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PubMed1998

Evolutionary changes in sites and timing of actin gene expression in embryos of the direct- and indirect-developing sea urchins, Heliocidaris erythrogramma and H. tuberculata.

We describe an evolutionary comparison of expression of the actin gene families of two congeneric sea urchins. Heliocidaris tuberculata develops indirectly via a planktonic feeding pluteus that forms a juvenile rudiment after a long period of larval development. H. erythrogramma is a direct developer that initiates formation of a juvenile rudiment immediately following gastrulation. The developmental expression of each actin isoform of both species was determined by in situ hybridization. The observed expression patterns are compared with known expression patterns in a related indirect-developing sea urchin, Strongylocentrotus purpuratus. Comparisons reveal unexpected patterns of conserved and divergent expression. Cytoplasmic actin, CyIII, is expressed in the aboral ectoderm cells of the indirect developers, but is an unexpressed pseudogene in H. erythrogramma, which lacks aboral ectoderm. This change is correlated with developmental mode. Two CyII actins are expressed in S. purpuratus, and one in H. erythrogramma, but no CyII is expressed in H. tuberculata despite its great developmental similarity to S. purpuratus. CyI expression differs slightly between Heliocidaris and Strongylocentrotus with more ectodermal expression in Heliocidaris. Evolutionary changes in actin gene expression reflect both evolution of developmental mode as well as a surprising flexibility in gene expression within a developmental mode.

Actins↗
PubMed2000

Water management for controlling the breeding of Anopheles mosquitoes in rice irrigation schemes in Kenya.

An experiment to assess the impact of intermittent irrigation on Anopheles larval populations, rice yields and water use was conducted in the Mwea rice irrigation scheme in Kenya. Four water regimes including intermittent irrigation were tested in a complete randomized block experimental design. Intermittent irrigation was carried out on a weekly schedule, with flooded conditions from Saturday through Tuesday morning. Larval sampling at each plot was conducted every Monday and prior to draining of intermittently irrigated subplots on Tuesday. All the adult anopheline mosquitoes emerging from larvae collected in the experimental plots were identified as being An. arabiensis. By far the highest numbers of An. arabiensis 1st instar larvae were found in the intermittently irrigated subplots, indicating that the water regime provided the most attractive environment for egg laying. However, the ratio between the 4th and 1st instar larvae in the subplots was only 0.08, indicating very low survival rates. In contrast, the 4th/1st instar ratio for subplots with other water management regimes ranged between 0.27 and 0.68, suggesting a correspondingly higher survival than observed with intermittent irrigation. The total number of 4th instars was almost the same in the intermittently irrigated subplots and the irrigation system normally practised by the farmers. The failure to eliminate larval development up to the 4th instar in the former method was attributed to residual pools of water. Larval abundance fluctuated throughout the 12-week sampling period. The highest larval densities were recorded in the 3 weeks after transplanting the rice seedlings. Afterwards, larval numbers dropped dramatically as the height of rice plants increased. Rice yields at harvest did not show statistically significant differences among subplots with different water regimes. The average yield per hectare ranged from 4.8-5.3 metric tonnes. The average daily water percolation/seepage rate was 3.6 mm and did not significantly differ among different water management regimes. Further research is necessary to, among other things, determine whether rice yields could be increased by having flooded and drained intervals that were different from those used in this study. It would likewise be important to assess on a wider scale the feasibility of implementing intermittent irrigation with respect to farmer acceptance and required changes in irrigation system design and management.

Animals↗
PubMed1999

CaMKII regulates the density of central glutamatergic synapses in vivo.

Synaptic connections undergo a dynamic process of stabilization or elimination during development, and this process is thought to be critical in memory and learning and in establishing the specificity of synaptic connections. The type II calcium- and calmodulin-dependent protein kinase (CaMKII) has been proposed to be pivotal in regulating synaptic strength and in maturation of synapses during development. Here we describe how CaMKII regulates the formation of central glutamatergic synapses in Caenorhabditis elegans. During larval development, the density of ventral nerve cord synapses containing the GLR-1 glutamate receptor is held constant despite marked changes in neurite length. The coupling of synapse number to neurite length requires both CaMKII and voltage-gated calcium channels. CaMKII regulates GLR-1 by at least two distinct mechanisms: regulating transport of GLR-1 from cell bodies to neurites; and regulating the addition or maintenance of GLR-1 to postsynaptic elements.

