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Temporal expression and T3 induction of thyroid hormone receptors alpha1 and beta1 during early embryonic and larval development in zebrafish, Danio rerio.

The effects of thyroid hormones on metabolism and development are mediated by thyroid hormone receptors (TRs). We report the cloning and characterization of a TR beta1 cDNA from zebrafish. Southern blot analysis revealed that there is a single genomic locus for the TR beta gene, while the TR alpha gene potentially has two loci. Multiple TR alpha and TR beta transcripts were detected in adult tissues. Using a semiquantitative RT-PCR assay, zygotic expression of TR alpha1 and TR beta1 were shown to occur before the midblastula transition stage. In transiently transfected HeLa cells, TR alpha1 displayed constitutive transactivation in the absence of ligands, which was slightly enhanced by triiodothyronine (T3). The transactivating activity of TR beta1 was strictly ligand-dependent and repressed in the absence of T3. Finally, the T3 induction of TR alpha1 and TR beta1 mRNAs was demonstrated in zebrafish embryos and larvae. The auto-induction of TR alpha1 and TR beta1 may serve a regulatory role during the embryonic and larval development of zebrafish.

Amino Acid Sequence↗

Effects of nitrate and atrazine on larval development and sexual differentiation in the northern leopard frog Rana pipiens.

Pollution from agrochemicals may be contributing to the global decline in amphibian populations. Environmentally relevant concentrations of nitrate and/or atrazine on anuran development and gonadal differentiation were tested. Four replicates of 20 tadpoles per tank (80/treatment) were exposed from Taylor-Kollros stage 2 to 3 to stage 23 to 34 to either 10 mg/L nitrate, 10 microg/L atrazine, a combined exposure of 10 mg/L nitrate plus 10 microg/L atrazine, or untreated controls. No treatment-dependent effects on weight, snout-vent or hind limb length, or time to forelimb emergence were observed. The proportions of females increased in all treatments compared to the controls, especially in the combined treatment (chi2 = 17.90, df = 6, p = 0.0065, combined = 66.4% female, control = 41% female). The frequency of intersex was low in all treatments. No treatment-related effects on the total number of spermatogenic cells were observed, but the ratio of cell types differed in that testes from animals in the treated groups exhibited more spermatogonia, fewer spermatocytes, and more spermatids than the control (significantly different, Kruskal-Wallis, p < 0.05). Ovaries from animals treated with nitrate or atrazine exhibited larger immature (previtellogenic) and mature (vitellogenic) follicles, but ovaries from the combined treatment had larger immature follicles only. Testicular oocytes were observed in the nitrate-only and atrazine-only treatments, and the control treatment, but not the combined treatment. Overall, this study has demonstrated changes in sex ratios that are more marked in response to combined nitrate/atrazine exposure than with these chemicals alone. Histological evidence suggests that premature maturation of gonad may occur as a result of nitrate and/or atrazine exposure during larval development.

Animals↗

Pine processionary caterpillar allergenicity increases during larval development.

BACKGROUND: In European countries, pine processionary caterpillar (Thaumetopoea pityocampa) is one of the species of Lepidoptera able to affect humans. Its allergenicity has been demonstrated in previous studies. However, in all of them larval extract in the fifth stage of development has been used as antigen. Therefore, allergenicity of other larval stages was unknown. The objective of our study was to detect allergenic changes of pine processionary caterpillar during larval development. METHODS: Pine processionary caterpillar extracts in all larval stages were analyzed by SDS-PAGE. Immunoblotting was performed using patient sera with suggestive history of allergy and a positive prick test.

Allergens↗

Loss of yolk platelets and yolk glycoproteins during larval development of the sea urchin embryo.

