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Protein catabolism in mosquitoes: ureotely and uricotely in larval and imaginal Aedes aegypti.

Catabolism of excess dietary protein by Aedes aegypti was investigated during larval development, during and after metamorphosis. Activity profiles were established for xanthine dehydrogenase (XDH, uricotelic pathway) and arginase (ureotelic pathway). Both enzymes are active at all times during the life-cycle. During the aquatic larval and pupal instars, XDH and arginase activities increase with body size. Maximal activities of these two enzyme systems coincide with the time of metamorphic restructuring.Both enzymes are found in the fatbody tissue: XDH activity is found in 80% of the tissue, while arginase activity is distributed equally between abdominal fatbody and the thorax. This might indicate a role for arginase other than catabolic, such as energy metabolism.Arginase activity is high in the aquatic instars and low in sugar-fed females but increases after blood-feeding. XDH activity, also high in larvae and pupae, increases markedly after a blood meal.Larval excretion is characterized by the ureotelic pathway. The pupae as closed systems excrete neither uric acid nor urea; urate accumulates during larval and pupal periods, is conserved throughout metamorphosis, and is finally voided with the meconium by the teneral imago. This presents a form of transient storage-excretion.

Journal Article↗

Corepressor requirement and thyroid hormone receptor function during Xenopus development.

The biologic role of hormonal activation of nuclear receptors is well established. Only recently, however, has the biologic significance of repression begun to be appreciated. Amphibian metamorphosis is marked by dramatic thyroid hormone induced changes, including de novo morphogenesis, tissue remodeling, and organ resorption through programmed cell death. These changes involve cascades of gene regulation initiated by 3,5,3'-triiodothyronine (T(3)). T(3) functions by regulating gene expression through thyroid hormone receptor (TR). TRs are DNA-binding transcription factors that belong to the steroid hormone receptor superfamily. In the absence of a ligand, TRs can repress gene expression by recruiting corepressor complexes, whereas liganded TRs recruit coactivator complexes for gene activation. Corepressor and coactivator complexes induce chromatin remodeling to mediate TR regulation of transcription. The mechanisms of TR action permit a dual function for TRs during development. In premetamorphic tadpoles, when TRs are expressed and T(3) levels are barely detectable, unliganded TRs repress transcription through corepressor recruitment. This TR-mediated repression of target genes is critical for proper larval development, allowing tadpole growth and acquisition of metamorphic competence. In contrast, during metamorphosis, endogenous T(3) causes TRs to activate gene expression, leading to tadpole transformation. Several results also support a role for corepressors during metamorphosis. Corepressor targeted functions, however, are still speculative but may again involve TRs. The requirement of active gene repression at different stages during amphibian development establishes an important biologic role for corepressors.

Animals↗

The tropical African latrine blowfly, Chrysomya putoria (Wiedemann).

The status of the tropical African latrine blowfly Chrysomya putoria (Wiedemann, 1830) is recognized as distinct from the sub-tropical African blowfly C.chloropyga (Wiedemann, 1818) (Diptera: Calliphoridae). The biology of C.putoria in laboratory culture is described briefly with emphasis on the differences in development found in three strains of this species from Tanzania, Liberia and Brazil. All three strains were reproductively compatible and there was no evidence of F1 hybrid sterility or hybrid breakdown in subsequent generations. Larval development was significantly faster in the Brazilian strain than in the strain from Tanzania. Sexual maturity was significantly faster in adults of the Liberian and Brazilian strains compared to that found in the strain from Tanzania. This latter strain was affected by the source of protein available as adult food. These differences between strains did not appear to be due to selection in laboratory culture, but rather evidence of the evolution of physiological divergence within a species occupying different geographical regions.

Animals↗

Loss of Drosophila borealin causes polyploidy, delayed apoptosis and abnormal tissue development.