Animals↗
PubMed2002

Molecular and biochemical characterization of two P450 enzymes in the ecdysteroidogenic pathway of Drosophila melanogaster.

Five different enzymatic activities, catalyzed by both microsomal and mitochondrial cytochrome P450 monooxygenases (CYPs), are strongly implicated in the biosynthesis of ecdysone (E) from cholesterol. However, none of these enzymes have been characterized completely. The present data show that the wild-type genes of two members of the Halloween family of embryonic lethals, disembodied (dib) and shadow (sad), code for mitochondrial cytochromes P450 that mediate the last two hydroxylation reactions in the ecdysteroidogenic pathway in Drosophila, namely the C22- and C2-hydroxylases. When sad (CYP315A1) is transfected into Drosophila S2 cells, the cells metabolize 2-deoxyecdysone (2dE) to E and the [3H]ketotriol (2,22-dideoxyecdysone) to 22-deoxyecdysone. In contrast, dib (CYP302A1) is responsible for the conversion of the [3H]ketotriol to [3H]2dE. When cells are transfected with both dib and sad, they metabolize the [3H]ketotriol to [3H]E in high yield. The expression of sad and dib is concentrated within the individual segments of the developing epidermis when there is a surge of ecdysteroid midway through embryogenesis. This result occurs before the ring gland has developed and suggests that the embryonic epidermis is a site of ecdysteroid biosynthesis. This pattern then diminishes, and, during late embryogenesis, expression of both genes is concentrated in the prothoracic gland cells of the developing ring gland. Expression of dib and sad continues to be localized in this endocrine compartment during larval development, being maximal in both the late second and third instar larvae, about the time of the premolt peaks in the ecdysteroid titer.

Animals↗
PubMed1991

Patterns of E74A RNA and protein expression at the onset of metamorphosis in Drosophila.

Metamorphosis in Drosophila is triggered by a pulse of the steroid hormone ecdysone at the end of larval development. Ecdysone initiates a genetic hierarchy that can be visualized as a series of puffs in the larval salivary gland polytene chromosomes. The E74 gene is responsible for the early ecdysone-inducible puff at position 74EF and encodes two related DNA-binding proteins which appear to play a regulatory role in the hierarchy. Here we describe the spatial and temporal patterns of E74A RNA and protein expression at the onset of metamorphosis. We use in situ hybridization, antibody stains, and western and northern blot analyses to follow E74A expression from its initial appearance as nascent transcripts on the polytene chromosomes, to spliced mRNA, to post-translationally modified nuclear E74A protein. E74A is expressed in a wide variety of late-third instar tissues, suggesting that it plays a broad pleiotropic role in response to the hormone. In early prepupae, when the overall levels of E74A mRNA are decreasing, relatively high levels of E74A RNA persist in the gut, peripodial membranes of the imaginal discs, and proliferation centers of the brain. The spatial distribution of nuclear E74A protein correlates with the RNA distribution with the single exception that no E74A protein can be detected in the proliferation centers of the brain. There is also a temporal discrepancy between E74A mRNA and protein accumulation. The peak of E74A protein induced by the late larval ecdysone pulse follows the peak of E74A mRNA by approximately 2 h. This delay is not seen in 10 h prepupae, when the next pulse of ecdysone induces the simultaneous expression of E74A mRNA and protein. We discuss possible mechanisms for post-transcriptional regulation of E74A expression and suggest that the unusually long and complex 5' leader in the E74A mRNA may regulate its translation.

Animals↗
PubMed2000

Screening for Drosophila proteins with distinct expression patterns during development by use of monoclonal antibodies.

