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PubMed1995

The Drosophila 63F early puff contains E63-1, an ecdysone-inducible gene that encodes a novel Ca(2+)-binding protein.

Pulses of ecdysone at the end of Drosophila larval development dramatically reprogram gene expression as they signal the onset of metamorphosis. Ecdysone directly induces several early puffs in the salivary gland polytene chromosomes that, in turn, activate many late puffs. Three early puffs, at 2B5, 74EF, and 75B, have been studied at the molecular level. Each contains a single ecdysone primary-response gene that encodes a family of widely expressed transcription factors. We report here a molecular characterization of the 63F early puff. Unexpectedly, we have found this locus to be significantly different from the previously characterized early puff loci. First, the 63F puff contains a pair of ecdysone-inducible genes that are transcribed in the larval salivary glands: E63-1 and E63-2. Second, E63-1 induction in late third instar larvae appears to be highly tissue-specific, restricted to the salivary gland. Third, E63-1 encodes a novel Ca(2+)-binding protein related to calmodulin. The discovery of an ecdysone-inducible Ca(2+)-binding protein provides a foundation for integrating steroid hormone and calcium second messenger signaling pathways and generates an additional level for potential regulation of the ecdysone response.

Amino Acid Sequence↗

PubMed1984

Apurinic endonuclease activity remains constant during early Drosophila development.

An endonuclease activity that acts on alkali-labile lesions in x-irradiated PM2 DNA and recognizes apurinic lesions in heat/acid treated DNA has been partially purified from Drosophila melanogaster embryos and its specific activity monitored throughout early development. The enzyme activity also showed a low level of activity on UV-irradiated DNA. The saturation kinetics observed with both x-irradiated and apurinic PM2 DNA substrates were similar. The endonuclease activity exhibited a broad pH optimum between pH 6 and 8.5 and was almost completely inhibited by 100 mM NaCl, 0.1 mM EDTA, 2 mM CaCl2 and 10 mM NEM. The reaction was not completely dependent on the presence of Mg++cation, but optimum activity was obtained at a concentration of 0.1 mM; concentrations greater than 1 mM Mg++ were inhibitory. The specific activity of the apurinic endonuclease, partially purified from several stages of embryonic and early larval development, remained the same. Unfertilized eggs exhibited a reduced level of this presumptive repair activity.

Animals↗

PubMed1997

Genes necessary for C. elegans cell and growth cone migrations.

The migrations of cells and growth cones contribute to form and pattern during metazoan development. To study the mechanisms that regulate cell motility, we have screened for C. elegans mutants defective in the posteriorly directed migrations of the canal-associated neurons (CANs). Here we describe 14 genes necessary for CAN cell migration. Our characterization of the mutants has led to three conclusions. First, the mutations define three gene classes: genes necessary for cell fate specification, genes necessary for multiple cell migrations and a single gene necessary for final positioning of migrating cells. Second, cell interactions between the CAN and HSN, a neuron that migrates anteriorly to a position adjacent to the CAN, control the final destination of the HSN cell body. Third, C. elegans larval development requires the CANs. In the absence of CAN function, larvae arrest development, with excess fluid accumulating in their pseudocoeloms. This phenotype may reflect a role of the CANs in osmoregulation.

Animals↗

PubMed2001

Hormonal control of male horn length dimorphism in Onthophagus taurus (Coleoptera: Scarabaeidae): a second critical period of sensitivity to juvenile hormone.

Male dung beetles (Onthophagus taurus) facultatively produce a pair of horns that extend from the base of the head: males larger than a threshold body size develop long horns, whereas males that do not achieve this size develop only rudimentary horns or no horns at all. Using topical applications of methoprene, we identified a sensitive period during the feeding stage of third (final) instar larvae when application of methoprene shifted the threshold body size for horn expression. Male larvae that received methoprene at this time delayed horn production until they attained a larger threshold body size than acetone-treated control larvae. This new sensitive period occurs earlier than a sensitive period previously reported for male horn regulation, and it coincides with a morph-specific pulse of ecdysteroid secretion described for this species. It appears that male horn expression is influenced by endocrine events at two different periods of larval development. We incorporate these results into an expanded model for the endocrine regulation of male horn expression.

Journal Article↗

PubMed1991

Aedes triseriatus (Diptera: Culicidae) and La Crosse virus. IV. Nutritional deprivation of larvae affects the adult barriers to infection and transmission.

