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Accumulation of proto-oncogene c-erb-A related transcripts during Xenopus development: association with early acquisition of response to thyroid hormone and estrogen.

The expression of genes encoding receptors for estrogen and thyroid hormones, as well as total c-erb-A related transcripts was determined in unfertilized eggs, all stages of embryonic and larval development and in adult tissues of Xenopus, by quantitative Northern and slot-blot hybridization. DNA and antisense RNA probes complementary to Xenopus c-myc, cytoskeletal actin and albumin mRNAs served as controls or developmental markers. Hybridization to full-length chicken c-erb-A cDNA at moderate stringency revealed a complex biphasic ontogenic pattern for several c-erb-A related mRNAs in all tissues and at all developmental stages, an increase of 4-fold in the accumulation of these transcripts occurring before metamorphosis (stages 30 to 40-42) and followed by a gradual build-up after mid-metamorphosis (stage 56). Using full-length or ligand-binding domain fragments of thyroid hormone (TR) and estrogen (ER) cDNAs under stringent hybridization conditions, transcripts of TR and ER were detected from stages 44 and 54 onwards, respectively. The alpha and beta forms of TR mRNAs exhibited different patterns of accumulation during development, the former transcript being present in substantially higher amounts at all developmental stages. The distinct patterns of accumulation of TR and ER mRNAs could be correlated with the differential pattern of early developmental acquisition of sensitivity of Xenopus larval tissues to thyroid hormone and estrogen.

Animals↗

Development of early brainstem projections to the tail spinal cord of Xenopus.

Horseradish peroxidase (HRP) was used to determine the sequence in which axons from different brain neurons reach the tail spinal cord during embryonic and early larval development of Xenopus laevis. Brainstem cells of several classes project to the tail at these stages: mesencephalic reticulospinal neurons of the nucleus of the medial longitudinal fasciculus, a variety of other reticulospinal neurons, vestibulospinal neurons, and a group of median basal cells which may be raphe neurons. Among the reticulospinal neurons the paired Mauthner cells are the most prominent. They and caudally situated reticular neurons are the first to label with HRP applied to the tail spinal cord (stage 37). Vestibulospinal and other reticular neurons begin to label next (stage 39), followed by mesencephalic and then median basal neurons (stage 41). Except for the Mauthner cells, the number of labeled cells belonging to each neuron class increases gradually as development proceeds.

Animals↗

IMP-L3, A 20-hydroxyecdysone-responsive gene encodes Drosophila lactate dehydrogenase: structural characterization and developmental studies.

IMP-L3, a gene isolated as a potential mediator of imaginal disc morphogenesis in Drosophila melanogaster, encodes lactate dehydrogenase (LDH). The predicted amino acid sequence of IMP-L3 is 58-61% identical to those of human LDHs. In cultured imaginal discs, IMP-L3 transcript levels and LDH enzyme activity increase in response to the steroid hormone, 20-hydroxyecdysone. In embryos, IMP-L3 transcript and LDH activity appear in developing somatic muscles by late stage 13, well before the onset of muscular contraction. High levels of transcript and LDH activity persist throughout embryogenesis and throughout larval development. The gene has been localized by in situ hybridization and deficiency mapping to 65A7-65B2 on the third chromosome. LDH activity is reduced to approximately 50% of wild type in animals heterozygous for a deficiency that removes the 65A-B region. Embryos deficient for the 65A-b region lack LDH activity. We conclude that IMP-L3 is the only gene that encodes LDH in Drosophila.

Amino Acid Sequence↗

The Caenorhabditis elegans gene ham-2 links Hox patterning to migration of the HSN motor neuron.

The Caenorhabditis elegans HSN motor neurons permit genetic analysis of neuronal development at single-cell resolution. The egl-5 Hox gene, which patterns the posterior of the embryo, is required for both early (embryonic) and late (larval) development of the HSN. Here we show that ham-2 encodes a zinc finger protein that acts downstream of egl-5 to direct HSN cell migration, an early differentiation event. We also demonstrate that the EGL-43 zinc finger protein, also required for HSN migration, is expressed in the HSN specifically during its migration. In an egl-5 mutant background, the HSN still expresses EGL-43, but expression is no longer down-regulated at the end of the cell's migration. Finally, we find a new role in early HSN differentiation for UNC-86, a POU homeodomain transcription factor shown previously to act downstream of egl-5 in the regulation of late HSN differentiation. In an unc-86; ham-2 double mutant the HSNs are defective in EGL-43 down-regulation, an egl-5-like phenotype that is absent in either single mutant. Thus, in the HSN, a Hox gene, egl-5, regulates cell fate by activating the transcription of genes encoding the transcription factors HAM-2 and UNC-86 that in turn individually control some differentiation events and combinatorially affect others.