Kc 167 is a cell line established from Drosophila embryonic hemocytes and has been shown to express many extracellular matrix (ECM) and other proteins important during development. We have screened monoclonal antibodies (mAbs) raised against heparin affinity purified proteins from conditioned medium of Kc 167 cells to identify novel proteins with important roles for development. One mAb recognized a protein expressed with temporary and tissue specific patterns during Drosophila embryogenesis and larval development. This approach is an alternative to screening of Expression Sequence Tag (EST) clones by in situ hybridization to initiate reverse genetics. In addition, a number of mAbs recognizing ECM proteins were also identified. These mAbs will be useful for biochemical and cell biological analyses of Drosophila ECM proteins.

Animals↗
PubMed2002

Acutei and chronic toxicity of nickel to marine organisms: implications for water quality criteria.

Acute and chronic toxicity tests were conducted to determine the effects of nickel on three U.S. west coast marine species: a fish (the topsmelt, Atherinops affinis), a mollusk (the red abalone, Haliotis rufescens), and a crustacean (the mysid, Mysidopsis intii). The 96-h median lethal concentration (LC50) for topsmelt was 26,560 microg/L, and the chronic value for the most sensitive endpoint in a 40-d exposure was 4,270 microg/L. The median effective concentration (EC50) for 48-h abalone larval development was 145.5 microg/L, and the chronic value for juvenile growth in a 22-d exposure through larval metamorphosis was 26.43 microgAL. The mysid 96-h LC50 was 148.6 microg/L, and the chronic value for the most sensitive endpoint in a 28-d, whole life-cycle exposure was 22.09 microg/L. The abalone and mysid acute values were lower than other values available in the literature. Acute-to-chronic ratios for nickel toxicity to the three species were 6.220, 5.505, and 6.727, respectively, which were similar to the only other available saltwater value of 5.478 (for Americamysis [Mysidopsis] bahia) and significantly lower than the existing values of 35.58 and 29.86 for freshwater organisms. Incorporation of data from the present study into calculations for water quality criteria would lower the criterion maximum concentration and raise the criterion continuous concentration for nickel.

Animals↗
PubMed1987

Laboratory rearing techniques and adult life table parameters for Anopheles sergentii from Egypt.

Techniques are presented for maintaining colonies of Anopheles sergentii, an important malaria vector in Egypt. Larval development time and survival rates were determined for 3 rearing solutions and 4 temperatures. Under optimal conditions larval survival rates averaged 85%. Mean life expectancy at emergence for mated An. sergentii was 23.3 days under insectary conditions of 27 +/- 2 degrees C, 70-80% R.H. The net reproductive rate, mean generation time and instantaneous rate of increase were respectively, 45.8 females per female per generation, 29.7 days and 0.127. In the context of vector potential for malaria transmission, An. sergentii has a daily survivorship rate of 0.95.

Actuarial Analysis↗
PubMed1982

Development of the tympanal organ in larvae of the migratory locust (Locusta migratoria).

The tympanal organ of the migratory locust acquires its definitive form during larval development. All the receptor cells (90-100) are present in the 1st instar, whereas the differentiation of the tympanum and the cuticular structures it bears proceeds in steps from one instar to the next. The elevated process is the earliest such structure to appear (2nd instar); it is followed by the pyriform vesicle (3rd instar) and folded body (4th instar). The styliform body first appears in the imago. Although the typical arrangement of the receptor cells is already discernible in the 1st instar, some of the attachment sites change during development, the final configuration appearing only in the imago.

Animals↗
PubMed2002

High-level ivermectin resistance in a field isolate of Haemonchus contortus associated with a low level of resistance in the larval stage: implications for resistance detection.