Groups of Aedes triseriatus (Say) were reared either as nutritionally deprived (two regimens) or well fed (one regimen) throughout larval development, and the vector competence of resulting small, normal, and large females was assessed for La Crosse virus. When fed a high dose of virus (4.6 log10/0.025 ml in Vero cell culture), 90% of small Ae. triseriatus females transmitted La Crosse virus to suckling mice compared with 70% of normal and 42% of large females. Among small females, 100% had disseminated infections as did 86% of normal females, whereas only 69% of large females had disseminated infections. All females had infected mesenterons (midguts). When fed a low dose of virus (2.2 log10/0.025 ml in Vero cell culture) in a second experiment, 15% of small females transmitted compared with 0% of large females; 50% of small females developed disseminated infections compared with 16% of large females. mesenteronal infection occurred in 70% of small but only 32% of large females. Electron microscopy of mesenteronal tissues from large and small females revealed physical differences in the basement membranes (basal laminae). The mesenterons of small females had 3-6 laminae (mean thickness of the basement membrane = 0.14 microns) compared with 9-16 laminae (mean thickness of the basement membrane = 0.24 microns) in large females. These morphological differences indicated that the mesenteronal escape barrier, which accounted for the difference in the percentage of small and large females with disseminated infections, may be, in part, a physical barrier that was modified by nutritional deprivation in the larval instars.

Aedes↗

PubMedDate not supplied

Genetic aspects of aflatoxin B1 resistance in Drosophila melanogaster.

Fifteen wild-type laboratory strains of Drosophila melanogaster were tested for egg-adult viability when exposed to larval development in media containing 0.5 and 1.0 ppm aflatoxin B (AFB1). Significant variation among the strains was demonstrated, especially at the 0.5 ppm AFB1 concentration. Two resistant and two sensitive strains were made isogenic and mated in a 4 X 4 diallel system. Results indicate that differences in AFB1 sensitivity are due to genes with additive effects on viability and that nonsystematic effects due to the interaction of cytoplasms and genes of both paternal and maternal origin are present. A chromosome/cytoplasm substitution analysis was performed using one of the sensitive and one of the resistant strains. Results indicate that genes on chromosomes X and 2 contribute to egg-adult viability differences observed for larval growth on media containing 0.5 and 1.0 ppm AFB1. Also, a significant interaction between chromosome 2 and cytoplasm, both from the resistant strain, was observed, resulting in a twofold increase in viability at 0.5 ppm AFB1 when compared to controls. Possible relationships of these findings with those from vertebrate systems are discussed.

Aflatoxin B1↗

PubMed2003

Patterns of natural selection on size at metamorphosis in water frogs.

Strategies for optimal metamorphosis are key adaptations in organisms with complex life cycles, and the components of the larval growth environment causing variation in this trait are well studied empirically and theoretically. However, when relating these findings to a broader evolutionary or ecological context, usually the following assumptions are made: (1) size at metamorphosis positively relates to future fitness, and (2) the larval growth environment affects fitness mainly through its effect on timing of and size at metamorphosis. These assumptions remain poorly tested, because data on postmetamorphic fitness components are still rare. We created variation in timing of and size at metamorphosis by manipulating larval competition, nonlethal presence of predators, pond drying, and onset of larval development, and measured the consequences for subsequent terrestrial survival and growth in 1564 individually marked water frogs (Rana lessonae and R. esculenta), raised in enclosures in their natural environment. Individuals metamorphosing at a large size had an increased chance of survival during the following terrestrial stage (mean linear selection gradient: 0.09), grew faster and were larger at maturity than individuals metamorphosing at smaller sizes. Late metamorphosing individuals had a lower survival rate (mean linear selection gradient: -0.03) and grew more slowly than early metamorphosing ones. We found these patterns to be consistent over the three years of the study and the two species, and the results did not depend on the nature of the larval growth manipulation. Furthermore, individuals did not compensate for a small size at metamorphosis by enhancing their postmetamorphic growth. Thus, we found simple relationships between larval growth and postmetamorphic fitness components, and support for this frequently made assumption. Our results suggest postmetamorphic selection for fast larval growth and provide a quantitative estimate for the water frog example.

Animals↗

PubMed1995

Molecular analysis of the Methoprene-tolerant gene region of Drosophila melanogaster.

Adult functions of juvenile hormone (JH) have been described for Drosophila melanogaster and other dipteran insects, but preadult function for this hormone remains largely unknown in this order of insects. We have identified a mutation of Drosophila, Methoprene-tolerant (Met), which appears to alter JH reception during late larval development. The molecular cloning of Met will be a step toward understanding this gene and possibly identifying a preadult role(s) for JH. Molecular cloning was initiated using the technique of transposon-tagging with a transposable P element. P-element insertional alleles of Met were generated, and genomic libraries were constructed from two of these alleles. From these libraries P-element-bearing clones were isolated that in situ hybridized to the cytogenetic region where Met had been previously localized by genetic methods. Two of the alleles were shown to have complete P-elements inserted in similar, but not identical, locations in the predicted cytogenetic region where Met is located. A late-larval cDNA library was screened to identify transcriptional units in this region, and clones were recovered with homology to a DNA fragment abutting the P-element insertion site. These clones may represent Met cDNA molecules.

