolism in Aedes aegypti, but 3-hydroxykynurenine oxidizes easily under physiological conditions, which stimulate the production of reactive oxygen species. Our data show that in Aedes aegypti, the chemically reactive 3-hydroxykynurenine is converted to the chemically stable xanthurenic acid by a transaminase-catalyzed reaction during larval development, while 3-hydroxykynurenine is transported to the compound eyes for eye pigmentation during pupal development. Our data suggest that (1) the transamination pathway of 3-hydroxykynurenine is down-regulated during the pupal development, (2) 3-hydroxykynurenine produced in other body tissues is actively transported to the compound eyes during the pupal stage, (3) the compound eye is the place where ommochromes are produced, and (4) formation of ommochromes results from nonenzymatic oxidation of 3-hydroxykynurenine in the compound eyes.

Aedes↗

Distinct developmental function of two Caenorhabditis elegans homologs of the cohesin subunit Scc1/Rad21.

Cohesin, which mediates sister chromatid cohesion, is composed of four subunits, named Scc1/Rad21, Scc3, Smc1, and Smc3 in yeast. Caenorhabditis elegans has a single homolog for each of Scc3, Smc1, and Smc3, but as many as four for Scc1/Rad21 (COH-1, SCC-1/COH-2, COH-3, and REC-8). Except for REC-8 required for meiosis, function of these C. elegans proteins remains largely unknown. Herein, we examined their possible involvement in mitosis and development. Embryos depleted of the homolog of either Scc3, or Smc1, or Smc3 by RNA interference revealed a defect in mitotic chromosome segregation but not in chromosome condensation and cytokinesis. Depletion of SCC-1/COH-2 caused similar phenotypes. SCC-1/COH-2 was present in cells destined to divide. It localized to chromosomes in a cell cycle-dependent manner. Worms depleted of COH-1 arrested at either the late embryonic or the larval stage, with no indication of mitotic dysfunction. COH-1 associated chromosomes throughout the cell cycle in all somatic cells undergoing late embryogenesis or larval development. Thus, SCC-1/COH-2 and the homologs of Scc3, Smc1, and Smc3 facilitate mitotic chromosome segregation during the development, presumably by forming a cohesin complex, whereas COH-1 seems to play a role important for development but unrelated to mitosis.

Animals↗

Bacterially produced dsRNA targeting SePGRP-LB reduces population fitness of Spodoptera exigua (Lepidoptera: Noctuidae) and increases its susceptibility to SeMNPV.

The beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), is an important agricultural pest, and S. exigua multiple nucleopolyhedrovirus (SeMNPV) is a host-specific biological control agent. However, baculovirus efficacy can be limited by host antiviral responses. S. exigua peptidoglycan recognition protein LB (SePGRP-LB) has been identified as an antiviral immune factor, suggesting that its suppression may increase larval susceptibility to SeMNPV. In this study, bacterially produced double-stranded RNA targeting SePGRP-LB (bac-dsPGRP-LB) was orally delivered to larvae to induce RNA interference. Feeding bac-dsPGRP-LB reduced SePGRP-LB transcript levels by 24.0% to 65.7% over 7 d. SePGRP-LB knockdown prolonged fifth-instar larval development, reduced female pupal weight, shortened male adult longevity and the oviposition period, and decreased fecundity by approximately 51%. Life table analysis further showed significant reductions in the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) following bac-dsPGRP-LB treatment. During SeMNPV infection, co-feeding with bac-dsPGRP-LB significantly suppressed SePGRP-LB expression, increased the SeMNPV genomic load, and reduced larval survival compared with the SeMNPV + bac-dsGFP treatment. These findings identify SePGRP-LB as a promising RNAi target for simultaneously reducing S. exigua fitness and enhancing its susceptibility to SeMNPV under laboratory conditions.

SePGRP-LB↗