The IVPro isolate of Haemonchus contortus was isolated in 1999 after significant numbers of the parasite survived an ivermectin capsule treatment of grazing sheep acquiring a natural infection at Prospect, NSW, Australia. The isolate shows high-level resistance to ivermectin (faecal egg count is unaffected by ivermectin oral treatment at 0.2mg kg(-1)). The larval LC(50), as assessed by larval development assays (LDAs), is only approximately two-fold higher than several susceptible isolates, making it difficult to detect the resistance using larval LC(50) as an indicator. However, the isolate shows the presence of a small proportion of the population (<20%) able to develop at significantly higher drug concentrations than the susceptible isolates. Hence, if the IVPro and susceptible isolates are compared at the LC(99) level, the IVPro isolate is readily identifiable as resistant. This degree of distinction at the LC(99) allows the IVPro isolate to be identified as resistant by simply observing the highest drug concentration in the development assay at which some larvae develop relative to the susceptible isolates. Assessing the development assay using this criterion allows a distinction between IVPro and the susceptible isolates equivalent to 10-fold differences in drug concentration, greatly increasing the likelihood of detecting the resistant isolate in routine resistance tests. This study highlights the need to examine this aspect of LDAs in order to detect the type of resistance displayed by IVPro H. contortus.

Animals↗
PubMed2001

Are queen Bombus terrestris giant workers or are workers dwarf queens? Solving the 'chicken and egg' problem in a bumblebee species.

In the social bee, Bombus terrestris, the two castes differ in size and physiology, but not in any other morphological and anatomical aspects. The size differences between the castes are the result of longer instar duration in prospective queen larvae. It appears that queen larvae are programmed to have a higher molting weight at the end of the 2nd, 3rd and 4th instars. Calculation of the growth ratio, the ratio between the logarithm of molting weight at two successive instars, revealed that queen larvae have a linear growth ratio over the entire larval development as predicted by Dyar's rule. In the worker larvae, in contrast, linearity of the growth ratio breaks after the second instar, resulting in larval molting at lower weights than expected by Dyar's rule. We therefore suggest that workers' development is abnormally shortened, either by parental manipulation or by adopting a different growth plan in response to the queen's signal.

Animals↗
PubMed2000

Metamorphosis in the summer flounder, Paralichthys dentatus: thyroidal status influences gill mitochondria-rich cells.

Metamorphosis in the summer flounder (Paralichthys dentatus) is mediated by thyroid hormones (TH) and is accompanied by changes in gill mitochondria-rich cells (MRCs) and in salinity tolerance. Altered thyroid status during larval development and metamorphosis in this species influences salinity tolerance, though the influence of any hormone on MRCs of larval marine teleosts is not known. This study characterized the effect of altered thyroid status on MRC intracellular membranes, mitochondria size and ultrastructure, immunoreactive (ir)-Na(+),K(+)-ATPase, and cell size and density during metamorphosis in summer flounder. Inhibition of metamorphosis with thiourea (30 ppm) (TU, an inhibitor of TH synthesis) inhibited changes in MRCs, producing large "larval" type MRCs with weak reactivity to osmium; large, electron-lucent mitochondria; and weak ir-Na(+),K(+)-ATPase. Replacement of TH with TU + thyroxine-Na salt (100 ppb) rescued the fish from developmental inhibition, producing smaller "juvenile" type MRCs with strong reactivity to osmium; smaller, electron-opaque mitochondria; and strong ir-Na(+), K(+)-ATPase. The findings suggest that TH are necessary for MRCs to change from larval to juvenile form during metamorphosis.

Animals↗
PubMed2005

Energy flow and subsidies associated with the complex life cycle of ambystomatid salamanders in ponds and adjacent forest in southern Illinois.