The chromosomal passenger complex (CPC) is a key regulator of mitosis in many organisms, including yeast and mammals. Its components co-localise at the equator of the mitotic spindle and function interdependently to control multiple mitotic events such as assembly and stability of bipolar spindles, and faithful chromosome segregation into daughter cells. Here, we report the first detailed characterisation of a CPC mutation in Drosophila, using a loss-of-function allele of borealin (borr). Like its mammalian counterpart, Borr colocalises with the CPC components Aurora B kinase and Incenp in mitotic Drosophila cells, and is required for their localisation to the mitotic spindle. borr mutant cells show multiple mitotic defects that are consistent with loss of CPC function. These include a drastic reduction of histone H3 phosphorylation at serine 10 (a target of Aurora B kinase), a pronounced attenuation at prometaphase and multipolar spindles. Our evidence suggests that borr mutant cells undergo multiple consecutive abnormal mitoses, producing large cells with giant nuclei and high ploidy that eventually apoptose. The delayed apoptosis of borr mutant cells in the developing wing disc appears to cause non-autonomous repair responses in the neighbouring wild-type epithelium that involve Wingless signalling, which ultimately perturb the tissue architecture of adult flies. Unexpectedly, during late larval development, cells survive loss of borr and develop giant bristles that may reflect their high degree of ploidy.

Animals↗

Expression of isoforms of the neural cell adhesion molecule (NCAM) and polysialic acid during the development of the Bufo arenarum olfactory system.

The neural cell adhesion molecule (NCAM), a member of the immunoglobulin superfamily that promotes Ca(2+)-independent cell-cell adhesion, is expressed as various isoforms generated by alternative splicing. In this study, the expression of the 180 kDa isoform (180-NCAM), total NCAM (180, 140 and 120 kDa isoforms) and the polysialic acid moiety of NCAM (PSA) were analyzed during the development of the olfactory system of the toad Bufo arenarum using specific antibodies and immunofluorescence light microscopy. NCAM and PSA were not found in the ectodermal thickening corresponding to the olfactory placode at early larval stage (stage 17), but by stage 19, total NCAM, 180-NCAM and PSA were all expressed in the invaginating olfactory placode at the sites of cell-cell contact and in the differentiating olfactory epithelium. Later, NCAM isoforms and PSA were found also in the primary fibers of the olfactory nerve and in the olfactory bulb. However, the expression of 180-NCAM decreased near the end of larval development and was absent in post-metamorphic and adult animals. In contrast, total NCAM (representing 140 and/or 120 kDa isoforms) and PSA continued to be expressed in olfactory tissues of post-metamorphic and adult animals, consistent with the persistent neural plasticity of this tissue. Because 180-NCAM has been associated with non-proliferating neurons, its down-regulation in post-metamorphic and adult olfactory system may be associated with the regenerative capability and continuous cell turnover documented for this region in adult animals.

Animals↗

Differentially expressed regulatory genes in honey bee caste development.

In the honey bee, an eminently fertile queen with up to 200 ovarioles per ovary monopolizes colony level reproduction. In contrast, worker bees have only few ovarioles and are essentially sterile. This phenotype divergence is a result of caste-specifically modulated juvenile hormone and ecdysteroid titers in larval development. In this study we employed a differential-display reverse transcription (DDRT)-PCR protocol to detect ecdysteroid-regulated gene expression during a critical phase of caste development. We identified a Ftz-F1 homolog and a Cut-like transcript. Ftz-F1 could be a putative element of the metamorphic ecdysone response cascade of bees, whereas Cut-like proteins are described as transcription factors involved in maintaining cellular differentiation states. The downregulation of both factors can be interpreted as steps in the metamorphic degradation of ovarioles in worker-bee ovaries.

Amino Acid Sequence↗

Effects of copper and decreased salinity on survival rate and development of Tridacna gigas larvae.