Amino Acid Sequence↗

The roles of the homeobox genes aristaless and Distal-less in patterning the legs and wings of Drosophila.

In the leg and wing imaginal discs of Drosophila, the expression domains of the homeobox genes aristaless (al) and Distal-less (Dll) are defined by the secreted signaling molecules Wingless (Wg) and Decapentaplegic (Dpp). Here, the roles played by al and Dll in patterning the legs and wings have been investigated through loss of function studies. In the developing leg, al is expressed at the presumptive tip and a molecularly defined null allele of al reveals that its only function in patterning the leg appears to be to direct the growth and differentiation of the structures at the tip. In contrast, Dll has previously been shown to be required for the development of all of the leg more distal than the coxa. Dll protein can be detected in a central domain in leg discs throughout most of larval development, and in mature discs this domain corresponds to the distal-most region of the leg, the tarsus and the distal tibia. Clonal analysis reveals that late in development these are the only regions in which Dll function is required. However, earlier in development Dll is required in more proximal regions of the leg suggesting it is expressed at high levels in these cells early in development but not later. This reveals a correlation between a temporal requirement for Dll and position along the proximodistal axis; how this may relate to the generation of the P/D axis is discussed. Dll is required in the distal regions of the leg for the expression of tarsal-specific genes including al and bric-a-brac. Dll mutant cells in the leg sort out from wild-type cells suggesting one function of Dll here is to control adhesive properties of cells. Dll is also required for the normal development of the wing, primarily for the differentiation of the wing margin.

Alleles↗

Sexual dimorphism in life history plasticity in the gypsy moth (Lymantria dispar L.).

Sexual differences in reaction norms of life history traits (larval development time--LDT, pupal weight--PW and adult longevity--L) were investigated in the gypsy moth reared on young or old oak leaves during the first larval instar. Sexual dimorphism was revealed for genetic variation in reaction norms that was expressed only for LDT in males, and PW and L in females. Higher mean plasticity of longevity was found in males compared to females indicating that the sexes are exposed to divergent selective pressures. Greater dependence of males on energy resources (carbohydrates and lipids) may account for the observed differences.

Adaptation, Physiological↗

A modified filter paper culture technique for screening of Strongyloides stercoralis ivermectin sensitivity in clinical specimens.

An in vitro test for the detection of ivermectin sensitivity of Strongyloides stercoralis in human clinical specimens was developed. The test has a simple procedure that combines parasite culture with a drug assay. It is based on the principle of the drug effect on larval development test, combined with a modified coproculture using the filter paper culture technique. This does not require parasite isolation, and drug sensitivity can be shown in relationship to the dose. This test can be applied in field surveys for anthelmintic sensitivity and for appropriate drug rotation strategies when drug resistance occurs.

Animals↗

Research on the control of bluetongue in livestock by vector suppression.

From 1979-83, research has been conducted along the South Platte River drainage system northeast of Denver, CO, to develop and evaluate techniques of wide-area vector suppression for the control of bluetongue (BT) disease in ruminant livestock. Water management and chemical treatment at larval development sites have resulted in a decrease in the activity of Culicoides variipennis (Coquillett) in the 100 km2 treatment area, and there is some correlation between levels of vector activity (as reflected by light trap catches) and the levels of BT in livestock as measured by serologic and virologic evaluations.

Animals↗

Comparative analysis of the development of the mandibular salivary glands and the labial silk glands in the mulberry silkworm, Bombyx mori.