Breeding adults and metamorphosing larval amphibians transfer energy between freshwater and terrestrial ecosystems during seasonal migrations and emergences, although rarely has this been quantified. We intensively sampled ambystomatid salamander assemblages (Ambystoma opacum,A. maculatum, and A. tigrinum) in five forested ponds in southern Illinois to quantify energy flow associated with egg deposition, larval production, and emergence of metamorphosed larvae. Oviposition by female salamanders added 7.0-761.4 g ash-free dry mass (AFDM) year(-1) to ponds (up to 5.5 g AFDM m(-2) year(-1)). Larval production ranged from 0.4 to 7.4 g AFDM m(-2) year(-1) among populations in three ponds that did not dry during larval development, with as much as 7.9 g AFDM m(-2) year(-1) produced by an entire assemblage. Mean larval biomass during cohort production intervals in these three ponds ranged from 0.1 to 2.3 g AFDM m(-2) and annual P/B (production/biomass) ranged from 4 to 21 for individual taxa. Emergent biomass averaged 10% (range = 2-35%) of larval production; larval mortality within ponds accounted for the difference. Hydroperiod and intraguild predation limited larval production in some ponds, but emerging metamorphs exported an average of 70.0+/-33.9 g AFDM year(-1) (range = 21.0-135.2 g AFDM year(-1)) from ponds to surrounding forest. For the three ponds where larvae survived to metamorphosis, salamander assemblages provided an average net flux of 349.5+/-140.8 g AFDM year(-1) into pond habitats. Among all ponds, net flux into ponds was highest for the largest pond and decreased for smaller ponds with higher perimeter to surface area ratios (r2 = 0.94, P<0.05, n = 5). These results are important in understanding the multiple functional roles of salamanders and the impact of amphibian population declines on ecosystems.

Animals↗
PubMed2005

Host regulation and release of parasitism-specific proteins in the system Toxoneuron nigriceps-Heliothis virescens.

The braconid wasp Toxoneuron nigriceps induced qualitative and quantitative changes in the protein composition of the moth Heliothis virescens host hemolymph. Total protein concentration was found to be higher in parasitized host 4 days after parasitism as compared to control hosts, mainly due to changes in a particular group of proteins. Host proteins with a molecular mass of 173 and 72 kDa were found in higher levels in the hemolymph of parasitized larvae as control hosts approached pupation, while an 80 kDa peptide was found in reduced concentration in the hemolymph of parasitized hosts. Levels of these three peptides were maintained throughout parasitoid development, while two of them (173 and 72 kDa) were cleared from the host hemolymph close to pupation. Besides the regulation of host proteins, three parasitism-specific proteins (PSPs) were released into the host hemolymph. Two of them (PSP1-MW=116 kDa, pI=6.3; PSP2-MW=114 kDa, pI=6.2) first appeared in the hemolymph of parasitized hosts soon after pupation of control host and increased in concentration as the parasitoid developed. The third PSP (PSP3-MW=56 kDa, pI=5.8) was produced towards the end of parasitoid larval development, close to parasitoid egression. Database searches based on the amino acid composition and amino terminal sequence of PSP1 and PSP2 did not produce any significant matches, while PSP3 was identified as a putative chitinase. Incubation of host derived tissues, parasitoid larvae and teratocytes in 35S conditioned media suggested PSPs were a product of teratocytes. The role of the regulation of host proteins and release of PSPs by teratocytes for the successful development of T. nigriceps are discussed.

Amino Acid Sequence↗
PubMed1990

Metallothionein Mto gene of Drosophila melanogaster: structure and regulation.

We report the sequence of the Mto gene, one of the two known metallothionein genes of Drosophila melanogaster, and compare its structure with that of the other metallothionein gene, Mtn. The main structural features are the presence of a small intron (61 base-pairs), the presence of four potential MREs (metal regulatory elements) and the absence of a TATA box in the promoter region. Of all metals tested, Hg2+, Cd2+ and Cu2+ are the most efficient ions for inducing an increase in Mto gene transcription. The Mto and Mtn genes are differentially regulated during normal development. Transcription of Mto is detected early in embryogenesis (0 to 3 h) and persists to the third larval instar, while Mtn expression starts later in embryogenesis (12 to 15 h) and is thereafter maintained throughout larval development and adult stages. Sequencing of the Mto protein is in good agreement with the nucleic acid data. Surprisingly, attempts to isolate and characterize the Mtn protein were unsuccessful. Several lines of evidence suggest that this metallothionein is rapidly incorporated after its synthesis into lysosomes, where it would be processed in a way that would not permit its purification. The function of the Mtn protein thus appears to be mainly related to detoxification processes. The pattern of expression of Mto suggests that this gene may be involved in the control of metal homeostasis during development.