Giant clams (Family: Tridacnidae) are endangered species distributed in the Indo-Pacific region. In this study, survival rate and development of Tridacna gigas larvae were studied for three days in ambient water (32 psu), copper (2.5 microg Cu(2+) L(-1)), reduced salinities (25 and 20 psu) and the combination of copper and 25 psu salinity. No significant differences were found in larval development between treatments. The survival rates decreased considerably with reduced salinities although the combination of copper and reduced salinity gave synergistic effects. As a consequence, this could limit population growth of giant clams in coastal areas and could also explain the absence of larval settlement on reefs close to harbours or river mouths. More research is needed to understand the basic requirements and stress tolerance in giant clam larvae for reef restoration and other management actions to be successful.

Analysis of Variance↗

nhr-25, the Caenorhabditis elegans ortholog of ftz-f1, is required for epidermal and somatic gonad development.

We have analyzed the expression and function of the Caenorhabditis elegans gene nhr-25, a member of the widely conserved FTZ-F1 family of nuclear receptors. The gene encodes two protein isoforms, only one of which has a DNA binding domain. nhr-25 is transcribed during embryonic and larval development. A nhr-25::GFP fusion gene is expressed in the epidermis, the developing somatic gonad, and a subset of other epithelial cells. RNA-mediated interference indicates a requirement for nhr-25 function during development: disruption of nhr-25 function leads to embryonic arrest due to failure of the epidermally mediated process of embryo elongation. Animals that survive to hatching arrest as misshapen larvae that occasionally exhibit defects in shedding molted cuticle. In addition, somatic gonad development is defective in these larvae. These results further establish the importance of FTZ-F1 nuclear receptors in molting and developmental control across evolutionarily distant phyla.

Amino Acid Sequence↗

Field study on the survival, migration and overwintering of infective larvae of horse strongyles on pasture in central Ukraine.

Experimental studies on the survival of infective stage larvae of horse strongyles and their ability to overwinter on pasture were carried out in central Ukraine (Poltavska oblast). Faecal pats (1.5 kg each) of naturally infected horses were placed on pasture, and samples of faeces and surrounding vegetation (10 g each) were collected each month, excluding the winter months, from November 2002 until April 2004. The number of infective third stage larvae was calculated in each sample and compared with that from the control faecal samples cultivated in the laboratory. In the control samples, the ratio of infective third stage larvae to the initial number of eggs was from 54.7% in June up to 84.2% in November. This ratio depended on the presence of nematophagous fungi growing in the faeces. On pasture, the development of larvae to the infective third stage took approximately 4 weeks in the warm season, from April until September. In October, a percentage of the eggs (25% to EPG value) did not hatch. No larval development was observed in faeces in November. A minute quantity of larvae, about 0.03% of their initial number, was observed to survive on pasture for the 12 months. Migration of infective larvae from the faeces to vegetation was not intensive, between 71% and 89% of larvae remained in the faeces 4 weeks after deposition of the faecal pats, the percentage related to soil humidity in each month. The proportion of larvae successfully surviving during winter appeared to be maximal in faecal pats deposited on pasture in September of the previous year (up to 42.0% of the initial number of larvae). Some larvae were observed surviving winter in soil beneath the faecal pats. The results of the study demonstrated that horse pastures in the central part of Ukraine are never free from the infective third stage larvae of strongyles.

Animals↗

Thyrotropin-releasing hormone immunoreactivity in the brain and the pituitary during Bufo arenarum development.

The ontogeny of the thyrotropin releasing hormone (TRH) neuronal system was evaluated by immunocytochemistry in Bufo arenarum. The first appearance of TRH immunoreactive fibers was at early premetamorphosis. These fibers were found in small numbers and weakly stained in the median eminence and pars nervosa. With the advance of larval development, TRH-like material stained intensely and tended to aggregate in the median eminence, pars nervosa and pars intermedia. At climax stages immunoreactive fibers and perikarya (weakly stained) were also identified in the preoptic area. In adult specimens TRH perikarya and neuronal fibers were found in the preoptic and infundibular nuclei of the hypothalamus and in the amygdala, septum and diagonal band of Broca of the telencephalon. In addition, TRH neuronal fibers and endings were found in the preoptic-hypophyseal tract, the external zone of the median eminence, the pars nervosa and pars intermedia. Fibers were absent in the pars distalis. This study represents the first immunocytochemical demonstration of TRH in Bufo species, and serves as a basis for clarification of the neuroendocrine regulation of metamorphosis.