The fate of the yolk platelets and their constituent yolk glycoproteins was studied in Strongylocentrotus purpuratus eggs and embryos cultured through the larval stage. Previous studies have shown that the yolk glycoproteins undergo limited proteolysis during early embryonic development. We present evidence that the yolk glycoproteins stored in the yolk platelets exist as large, disulfide-linked complexes that are maintained even after limited proteolysis have occurred. We provide additional evidence that acidification of the yolk platelet may activate a latent thiol protease in the yolk platelet that is capable of correctly processing the major yolk glycoprotein into the smaller yolk glycoproteins. Because we previously showed that these yolk glycoproteins are not catabolized during early embryonic development, it was of interest to study their fate during larval development. Using a specific polyclonal antibody to a yolk glycoprotein, we found that both yolk glycoproteins and the yolk platelets disappeared in feeding, Day 7, larval stage embryos, but that starvation did not significantly affect the levels of the yolk glycoproteins. We also found that the yolk glycoproteins reappeared in 30-day-old premetamorphosis larvae.

Animals↗

Effects of the dinoflagellate Karenia brevis on larval development in three species of bivalve mollusc from Florida.

The effects of Karenia brevis (Wilson clone) on larval survival and development of the northern quahog, Mercenaria mercenaria, eastern oyster, Crassostrea virginica and bay scallop, Argopecten irradians, were studied in the laboratory. Larvae were exposed to cultures of whole and lysed cells, with mean total brevetoxin concentrations of 53.8 and 68.9 microgL(-1), respectively. Survival of early (3-day-old) larvae was generally over 85% for all shellfish species at K. brevis densities of 100 cells ml(-1) or less, and not significantly different between whole and lysed culture. At 1000 cells ml(-1), survival was significantly less in lysed culture than whole culture for both M. mercenaria and C. virginica. Survival of late (7-day-old) larvae in all three species was not significantly affected by K. brevis densities of 1000 cells ml(-1) or less. At 5000 cells ml(-1), however, survival was reduced to 37%, 26% and 19% for A. irradians, M. mercenaria and C. virginica, respectively. Development of C. virginica and M. mercenaria larvae was protracted at K. brevis densities of 1000 cells ml(-1). These results suggest that blooms of K. brevis, and particularly their associated brevetoxins, may have detrimental consequences for Florida's shellfisheries by disrupting critical larval processes. Special attention should be paid to blooms of K. brevis where these shellfish occur naturally or where aquaculture and restoration activities are either ongoing or planned.

Animals↗

Avermectin inhibition of larval development in Haemonchus contortus--effects of ivermectin resistance.

Avermectin (AVM) inhibition of the development of the free-living stages of Haemonchus contortus has been quantified in an assay in which nematode eggs are placed on an agar matrix containing serial dilutions of a drug in the wells of a microtitre plate. Development is allowed to proceed for 6 days by which time larvae in control wells (no drug) have reached the infective, third (L3) stage. At high concentrations (> 30 nM) ivermectin (IVM) paralyses L1 larvae soon after hatching, however, much lower concentrations (approximately 1 nM) are sufficient to inhibit development to the L3 stage which suggests that effects of the drug other than those relating to gross motor activity are responsible for the latter effect. The larval stages of IVM-susceptible H. contortus isolates from both Australia and South Africa, including isolates known to be resistant to levamisole or rafoxanide and/or the benzimidazoles, were equally sensitive to inhibition by AVMs. In contrast, 6 isolates of H. contortus resistant to IVM in vivo showed a reduced sensitivity to AVM inhibition of development. The order of potency of a limited range of AVMs as inhibitors of larval development was consistent with in vivo efficacy. Resistance ratios for IVM-resistant isolates were dependent on AVM structure, with AVM B2 the most sensitive probe for IVM resistance in the isolates tested.

Animals↗

Comparative brain ontogeny of the crayfish and clawed lobster: implications of direct and larval development.