The mulberry silkworm, Bombyx mori has a pair of salivary glands arising from the mandibular segment, in addition to the labial silk glands which are generally considered as modified salivary glands. Here we report the characterization of salivary glands and the comparative gene expression profiling of the silk and salivary glands. The two independent salivary glands made up by 330 cells, grow about 1000 fold during larval development. These individual glands extend up to the T(1) thoracic segment unlike silk glands with fused anterior ends and extending up to the caudal region. The salivary glands also undergo endomitosis resembling the silk glands. The B. mori homologue of the homeotic gene Deformed (BmDfd) was expressed in the mandibular and maxillary segments in stage 17 embryo and got localized to the centre of the mandibular segment at stage 18 to form the salivary gland placodes. The expression was also seen in the distal ends of the leg appendages after blastokinesis (stage 22). Only low variations in BmDfd expression ranging from 1.6 to 2.1 fold were apparent during embryonic development. BmDfd expression was observed in the salivary glands all through the larval instars but not in the silk glands. The transcription factor, Forkhead and the segment polarity gene, Wingless were expressed throughout the salivary glands, the latter confirming the absence of physiological compartmentation within these glands unlike the silk glands. The expression of Amylase and Fibrohexamerin was restricted to the salivary and silk glands, respectively and therefore, served as molecular markers for these tissues.

Amylases↗

Breeding biology of the intertidal sand crab, Emerita (Decapoda: Anomura).

Emerita is a burrowing mole crab or sand crab, adapted to life in wave-washed sandy beaches of temperate and tropical seas. The reproductive biology of this anomuran crab presents several peculiarities, all contributing to its adaptation to this harsh environmental niche. We discuss the following aspects: 1) sex ratio and size at sexual maturity, 2) neoteny and protandric hermaphroditism, 3) mating behaviour and sperm transfer strategy, 4) synchronisation of moulting and reproduction, 5) environmental impact on reproductive cycle and egg production, 6) biochemistry of yolk utilisation and energetics, 7) larval development, dispersal and settlement and 8) the value of Emerita as indicator species. These aspects are discussed in the light of the life history pattern, comprising a sedentary adult and pelagic larval phases. The successful colonisation of the physically challenging habitat of the sandy beach by Emerita is attributable largely to reproductive strategy and the larval developmental and recruitment pattern. Sensitivity to changing environmental conditions, including pollution, make this intertidal crab an indicator species for monitoring anthropogenic impact.

Animals↗

Interactions between an entomopathogenic microsporidium and the endoparasitoid Glyptapanteles liparidis within their host, the gypsy moth larva.

Interactions in the host-parasitoid-pathogen system, Lymantria dispar L. (Lep., Lymantriidae)-Glyptapanteles liparidis (Bouché) (Hym., Braconidae)-Vairimorpha sp. (Protista, Microspora), were investigated. Host selection experiments revealed that G. liparidis females did not discriminate between infected and uninfected host larvae for oviposition. Transmission of the microsporidium from infected to uninfected hosts by stinging female wasps could not be ascertained. Females that developed in infected L. dispar larvae did not transmit the pathogen via oviposition. Vairimorpha infection of the host negatively affected the performance of the braconid, when inoculation took place either before or after parasitization. Microsporidiosis of the host caused delayed development, reduced pupation and adult eclosion, reduction in size and weight, and reduction of adult longevity of G. liparidis. Parasitoids themselves were not systemically infected by Vairimorpha sp., but braconid larvae did ingest microsporidian spores at the end of their endoparasitic development and accumulated the undigested and ungerminated spores in the blind midgut. Negative effects of host infection on parasitoid larvae were detectable from the beginning of parasitoid larval development. Lethal time was reduced when L. dispar larvae were infected and parasitized, often at the expense of the parasitoid when G. liparidis were unable to complete endoparasitic development before the host died. Intensity of infection, measured as number of spores produced per milligram fresh weight of L. dispar larva, was slightly higher in parasitized and infected hosts than in unparasitized and infected hosts.

Animals↗

Chytridiomycosis impacts predator-prey interactions in larval amphibian communities.

Despite ecologists increasingly recognizing pathogens as playing significant roles in community dynamics, few experimental studies have quantified patterns of disease impacts on natural systems. Amphibians are experiencing population declines, and a fungal pathogen ( Batrachochytrium dendrobatidis; Chytridiomycota) is a suspected causal agent in many declines. We studied the effects of a pathogenic fungus on community interactions between the gray treefrog, Hyla chrysoscelis, and eastern newts, Notophthalmus viridescens. Recent studies have characterized chytridiomycosis as an emerging infectious disease, whose suspected rapid range expansion and widespread occurrence pose a significant risk for amphibian populations worldwide. We reared larvae in outdoor polyethylene experimental tanks and tested the effects of initial larval density, predator presence, and fungal exposure on Hyla recruitment and predator-prey interactions between Hyla and Notophthalmus. Newts reduced treefrog survival, and high intraspecific density decreased metamorphic body mass independent of B. dendrobatidis. The presence of fungi reduced treefrog body mass at metamorphosis by 34%, but had no significant main effect on survival or larval period length. B. dendrobatidis differentially affected larval development in the presence of predators; Hyla developed slower when reared with the pathogen, but only when newts were present. This significant predator-by-pathogen interaction suggests that the impact of chytridiomycosis on larval amphibians may be exacerbated in complex communities. Our data suggest that B. dendrobatidis effects on host life history may be complex and indirect. Direct measurements of the community-level effects of pathogens offer an important opportunity to understand a significant threat to global biodiversity-declining amphibian populations.