Age Factors↗

Evaluation of the genotoxicity of municipal sewage effluent using the marine worm Platynereis dumerilii (Polychaeta: Nereidae).

Samples of settled (primary) effluent were collected from a municipal sewage treatment works at Newton Abbot, Devon, UK, a site which discharges primary effluent via long sea pipeline into the English Channel (minimum of 200-fold initial dilution). Sewage samples were collected during the period February-April 1995 and were analysed for standard physico-chemical parameters (ammonia, chemical oxygen demand, conductivity, non-purgeable organic carbon and settled solids). Samples were also tested for cytotoxicity, genotoxicity, and for developmental effects in the embryo-larval stages of the marine worm, Platynereis dumerilii. Exposure to sewage concentrations of > or = 10% (v/v) in seawater at 20 +/- 1 degrees C led to a marked reduction in normal embryo-larval development (7 h EC50 values from 10% to 18% v/v, n = 5). There was also evidence of a simultaneous delay in the cell cycle progression (as determined by sister chromatid differential staining) following embryo-larval exposures to sewage concentrations of > or = 10% (v/v). Following the calculation of the Maximum Tolerated Dose (MTD), based on cytotoxic and developmental effects, cells from the same embryo-larvae were analysed for chromosomal aberrations (CAs). Results were consistent for all samples tested, demonstrating the absence of cytogenetic damage following the in vivo exposure of polychaete embryo-larvae to settled sewage.

Animals↗

Tetraploid induction by inhibiting mitosis I with heat shock, cold shock, and nocodazole in the hard clam Mercenaria mercenaria (Linnaeus, 1758).

Tetraploid induction by inhibiting mitosis I with heat shock (32, 35, and 38 degrees C), cold shock (1, 4, and 7 degrees C), and nocodazole (0.02 to 1.6 mg/L) was investigated in the hard clam Mercenaria mercenaria. All treatments were applied to fertilized eggs about 5 min before the first cell division at 22 to 23 degrees C, and lasted for 10, 15, and 20 min. Three replicates were produced for each treatment with different parents. The ploidy of resultant larvae and juveniles was determined with flow cytometry. Heat shock of 35 and 38 degrees C was effective in inhibiting mitosis I, producing 54% to 89% tetraploid larvae. Heat shock of 32 degrees C accelerated embryonic development without inhibiting mitosis or producing tetraploids. In all heat-shock groups, the survival to D-stage larvae was lower than in controls, suggesting that heat-shock treatments and tetraploidy were detrimental to larval development. At the juvenile stage, survivors from heat-shock groups contained no tetraploids. Cold shocks suspended the first cell division during the treatment, but produced no tetraploids in the 4 degrees C and 7 degrees C treatment groups. Cold shock of 1 degrees C produced 31% tetraploid larvae in one replicate, with none surviving to juvenile stage. Nocodazole inhibited mitosis I at concentrations of 0.04 mg/L or higher, but did not produce tetraploids. This study indicates that heat shock is most effective in inducing tetraploids through mitosis I inhibition, although none of the induced tetraploids survived to juvenile stage.

Animals↗

Damaged DNA binding protein 1 in Drosophila defense reactions.