The freshwater crayfish Cherax destructor and the lobster Homarus americanus have many similarities including life style, body form, and neural organization. However, the ontogenic history is very different in the two species. The development of Cherax is short and direct whereas the development of Homarus comprises three pelagic larval stages and takes more than twice as long from extrusion to benthic stages at constant temperature. In order to determine the progression of maturation of the nervous system in each species and the potential implications of pelagic forms on brain structure, the timing of appearance of 22 general and neural developmental events clearly identifiable in both species was compared. The onset of serotonin antigenicity in the different parts of the brain was chosen as one marker of neural development. During the first month of embryogenesis the timing of morphological, physiological, and neural events is similar in the two species. Morphological development is then accelerated in the crayfish near hatching time and over the two postembryonic stages before the advent of the independent benthic stage. Such heterochronic processes can at least partly account for the different developmental patterns in the two decapods. Among the characters showing similar timing in the two species is the formation of glomeruli (presumptive zones of synaptic contact) in the olfactory lobes of the deutocerebrum, although this event is embryonic in Homarus but postembryonic in Cherax. In contrast, glomerular formation in the accessory lobes is heterochronic: in both species, the glomeruli of the accessory lobes are acquired postembryonically, that is, 3 to 4 months earlier in Cherax than in Homarus. These data suggest that the development of the glomeruli in the olfactory lobes may depend primarily on internal developmental signals, whereas the triggering of glomerular formation in the accessory lobes may depend on external cues. The fact that, in Homarus, only the postlarval stages show mature accessory glomeruli may be a reflection of the functional requirements of benthic life.

Animals↗

Embryonic and larval development of the Drosophila mushroom bodies: concentric layer subdivisions and the role of fasciclin II.

Mushroom bodies (MBs) are the centers for olfactory associative learning and elementary cognitive functions in the arthropod brain. In order to understand the cellular and genetic processes that control the early development of MBs, we have performed high-resolution neuroanatomical studies of the embryonic and post-embryonic development of the Drosophila MBs. In the mid to late embryonic stages, the pioneer MB tracts extend along Fasciclin II (FAS II)-expressing cells to form the primordia for the peduncle and the medial lobe. As development proceeds, the axonal projections of the larval MBs are organized in layers surrounding a characteristic core, which harbors bundles of actin filaments. Mosaic analyses reveal sequential generation of the MB layers, in which newly produced Kenyon cells project into the core to shift to more distal layers as they undergo further differentiation. Whereas the initial extension of the embryonic MB tracts is intact, loss-of-function mutations of fas II causes abnormal formation of the larval lobes. Mosaic studies demonstrate that FAS II is intrinsically required for the formation of the coherent organization of the internal MB fascicles. Furthermore, we show that ectopic expression of FAS II in the developing MBs results in severe lobe defects, in which internal layers also are disrupted. These results uncover unexpected internal complexity of the larval MBs and demonstrate unique aspects of neural generation and axonal sorting processes during the development of the complex brain centers in the fruit fly brain.

Animals↗

Growth-blocking peptide titer during larval development of parasitized and cold-stressed armyworm.

Growth-blocking peptide (GBP) has been isolated for the first time from the haemolymph of the host armyworm Pseudaletia separata whose development was halted in the last larval instar stage by parasitization with the parasitoid wasp Cotesia kariyai. Recent studies demonstrated that GBP not only exists in the plasma (haemolymph without cells) of parasitized last instar larvae, but also in the plasma of nonparasitized penultimate (5th) instar larvae. Monoclonal antibodies were prepared to measure the titers of GBP in nonparasitized and parasitized larval plasma. One of three monoclonal antibodies raised against GBP, which is the most specific for GBP, was used to quantify the concentration of plasma GBP. As this antibody recognized two plasma peptides other than GBP in crude plasma fractions, each plasma peptide fraction was separated by a reversed phase HPLC, and then plasma GBP level was measured by ELISA. The highest level of plasma GBP detected on Day 0 of the penultimate instar larvae was gradually decreased throughout the larval growth except for the temporary increase on Day 0 of last larval instar. After parasitization on Day 0 of last larval instar, two peaks of plasma GBP titer were detected during the last larval instar, one day and six days after parasitization. This characteristic increase and decrease in plasma GBP level was also observed by transferring last instar larvae of the armyworm from 25 to 10 degrees C, as a result of which larvae delayed pupation by more than 15 days. From these results, it is reasonable to propose that plasma GBP in lepidopteran larvae might control certain upstream steps in a cascade of events leading to pupation; thus, an elevated level of plasma GBP interferes with normal metamorphosis from larvae to pupae.