Alleles↗

PubMed1995

Essential role of E2F recognition sites in regulation of the proliferating cell nuclear antigen gene promoter during Drosophila development.

We have found sequences similar to the transcription factor E2F recognition site within the Drosophila proliferating cell nuclear antigen (PCNA) gene promoter. These sequences are located at positions -43 to -36 (site I). and -56 to -49 (site II) with respect to the cap site Glutathione S-transferase (GST)-E2F and GST-DP fusion proteins cooperate and bind to the potential E2F sites in the PCNA promoter in vitro. A binding factor(s) to these sequences that has similar binding specificity to that of E2F was detected in nuclear extracts of Drosophila Kc cells. Furthermore, transient expression of target site for the activation coincided with the E2F sites. These results indicate that the PCNA gene is a likely target gene of E2F. Examination of lacZ expression from PCNA-lacZ fusion genes carrying mutations in either or both of two E2F sites introduced into flies by germ line transformation revealed that site II plays a major role in the PCNA promoter activity during embryogenesis and larval development, although both sites are required for optimal promoter activity. However, for maternal expression in ovaries, either one of the two sites is essentially sufficient to direct optimal promoter activity. These results demonstrate, for the first time, an essential role for E2F sites in regulation of PCNA promoter activity during development of a multicellular organism.

Animals↗

PubMed1991

Morphogenesis of adrenergic cells in a frog parasympathetic ganglion.

The development of the parasympathetic cardiac ganglion of the frog Xenopus laevis was marked by the differentiation of a population of adrenergic small intensely fluorescent (SIF) cells. The neural crest contributes precursors for both SIF cells and the well-studied cholinergic cardiac ganglion neurons; this situation provided an opportunity to determine whether morphogenesis of the two cell types was correlated. Accordingly, we examined the initial differentiation, developmental regulation, and territorial domain of cardiac SIF cells for comparison with their cholinergic neuron neighbors. Adrenergic SIF cells were present during the time when the first cholinergic precursors were becoming postmitotic. Although SIF cells were present first, cholinergic neurons differentiated almost 16 times faster during the first week of embryonic and larval development and outnumbered SIF cells at all subsequent stages. Nonetheless, the accumulation of both cell types were correlated, since the ratio of cholinergic neurons to SIF cells remained at approximately 10 to 1 up to adult life. Early in development, SIF cells and cholinergic neurons were clustered in the sinus venosus portion of the atrium. The asymmetric distribution of cholinergic neurons within the atrium was lost but that of the SIF cells was maintained throughout life. These results identify relationships between the morphogenesis of two cell types in an autonomic ganglion and place constraints upon the cellular mechanisms that could produce the cells from their neural crest precursors.

Animals↗

PubMed1989

Ultrastructural study of the host-parasite interface of Moniliformis moniliformis (Archiacanthocephala) in laboratory-infected rats.

The fine structure of the host-parasite interface of Moniliformis moniliformis in rats is described, comprising investigations on the ontogenic development of the presomal tegument, on the lesions caused by the worms, and on the host cellular reactions at the points of attachment of the worm. The presomal tegument of the worm contained more fibrous elements than the metasomal tegument. The sclerotization increased with the ages of the worms and toward the tip of the proboscis. The presomal surface coat was more coarsely structured and osmiophilic than that of the metasoma and was neither continuous with the contents of the tegumental crypts nor supported by lipids from necrotic host tissue. The surface coat occasionally detached from the proboscis, probably due to the activity of the surrounding granulocytes of the host. The proboscis hooks lost their tegumental covering during their larval development. Hooks of all worms from rats were invested with a semiliquid lipid coat, apparently derived from tegumental excretions at the base of the hooks. The invaginated area of excretion around the base of the hooks was rich in endoplasmic reticulum. The hooks seemed to renew their lipid coats at certain intervals by dipping into the excreted lipid. In rats all worms, irrespective of their age, attached superficially, penetrating only the intestinal mucosa and the tunica propria. No fibrous connective tissue was found in the lesions caused by the worms, indicating that they frequently changed their site of attachment. At 3-10 days postinfection the host's defense cells observed in the lesions consisted mainly of granulocytes, whereas plasma cells were the predominant leukocytes in lesions of older infections.