Analysis of Variance↗

The epidemiology of bovine ostertagiasis in the north temperate regions of North America.

Ostertagia ostertagi is widely distributed and is the most pathogenic of the parasitic nematodes affecting cattle in this region. Clinical ostertagiasis is seen mainly in calves and yearlings but outbreaks tend to be sporadic; the subclinical disease is of greater importance. Studies on the population dynamics of the free-living stages have shown that infective larvae can survive on pasture over the relatively severe winter conditions encountered in this region but that such pasture contamination declines during the succeeding grazing season and is lost by midsummer. In the host there is gradual acquisition of worms from pasture over the summer period, with relatively high burdens accumulating in the autumn. A dramatic shift in the proportion of adult to immature worms occurs during the autumn (October). By the time calves are housed, the proportion of larvae (mainly L4) is greater than 80%. This relative distribution of adults to larvae continues through the winter months until early spring when there appears to be a shift to a higher proportion of adults, presumably due to resumption of development of the L4. These persistent L4 stages are considered to be undergoing hypobiosis. Spring infections of calves appear to be mainly acquired from larvae that have successfully overwintered on pasture, and availability of these larvae is drastically reduced by midsummer. There appears to be negligible development of the eggs that result from the spring infections until late summer and early autumn. At this time because of favorable climatic conditions there is rapid larval development which can result in heavy infections in susceptible calves in the autumn.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Haemonchus contortus: in vitro drug screening assays with the adult life stage.

Motility and feeding assays were assessed as in vitro systems for screening of novel compounds for anthelmintic activity against adult Haemonchus contortus. The study aimed to develop an assay with the parasitic adult stage of this species that could be used in conjunction with, or as an alternative to, the free-living larval stage screens commonly used for drug discovery with many parasitic nematode species. The feeding assay showed limitations due to the apparent continuation of a significant degree of feeding in worms showing greatly reduced motility in the presence of some drugs. Hence, it appeared most likely that the feeding assay would underestimate the toxicity of these drugs. The motility assay was able to detect toxicity of known anthelmintics, including the 'slow-acting' benzimidazoles. A small-scale screening exercise used the motility assay to detect toxicity towards adult parasites in 10 compounds out of a group of 200 chemicals (selected due to known toxic effects in larval development assays). The motility assay appeared suitable for drug screening against adult H. contortus. The use of the adult stage for drug screening in this way ensures that the drug is toxic towards the parasite life stage to be targeted in vivo. A lack of activity in subsequent in vivo trials could, therefore, be most likely attributable to host pharmacokinetic factors rather than an intrinsic lack of activity of the drug towards the adult parasite.

Animals↗

Capitella sp. I homeobrain-like, the first lophotrochozoan member of a novel paired-like homeobox gene family.

The paired-like class of homeobox genes contains numerous distinct families, many of which have been implicated in a variety of developmental functions. We report the isolation and expression of a gene with high similarity to Drosophila melanogaster homeobrain from the polychaete annelid Capitella sp. I. The homeobrain-like (hbnl) gene is a paired-like gene that contains a conserved homeodomain, octapeptide region, alanine stretches, and an OAR domain. Gene orthology analyses of the homeodomain from CapI-hbnl places this gene in a new family of paired-like homeodomain genes that includes D. melanogaster homeobrain (hbn) and representatives from all major bilaterian clades as well as a cnidarian gene. CapI-hbnl expression is largely restricted to subsets of cells in the brain and eyes during larval development in Capitella sp. I. The earliest expression of CapI-hbnl is in small discrete cell clusters in the cerebral ganglia. This expression persists through late larval developmental stages whereas expression is absent in postmetamorphic juveniles. Outside the brain, expression is present on the ventral side of the larva in two small cell clusters, at the brain/pharyngeal border and in the anterior-most segment. CapI-hbnl shares features of brain expression with hbn, although in contrast to hbn, which is expressed along the length of the ventral nerve cord, CapI-hbnl has a restricted anterior expression pattern. CapI-hbnl represents an important neural marker for characterization of the annelid nervous system.