Amino Acid Sequence↗

Effect of Campoletis sonorensis ichnovirus cys-motif proteins on Heliothis virescens larval development.

Polydnaviruses are obligate symbionts of some parasitic hymenopteran wasps responsible for modifying the physiology of their host lepidopteran larvae to benefit the endoparasite. Injection of Campoletis sonorensis ichnovirus (CsIV) into Heliothis virescens larvae alters larval growth, development and immunity but genes responsible for alterations of host physiology are not well described. Recent studies of polydnavirus genomes establish that these genomes encode families of related genes expressed in parasitized larvae. Here we evaluate five members of the CsIV cys-motif gene family for their ability to inhibit growth and development of lepidopteran larvae. To study the function of cys-motif proteins, recombinant proteins were produced from baculovirus expression vectors and injected or fed to H. virescens larvae in diet. rVHv1.1 was identified as the most potent protein tested causing a significant reduction in growth of H. virescens and Spodoptera exigua larvae. H. virescens larvae ingesting this protein also exhibited delayed development, reductions in pupation and increased mortality. Increased mortality was associated with chronic sub-lethal baculovirus infections. Taken together, these data indicate that the cys-motif proteins have pleiotropic effects on lepidopteran physiology affecting both development and immunity.

Amino Acid Motifs↗

In vitro effects of ACTH on interrenal corticosteroidogenesis during early larval development in rainbow trout.

Interrenal tissue from embryonic and larval rainbow trout (Oncorhynchus mykiss) was cultured in vitro and exposed to various doses of adrenocorticotropin (ACTH) to document the ontogeny of interrenal responsiveness to tropic stimulation. Resting and acute stress-induced changes in whole-body cortisol levels in vivo were also measured to determine if the corticosteroid stress response first develops with the onset of interrenal responsiveness to ACTH. No evidence was found that the hypothalamic-pituitary-interrenal (HPI) axis of rainbow trout is transiently activated prior to hatching. In vivo, a corticosteroid stress response was first observed 2 weeks after hatching, and stress-induced cortisol levels (at 1 hr poststress) were significantly higher 3 weeks after hatching than they were at 2 or 4 weeks after hatching. In contrast, cultured interrenal tissue produced significant levels of cortisol in response to ACTH at the time of hatching, and in vitro cortisol production by the interrenal increased significantly between 3 and 4 weeks after hatching. Interrenal sensitivity to ACTH did not change appreciably with development. We conclude that (1) the final maturation of the corticosteroid stress response in rainbow trout occurs at the level of the brain and/or sensory inputs and not at the level of the interrenal cell; (2) negative feedback mechanisms within the HPI axis develop 3 to 4 weeks after hatching; and (3) the period between 3 and 4 weeks after hatching may be homologous to the stress hyporesponsive period after birth in mammals and thus could be a stage when environmental influences can permanently alter the development of the corticosteroid stress response in rainbow trout.

Adrenocorticotropic Hormone↗

Density-dependent processes in the transmission of human onchocerciasis: relationship between the numbers of microfilariae ingested and successful larval development in the simuliid vector.