Acanthocephala↗

PubMed1991

Differential effects of photoperiod on development and reproduction in "montane" and "prairie" strains of the Rocky Mountain wood tick (Acari: Ixodidae).

Progeny of Dermacentor andersoni Stiles adults collected from one "montane" and two "prairie" sites in southern Alberta, Canada, were exposed to combinations of 9:15, 11:13 and 13:11 (L:D) daily photoperiods during various stages of their life cycle. Time required for developmental and reproductive events was recorded. Significant differences in mean molting times of fed larvae were observed, yet the differences were small, and mean times were not attributed to photoperiodic regime. Fed nymphal "prairie" ticks had significantly longer developmental rates under decreasing light regimes than in constant or increasing regimes, while "montane" ticks showed no differential response to any of these regimes. Engorged "montane" females in all regimes weighed significantly more than those in either "prairie" strain, and "prairie" strain females in increasing and constant short-day regimes were significantly heavier than those in decreasing and constant long-day regimes. Photoperiod is unimportant in the regulation of fed-larval development, and the magnitude of female engorgement may be nominally favored by short-day and increasing regimes. The differential phenotypic expression of photoperiodic regulation of fed nymphal development in this tick may have resulted from selective pressures involving differences in host availability and moisture conditions in the two geographic regions.

Alberta↗

PubMed2003

The Caenorhabditis elegans ems class homeobox gene ceh-2 is required for M3 pharynx motoneuron function.

Several homeobox genes, for example those of the ems class, play important roles in animal head development. We report on the expression pattern and function of ceh-2, the Caenorhabditis elegans ems/Emx ortholog. CEH-2 protein is restricted to the nuclei of one type of small muscle cell, one type of epithelial cell, and three types of neurons in the anterior pharynx in the head. We have generated a deletion allele of ceh-2 that removes the homeobox. Animals homozygous for this deletion are viable and fertile, but grow slightly slower and lay fewer eggs than wild type. We assayed the function of two types of pharynx neurons that express ceh-2, the pairs M3 and NSM. M3 activity is substantially reduced in electropharyngeograms of ceh-2 deletion mutants; this defect can account for the observed retardation in larval development, as M3 activity is known to be necessary for effective feeding. NSM function and metabolism are normal based on the assays used. All cells that express ceh-2 in wild type are present in the ceh-2 mutant and have normal morphologies. Therefore, unlike other ems/Emx genes, ceh-2 seems to be important for a late differentiation step and not for neuron specification or regional patterning. Because the CEH-2 homeodomain is well conserved, we tested whether ceh-2 can rescue ems(-) brain defects in Drosophila, despite the apparent differences in biological roles. We found that the C. elegans ems ortholog is able to substitute for fly ems in brain development, indicating that sequence conservation rather than conservation of biological function is important.

Animals↗

PubMed2001

Differential effects of a labial mutation on the development, structure, and function of stomach acid-secreting cells in Drosophila melanogaster larvae and adults.

The differentiation of copper cells, which secrete stomach acid in Drosophila larvae, has been shown previously to be sensitive to the labialk3 mutation. Here we found that stomach acid secretion in adults was insensitive to labk3. The basis for this stage-specific effect was elucidated by characterizing the development, structure, and function of the adult midgut. First, we demonstrated by copper-dependent fluorescence and morphology that copper cells were present in the adult stomach. Fine-structure analysis of adult copper cells led to the identification of a previously unrecognized plasma membrane domain: apicolateral contacts between copper cells and their neighbors consisted of smooth septate junctions that were enriched in alphabeta-spectrin and ankyrin. Second, we demonstrated that adult copper cells were present in labk3/labvd1 (conditional/null) adults. The labial protein was expressed in adult labk3/labvd1 copper cells, but not in larvae. Thus the labk3 mutation had a stage-specific effect on midgut labial expression, but did not appear to affect protein function. Surprisingly, stomach acidification was dispensable during larval development, since labk3/labvd1 mutant larvae that lacked midgut acidification developed into fertile adults.

Actins↗

PubMed2001

Thyroid hormones in growth and development of fish.

The thyroid hormones (THs), thyroxine (T(4)) and triiodothyronine (T(3)) are products of the thyroid gland in all vertebrates. Their role in early development and metamorphosis is well established in mammals and amphibians, respectively, and recently several studies in fish have highlighted the importance of THs during flatfish metamorphosis. THs are present in high quantities in fish eggs and are presumably of maternal origin. During embryogenesis the concentration of T(4) and T(3) in the eggs decrease until endogenous production starts. Thyroid hormone receptors (TR) have been isolated from several teleosts and in common with tetrapods two receptor isoforms have been identified, TR alpha and TR beta. Both the receptors are expressed in early embryos and larvae of the Japanese flounder (Paralichthys olivaceus), zebrafish (Danio rerio) and seabream (Sparus aurata) although a different temporal pattern is apparent. The role of THs and TRs in fish embryogenesis, larval development and during metamorphosis will be discussed.