Amino Acid Sequence↗

The association between body strike and dermatophilosis of sheep under controlled conditions.

The association of dermatophilosis with body strike of sheep caused by Lucilia cuprina was studied in a controlled environment. When sheep with areas of wet normal fleece and areas of wet fleece containing scabs induced by infection of the skin with Dermatophilus congolensis were exposed to gravid L. cuprina the files oviposited only on dermatophilosis-affected areas. In wet dermatophilosis-affected fleece, naturally oviposited L. Cuprina eggs hatched and the larvae developed to the second instar stage. Artificially implanted L cuprina eggs hatched in both wet dermatophilosis-affected fleece and wet normal fleece. In wet dermatophilosis-affected fleece the larvae from implanted eggs developed normally but in wet normal fleece they did not develop past the first instar stage. It was concluded that wet but otherwise uncomplicated dermatophilosis lesions are attractive to L. cuprina and provide sufficient protein to allow larval development to take place.

Actinomycetales Infections↗

Sperm-limited fecundity in nematodes: how many sperm are enough?

The Bateman principle, which holds that oocytes are the limiting gamete in reproduction, is violated in a variety of species. Self-fertilizing hermaphrodites of the nematode Caenorhabditis elegans provide an example of a system in which sperm number limits lifetime reproductive output, in this species due to the protandrous nature of sperm production that in turn delays the onset of fertilization. This reproductive delay forms the basis of a trade-off between generation time and total fecundity, in which sperm number plays a pivotal role. I use an age-structured population model to describe the number of sperm that maximize fitness, given larval development time and rates of gamete production. The model predicts the evolution of sperm numbers that are consistent with empirical data for C. elegans provided that precocious larval sperm production is taken into account. Several testable hypotheses follow from the model regarding how natural selection and environmental variation may influence patterns of sperm production among populations or species with a similar mode of reproduction.

Age Factors↗

Morphogenesis and organogenesis in the regenerating planktotrophic larvae of asteroids and echinoids.

In a previous study, we described complete body regeneration (with organogenesis) following surgical bisection in the planktotrophic larvae of the asteroids Luidia foliolata and Pisaster ochraceus. Here we present further detailed observations of these unique regenerative processes not presented in the previous paper. Furthermore, we describe for the first time complete regeneration following surgical bisection of planktotrophic larvae of the regular echinoid Lytechinus variegatus and the irregular echinoid Dendraster excentricus. Larvae of both asteroids and echinoids displayed a capacity for rapid regeneration regardless of their developmental stage. Within 48 h after bisection, aggregations of mesenchyme cells with pseudopodia were observed at the site of surgical bisection. These cellular aggregations were similar in appearance to the mesenchymal blastemas that form in adult echinoderms prior to their arm regeneration, and to those described in other deuterostomes that undergo regeneration. When asteroid larvae were surgically bisected in the early stages of their development, clusters of mesenchyme cells developed into completely new pairs of coelomic pouches located anterior to the newly regenerated digestive tract. This indicates that cell fate in regenerating asteroid larvae remains indeterminate during early development. In the larvae of P. ochraceus, regardless of the developmental stage at the time of bisection, both the anterior and posterior portions regenerated all their missing organs and tissues. However, the larvae of L. foliolata displayed differential regenerative capacity in bisected larval halves at the late bipinnaria stage. The differences observed may be due to differences in larval development (L. foliolata has no brachiolaria stage), and may have evolutionary implications. In the regular echinoid L. variegatus, both larval portions regenerated into morphologically and functionally normal larvae that were indistinguishable from non-bisected control larvae. The regenerative processes were similar to those we observed in planktotrophic asteroid larvae. Regenerating larvae readily metamorphosed into normal juveniles. In the irregular echinoid D. excentricus, posterior portions of larvae completed regeneration and metamorphosis, but anterior portions regenerated only partially during the 2-week study. Our observations confirm that asteroid and echinoid larvae provide excellent models for studies of regeneration in deuterostomes.

Animals↗