A previous paper reported that the intake of Onchocerca volvulus microfilariae (mff) by different species of Simulium is essentially proportional to the parasite load in the skin of infected carriers. This paper examines the fate of the ingested mff in susceptible vectors to assess the relationship between parasite intake and infective larval output in blackfly species with and without well-developed cibarial armatures. Analysis is based on data from 3 onchocerciasis endemic areas: Guatemala (S. ochraceum s.l.), West Africa (S. damnosum s.l./S. sirbanum) and the Amazonian focus between South Venezuela and Northern Brazil (S. guianense and S. oyapockense s.l.). The data, which include published and unedited information collected in the field, record experimental studies of parasite uptake by wild flies maintained in captivity until the completion of the extrinsic incubation period. The relationship between L3 output (measured as the mean number of successful larvae/fly or, as the proportion of flies with infective larvae) and average microfilarial intake, was strongly non-linear. This non-linearity was best represented by a sigmoid function in case of armed simuliids (S. ochraceum s.l., S. oyapockense s.l.), or by a hyperbolic expression in that of unarmed flies (S. damnosum s.l., S. guianense). These results are compatible, respectively, with the patterns of 'initial facilitation' and 'limitation' described in culicid vectors of lymphatic filariases. A maximum mean number of 1-3 L3/fly was observed in all 4 vectors. It is concluded that O. volvulus larval development to the infective stage is regulated by density-dependent mechanisms acting at the early phase of microfilarial migration out of the blackfly's bloodmeal. Damage by the bucco-pharyngeal armature may also be density dependent. A hypothesis, based on this density dependence is forwarded to explain initial facilitation, so far only recorded in vectors with well-developed cibarial teeth. Our results provide quantitative support for the conjecture that chemotherapy alone is likely to have a greater impact on reducing onchocerciasis transmission in endemic areas where the main vector has a toothed fore-gut than in foci where the vectors have unarmed cibaria.

Africa, Western↗

Nicotinamide in relation to dietary nicotinic acid and nine other vitamins and larval development of Ceratitis capitata (Diptera: Tephritidae).

This study was undertaken to understand why Ceratitis capitata larvae reared on a diet fortified with nine vitamins except nicotinic acid had 100% mortality, while those reared on a 10-vitamin-free diet had 66% survival (Chang, C. L.; Li, Q. X. Ann. Entomol. Soc. Am. 2004, 97, 536-540). Our results showed that nicotinamide was detected at a level of 0.07 microg/g in second-instar larvae reared on the 10-vitamin-free diet and 0.30 microg/g in the corresponding spent diet, while it was not detected in either the larvae reared on the diet fortified with 707 microg/g of nine vitamins (nicotinic acid absent) or the corresponding spent diet. Nicotinamide was detected at concentrations of 0.13 and 0.15 microg/g in the larvae fed the diets that were fortified with 707 microg/g of nine other vitamins and 2 and 20 microg/g of nicotinic acid, respectively, but it was not found in the larvae fed the 0.2 microg/g of nicotinic acid diet. Nicotinamide was detected at concentrations of 0.44, 0.52, and 0.55 microg/g in the spent diets that were fortified with the nine vitamins (707 microg/g) and 0.2, 2, and 20 microg/g of nicotinic acid, respectively. Nicotinamide adenine dinucleotide (NAD) was in the live larvae, but not in the dead larvae. These findings indicate that dietary nicotinic acid is converted to nicotinamide, which, in turn, is used to synthesize NAD, and suggest a positive relationship between C. capitata larvae survival rates and concentrations of dietary nicotinic acid and nitcotinamide in the larvae as well as in the spent diets. The result shows that nicotinamide derived from supplemental nicotinic acid is essential for the development and survival of C. capitata larvae. Nicotinamide may be a biomarker for larval survival and development.

Animals↗

Metamorphosis and early larval development of the flatfishes (Pleuronectiformes): an osmoregulatory perspective.