We have focused attention on functions of Drosophila damaged DNA binding protein 1 (D-DDB1) in Drosophila hematopoiesis and previously reported that its whole body dsRNA over-expression using a GAL4-UAS targeted expression system results in melanotic tumors and complete lethality. Since the lesions appear to arise as a normal and heritable response to abnormal development, forming groups of cells that are recognized by the immune system and encapsulated in melanized cuticle, D-DDB1 appears to be an essential development-associated factor in Drosophila. To probe the possibility that it contributes to hemocyte development, we used a collagen promoter-GAL4 strain to over-express dsRNA of D-DDB1 in Drosophila hemocytes. The D-DDB1 gene silencing caused melanotic tumors and mortality at the end of larval development. Similarly, it interfered with melanization and synthesis of antimicrobial peptides. Transgenic flies with D-DDB1 gene silencing were found to accumulate abnormal large blood cells, reminiscent of human leukemia, suggesting that D-DDB1 has functions in hemocyte development.

Animals↗

The phenology of malaria mosquitoes in irrigated rice fields in Mali.

A field study was carried out in the large-scale rice irrigation scheme of the Office du Niger in Mali to investigate the relation between anopheline mosquito larval development and small-scale differences in irrigation practices, such as water level, irrigation application and irrigation frequency. The objective of the study was to find out if water management can be used as a tool for vector control to reduce the malaria transmission risk. Larvae of Anopheles gambiae s.s.,; the main malaria vector in the study area, developed mostly in the first 6 weeks after transplanting the rice. During rice development, a succession of anopheline species was observed. This was associated with a marked decrease in light intensity reaching the water surface as plant height increased. Minor differences in water management resulted in noticeable variations in larval densities and species composition. A. gambiae s.s. larvae were most abundant during the early growing stages and almost absent in a closed rice crop. Due to improper drainage after harvest, A. gambiae s.s. breeding was soon re-established in fields where small pools of water were retained. The results suggest that larval mosquito habitats in the Office du Niger can be significantly reduced by water management, simultaneous planting and harvesting and proper drainage of fallow fields.

Agriculture↗

Alterations in development, behavior, and physiology in Drosophila larva that have reduced ecdysone production.

We investigated behavior, physiology, sensitivity to exogenous application of ecdysone, and nerve terminal structure for differences between the reduced ecdysone genotype, ecd(1)/ecd(1), and wild-type control ecd(1)/TM6B animals during the early and late third instars when raised at 25 degrees C. The ecd(1) mutants were able to survive through larval development and form pupae. However, the results demonstrate that the time to pupation is lengthened by about 50 h for the ecd(1)/ecd(1) as compared with the wild-type control siblings. In addition to the lengthened larval cycle in the mutant, ecd(1)/ecd(1) animals, they also display behavioral differences as compared with controls. The rate of body wall contraction and mouth hook movements are reduced in the early third instar of ecd(1)/ecd(1) as compared with controls. The physiological measure of excitatory junction potential amplitude for the combined Is and Ib terminals did not reveal any differences among the two genotypes during the early third instar but the synaptic strength is reduced in the late third instars for controls. Application of exogenous ecdysone is still effective during the late third instar for the ecd(1)/ecd(1) but not the controls. This suggests that endogenous production of ecdysone have already taken place in the wild-type but not the ecd(1)/ecd(1) larvae, thus the rapid nongenomic responses could still be observed in the late third ecd(1)/ecd(1) larvae. Structurally the number of varicosities and the terminal length showed significant differences between ecd(1)/ecd(1) and the wild-type ecd(1)/TM6B genotype in the late third instars.

Animals↗

Sequence and expression of two regulated transcription units during Drosophila melanogaster development: Deb-A and Deb-B.