Amino Acid Sequence↗

PubMed1987

Circumferential cells of the developing Rana catesbeiana lumbar spinal cord.

Neural elements in the lumbar enlargement of the developing Rana catesbeiana spinal cord were labelled by placing chips of dessicated horseradish peroxidase (HRP) into various lesions of the spinal cord. Of the elements labelled in the lumbar enlargement, a population of cells circumjacent to the gray matter was seen to be distinct from all others on the basis of their morphology, position and their putative embryonic origin. One cell type not previously described was a large circumferential cell (LCC) with primary processes completely circumscribing the gray matter. The ventral process crosses the midline and ascends or descends in the ventral funiculus. The dorsal primary process was observed to extend to the midline and turn ipsilaterally in a rostro-caudal direction in the dorsal funiculus. LCC's were present at early stages of larval development (stage III, Taylor and Kollros 1946) but could not be labelled in juvenile frogs. LCC's were only observed in the lumbar enlargement and could only be labelled through HRP applications at that level. They receive abundant synaptic input from the ipsilateral lateral funiculus. Possible roles for the LCC in the early function and development of the ranid lumbar spinal cord are discussed.

Aging↗

PubMed1999

Isolation and characterization of the gene coding for Artemia franciscana TATA-binding protein: expression in cryptobiotic and developing embryos.

Genomic and cDNA clones coding for the Artemia franciscana homolog of the TATA box-binding protein (TBP) were isolated. The C-terminal region of the predicted protein displays up to 92% sequence identity with the conserved C-terminal regions of TBPs from other species. The gene is divided in seven exons that expand over a region of 33 kb. The position of the four introns located in the conserved C-terminal region has been compared with those of other species. Two of these introns have been generally conserved during evolution, another is an arthropod specific intron, present in Drosophila melanogaster and A. franciscana, and the other is only conserved between vertebrates and A. franciscana. Primer extension experiments detected several transcription initiation sites. Northern blot analyses showed the presence of four mRNAs of estimated sizes of 6.8, 2.6, 1.6 and 1.1 kb. Except for the low expression of the 6.8 and 2. 6 kb RNAs in encysted embryos, steady-state levels showed little variation during the activation of the encysted embryo and the first steps of embryonic and larval development. The amount of TBP protein expressed in encysted embryos and developing larvae has been analyzed by Western blot. Cryptobiotic embryos contain significant amounts of TBP although the level of expression increased almost twice during the first 20 h of development. The presence of TBP protein in cryptobiotic embryos suggests that TBP does not play, by itself, a critical role in the arrest of transcription characteristic of these resistance forms.

Amino Acid Sequence↗

PubMed1991

Development of the olfactory bulb in the clawed frog, Xenopus laevis: a morphological and quantitative analysis.

The relationship between olfactory axons and the cells of the olfactory bulb during normal development was analyzed to determine whether olfactory afferent axons could play a role in the induction of olfactory bulb formation. The morphology of the olfactory bulb in Xenopus larvae from stages 26 to 58 and in adult frogs was analyzed with light and electron microscopy. Axons were first observed beneath the basal lamina of the neural tube at stages 30 and 32; at stage 32, neurons in this area of the neural tube began to differentiate. Synapses of olfactory axons onto young neuronal processes were observed as early as stages 36 and 38. By stage 44, all of the layers of the olfactory bulb were present. The basic structure of the mature form of the olfactory bulb was apparent as early as stage 48/49 and remained constant throughout late larval life and even into adulthood, with only the size increasing. To determine the numerical relationship between olfactory axons from both main and vomeronasal epithelia and mitral/tufted cells in the main and accessory olfactory bulbs, a quantitative study was also performed in which the number of olfactory axons and the number of mitral/tufted cells were estimated for larval stages (stages 50 to 58) and adults. The number of axons increased with stage, with a 16-fold increase between stage 58 and adulthood. The number of mitral/tufted cells increased with stage, with only a 2.3-fold increase between stage 58 and adults. There is a correlation between axon number and mitral/tufted cell number during larval development that is consistent with the hypothesis that olfactory axons influence olfactory bulb development. The convergence ratio of olfactory axons onto mitral/tufted cells was 5:1 in larvae and increased to 34:1 in adults; this increase probably results in increased olfactory sensitivity in adult frogs.

Animals↗