Flatfish (Pleuronectiformes) distribution in the environment is influenced by salinity, and varies among species and with developmental stage. Osmoregulatory ability likely plays an important role in defining species and developmental stage-specific distribution. Although the mechanisms of osmoregulation in adult and juvenile teleosts have been widely addressed, far less is known about their larval osmoregulatory physiology. Much of our current understanding of larval fish ion-regulation stems from studies using flatfishes, and this article reviews advances in this field, primarily from the point of view of the developing flatfishes. Addressed here are the ontogeny of salinity tolerance, the development of several important osmoregulatory tissues (the skin, gut, and gill), and the influence of the endocrine system on osmoregulation during early larval development and metamorphosis.

Animals↗

[Larval development of Paricterotaenia porosa (Cestoda: Cyclophyllidea, Dilepididae) in dipterans of the genus Chironomus, experimental hosts. On comparative larval forms of dilepidid flatworms (author's transl)].

The authors describe the larval evolution of Paricterotaenia porosa (Rud., 1810) Fuhrmann (1932) (Cestoda: Cyclophyllidea, Dilepididae) in three species belonging to the genus Chironomus (Diptera, Nematocera) used as experimental hosts. The protoscolex of the infestant larva is retracted into a cystic vesicule, the cysticercoïd is surrounded by a fragmented cercomer and a capsule. P. porosa in contrary to P. paradoxa (Rud., 1802) do not present asexual reproduction in its intermediate host. P. porosa and Anomotaenia constricta (Molin, 1858) Cohn (1900) show a similar evolution, though the cercomer appears sooner in the former. These cysticercoïds are regarded as Monocercus Villot (1883) the characteristics of which are given. The Monocercus is specific to the Dilepididae. The other larval types described in this family i.e. Cercoscolex and Plerocercus (Jackera, 1970) Plerocercoïd (Freeman, 1973) ans Strobilo cysticercoïd (Freeman, 1973) are compared to the larva of other families: Proteocephalidae, Mesocestoididae and Amabiliidae respectively.

Animals↗

The effect of thymectomy at different stages of larval development on the immune response of the clawed toad to sheep erythrocytes.

The role of the thymus during the first three weeks of larval life in the maturation of humoral immunity in the amphibian, Xenopus laevis, is examined. Thymectomy throughout this period dramatically affects the heterologous red cell response of young adults. Sheep erythrocyte (SRBC) administration elicited haemolytic antibody production in the blood and in the spleen (measured by the appearance of plaque forming cells) of control animals, but failed to do so in all thymectomized toadlets. Moreover, use of the immunocyto-adherence assay, which proved to be a sensitive test for SRBC reactivity in control Xenopus, indicated a complete absence of induced responsiveness to this antigen in the spleens of thymectomized toadlets, even in those animals thymectomized as late as 22 days of age. In contrast to allograft immunity in Xenopus, Which is suppressed only by thymic ablation during the first two weeks of larval life, the development of reactivity to heterologous erythrocytes therefore requires an additional period of thymic influence during ontogeny. These experiments support a concept of thymus-dependent cell heterogeneity in amphibians.

Animals↗

In vitro demonstration of in utero larval development in an oviparous parasitic nematode: Haemonchus contortus.

The life-cycle of Haemonchus contortus, a pathogenic stomach nematode of sheep, is typical of those of the other members of the superfamily Trichostrongyloides, all of which require a period of development outside the definitive host. Classically, gravid H. contortus, known as strictly oviparous, releases her eggs into the abomasal lumen. The eggs are passed out in the faeces in which they hatch into the 1st-stage larvae and then develop into the infective 3rd-stage larvae. We have developed a method to study the fate of eggs within gravid worms. Using this procedure, we have shown that H. contortus may also exhibit viviparity with apparently normal development from the egg to the 4th larval stage taking place within the gravid female maintained in vitro. On the basis of these observations we speculate that viviparity might occur in vivo with consequent autoinfections; if so, this might explain some puzzling clinical and epidemiological features of haemonchosis, as well as the incomplete efficacy of current control measures.

Abomasum↗