Two transcription units, Deb-A and Deb-B (developmental embryonic), are present in a recombinant bacteriophage clone containing a DNA insert from Drosophila melanogaster. The insert maps cytologically to chromosome 2R at 48EF. Sequence analysis of the open reading frames indicates that both are putative membrane proteins with similar hydropathy profiles. The expression of the encoded poly(A)+ RNAs is regulated during oocyte embryonic and larval development. The 600 base Deb-A RNA is present in oocytes and increases in abundance until 15 h of embryogenesis. Its abundance level drops dramatically between 15 and 19 h and is modulated at a low level in the three larval instars and pupae. The 525 base Deb-B RNA is present in oocytes at a 10-fold lower level than the Deb-A transcript. Its abundance increases until a maximum level is reached at 9-12 h. After this time the transcript is undetectable on Northern transfers. Both RNAs are present in greater than 95% polyadenylated form in the cytoplasm. Deb-B RNA is found on polysomes in proportions similar to the bulk of poly(A)+ RNA populations. Deb-A RNA, on the other hand is mostly nonpolysomal during embryogenesis. The RNAs from both Deb-A and Deb-B are found uniformly distributed throughout the embryo during development.

Amino Acid Sequence↗

NANOS-3 and FBF proteins physically interact to control the sperm-oocyte switch in Caenorhabditis elegans.

BACKGROUND: The Caenorhabditis elegans FBF protein and its Drosophila relative, Pumilio, define a large family of eukaryotic RNA-binding proteins. By binding regulatory elements in the 3' untranslated regions (UTRs) of their cognate RNAs, FBF and Pumilio have key post-transcriptional roles in early developmental decisions. In C. elegans, FBF is required for repression of fem-3 mRNA to achieve the hermaphrodite switch from spermatogenesis to oogenesis. RESULTS: We report here that FBF and NANOS-3 (NOS-3), one of three C. elegans Nanos homologs, interact with each other in both yeast two-hybrid and in vitro assays. We have delineated the portions of each protein required for this interaction. Worms lacking nanos function were derived either by RNA-mediated interference (nos-1 and nos-2) or by use of a deletion mutant (nos-3). The roles of the three nos genes overlap during germ-line development. In certain nos-deficient animals, the hermaphrodite sperm-oocyte switch was defective, leading to the production of excess sperm and no oocytes. In other nos-deficient animals, the entire germ line died during larval development. This germ-line death did not require CED-3, a protease required for apoptosis. CONCLUSIONS: The data suggest that NOS-3 participates in the sperm-oocyte switch through its physical interaction with FBF, forming a regulatory complex that controls fem-3 mRNA. NOS-1 and NOS-2 also function in the switch, but do not interact directly with FBF. The three C. elegans nanos genes, like Drosophila nanos, are also critical for germ-line survival. We propose that this may have been the primitive function of nanos genes.

Amino Acid Sequence↗

Effects of MON810 Bt field corn on adult emergence of Helicoverpa zea (Lepidoptera: Noctuidae).

A 3-yr study (1996-1998) was conducted to evaluate the effects of MON810 Bt corn on Helicoverpa zea (Boddie) emergence and to determine whether delayed larval development as a result of Bt intoxication results in higher levels of diapause induction and pupal mortality. In the 1997 study, there was no difference in prepupal mortality between corn types, although significantly more prepupae from Bt plots than from non-Bt plots died in emergence buckets before constructing pupal chambers in 1998. In all years, significantly fewer moths emerged from prepupae collected from Bt plots, suggesting that effects of the expressed Cry1Ab extended to the prepupal and pupal stages. Late plantings of corn showed the greatest reductions in moth emergence from Bt corn because environmental conditions were more conducive to trigger diapause at the time H. zea was developing in these plantings. This was supported by a significantly greater proportion of diapausing pupae remaining in the ground in the late plantings of both Bt and non-Bt corn. For April and early May plantings, larval feeding on Bt corn delayed the time to pupation, although there was no significant difference in moth emergence between corn types for those larvae that successfully pupated. Although Bt expression had less impact on the proportion of moths emerging, the actual number of moths emerging from Bt corn was significantly reduced because fewer larvae reached pupation. Delays in adult emergence, along with significant reductions in adult emergence from MON810 Bt corn, should reduce the rates of colonization in soybean and other late host crops but may also result in asynchrony of mating between individuals emerging from Bt and non-Bt corn. This, in turn, may contribute to the evolution of resistance to Bt corn.

Animals↗