Amino Acid Sequence↗
PubMed1986

Permeability changes in the egg-shell of hookworms during development and eclosion.

The development and hatching of Ancylostoma ceylanicum and Ancylostoma tubaeforme eggs in a range of potentially inimical solutions has been examined using time-lapse video micro-recording techniques. In all compounds tested the larvae developed to the active pretzel stage. The inception of activity resulted in an increase in shell permeability to water and small molecules. In anthelmintics of molecular weight (Mr) 400 development continued normally until about the time that control eggs hatched, when there was a marked decrease in larval activity and eclosion did not occur. The addition of a low Mr surface-active agent potentiated the effect of the anthelmintic and caused larval activity to be lost earlier. The calcium inhibitor lanthanum chloride affected larval development rather than eclosion. Large Mr compounds did not affect either development or hatching and the results suggest that the egg-shell may act as a form of sieve, allowing the essential oxygen to reach the developing larva while excluding large potentially dangerous molecules. No appreciable differences were found between the two species of hookworm examined.

Acridine Orange↗
PubMed2002

Hermaphroditic, demasculinized frogs after exposure to the herbicide atrazine at low ecologically relevant doses.

Atrazine is the most commonly used herbicide in the U.S. and probably the world. It can be present at several parts per million in agricultural runoff and can reach 40 parts per billion (ppb) in precipitation. We examined the effects of atrazine on sexual development in African clawed frogs (Xenopus laevis). Larvae were exposed to atrazine (0.01-200 ppb) by immersion throughout larval development, and we examined gonadal histology and laryngeal size at metamorphosis. Atrazine (> or =0.1 ppb) induced hermaphroditism and demasculinized the larynges of exposed males (> or =1.0 ppb). In addition, we examined plasma testosterone levels in sexually mature males. Male X. laevis suffered a 10-fold decrease in testosterone levels when exposed to 25 ppb atrazine. We hypothesize that atrazine induces aromatase and promotes the conversion of testosterone to estrogen. This disruption in steroidogenesis likely explains the demasculinization of the male larynx and the production of hermaphrodites. The effective levels reported in the current study are realistic exposures that suggest that other amphibian species exposed to atrazine in the wild could be at risk of impaired sexual development. This widespread compound and other environmental endocrine disruptors may be a factor in global amphibian declines.

Animals↗
PubMedDate not supplied

Diverse developmental mechanisms contribute to different levels of diversity in horned beetles.

An ongoing challenge to evolutionary developmental biology is to understand how developmental evolution on the level of populations and closely related species relates to macroevolutionary transformations and the origin of morphological novelties. Here we explore the developmental basis of beetle horns, a morphological novelty that exhibits remarkable diversity on a variety of levels. In this study, we examined two congeneric Onthophagus species in which males develop into alternative horned and hornless morphs and different sexes express marked sexual dimorphism. In addition, both species differ in the body region (head vs. thorax) that develops the horn. Using a comparative morphological approach we show that prepupal growth of horn primordia during late larval development, as well as reabsorption of horn primordia during the pupal stage, contribute to horn expression in adults. We also show that variable combinations of both mechanisms are employed during development to modify horn expression of different horns in the same individual, the same horn in different sexes, and different horns in different species. We then examine expression patterns of two transcription factors, Distal-less (Dll) and aristaless (al), in the context of prepupal horn growth in alternative male morphs and sexual dimorphisms in the same two species. Expression patterns are qualitatively consistent with the hypothesis that both transcription factors function in the context of horn development similar to their known roles in patterning a wide variety of arthropod appendages. Our results suggest that the origin of morphological novelties, such as beetle horns, rests, at least in part, on the redeployment of already existing developmental mechanisms, such as appendage patterning processes. Our results also suggest, however, that little to no phylogenetic distance is needed for the evolution of very different modifier mechanisms that allow for substantial modulation of trait expression at different time points during development in different species, sexes, or tissue regions of the same individual. We discuss the implications of our results for our understanding of the evolution of horned beetle diversity and the origin and diversification of morphological novelties.

Animals↗