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Independent temporal expression of two N-substituted aminoacyl-tRNA hydrolases during the development of Artemia salina.

Encysted embryos of the crustacean Artemia salina contain an enzymatic activity which hydrolyzes N-acetylphenylalanyl-tRNA to N-acetylphenylalanine and tRNA. The enzyme apparently does not hydrolyze other free or N-substituted aminoacyl-tRNAs. The levels of this enzyme do not significantly change during embryonic and early larval development. In contrast, an unspecific hydrolase active on several N-substituted aminoacyl-tRNAs is practically absent in the encysted embryos and during embryogenesis and appears abruptly during larval development. The independent temporal expression of these two hydrolases during Artemia salina differentiation makes this organism siuitable for the study of the physiological role of these enzymes.

Animals↗

Krüppel expression during postembryonic development of Drosophila.

The Drosophila gap gene Krüppel (Kr) codes for a Zn2+ finger transcription factor important in embryo patterning, segment identity, and specification of various cell types. Based on Northern blot analysis, Kr function has been thought to be restricted to embryogenesis. I report here that Kr is also expressed during larval development, primarily in the fat body. Using both a P-element enhancer-trap line and a portion of the Kr promoter fused to lacZ, I find the activation of Kr in the fat body does not begin until the early-third-instar stage. Earlier in larval development, Kr is detected in the nerve cells associated with the gut. Kr's role as a transcription factor during early embryogenesis is well established. After gastrulation Kr protein is detected in a variety of cell types and is probably involved in cell specification. Kr is not detected in fat cells during embryogenesis nor during the first- and second-instar stages. The delayed expression of Kr in the fat cells occurs at a time in which the larva is undergoing a reprogramming in preparation for metamorphosis. It is possible that Kr serves as a transcriptional regulator in the fat body in this last larval instar. During this last larval stage, several fat-body-specific genes are transcriptionally activated in response to increasing levels of ecdysone. The possible role of Kr in the expression of these ecdysone-regulated genes is discussed.

Animals↗

Biometrical analysis of larval digging behavior in Drosophila melanogaster.

Digging behavior of D. melanogaster larvae increases as larval development proceeds. Diallel crosses were made to analyze genetically digging behavior at 72 and 108 h of larval age. Additive and dominance variation was found, dominance being principally to dig. Dominance to dig is higher at 108 than 72 h of development; additivity does not substantially change between these two larval ages. At 72 h of larval age, depending on the cross, I found (i) dominance to dig, (ii) dominance to nondig, (iii) overdominance to dig, and (iv) no dominance to dig. At 108 h of larval development I detected (i) dominance to dig and (ii) overdominance to dig. Thus, diversity of response in the F1 was greater at 72 than 108 h of larval development. These age-related changes in larval digging behavior of D. melanogaster seem to reflect epigenetic changes in the patterns of gene expressions.

Animals↗

Gradual appearance of a regulated retinotectal projection pattern in Xenopus laevis.

The topographic projection pattern formed by the retinal ganglion cell axons in the tectum of the lower vertebrate appears to require positional cues that guide the optic nerve fibers to their appropriate targets. One approach to understanding these positional cues or "positional information" has been to investigate changes in the pattern of the retinotectal projection after surgical manipulation of the embryonic eyebud. Analysis of these apparent changes in the patterns of positional information in the eye, termed "pattern regulation," may provide clues to both the nature of positional information and the mechanisms by which it is assigned to cells in the eyebud. Here we examine pattern regulation in the Xenopus visual system following the replacement of the temporal half of a right eyebud with the temporal half of a left eyebud. This manipulation requires that the left half-eyebud be inverted along its dorsoventral axis. Electrophysiological maps of these compound eyes in postmetamorphic frogs reveal regulated maps; the cells in the temporal half of the NrTl eye project to the tectum with a dorsoventral polarity appropriate for their position in the host eye and not appropriate for the original positions of the grafted cells in the donor eyebud. Paradoxically, the regulated patterns are not apparent in the projections of the original grafted eyebud cells during early larval development. Using fiber-tracing and electrophysiological mapping techniques, we now show that the regulated patterns appear gradually in the projections made by peripheral retinal cells added during mid-larval development. Because the regulation occurs relatively late in development and probably only in the peripheral retinal cells, simple models of epimorphic or morphallactic regulation do not appear to fit this system. Thus, new or more complex models must be invoked to explain the phenomenon of pattern regulation in the developing visual system of Xenopus.

Animals↗

Ontogeny of regulation of gill and lung ventilation in the bullfrog, Rana catesbeiana.

Gill and lung ventilatory frequencies at 20-23 degrees C were recorded in five different larval stages and in the adults of the bullfrog, Rana catesbeiana (n = 101, body mass 40 mg-90 g). Ventilatory frequencies in unanesthetized, unrestrained animals were determined (1) after normoxic acclimation, (2) during acute hypoxic and hyperoxic exposure, and (3) after brief but intense activity. Gill ventilation frequency (fG) under normoxic, resting conditions was 110-120 cycles X min-1 immediately after hatching, but fell to and remained at 40-50 cycles X min-1 for the remainder of larval development. Activity caused a sharp decrease in fG in newly hatched larvae, a sharp increase in larvae between stages IV-XIV, and no change in fG in all older larval states. Hypoxia increased fG in younger larvae up to developmental stage XIV, but had no effect upon fG in older larvae. Lung ventilation was rare in normoxic, resting larvae up to stage X. Thereafter until metamorphosis lung ventilation frequency (fL) was 2-6 breaths X h-1, with fL in adults being much higher at 1-3 breaths X min-1. Activity did not affect fL in any larval stage, but markedly increased fL in adults. Hypoxia had no significant effect on mean fL in larvae below stage XX. Mean values of fL increased during acute hypoxic exposure in most adults, but these changes were not significant. Collectively, these data indicate that progressive larval development is accompanied by a decline in reflex regulation of branchial ventilation frequency well before reabsorption of gills occurred. At the same time, respiratory responses are 'transferred' to the lung prior to metamorphosis and the attendant increasing dependence on air breathing.

Animals↗

Caterpillar dermatitis in two siblings due to the larvae of Thaumetopoea processionea L, the oak processionary caterpillar.

Two siblings, 16 months and 5 years of age, came into contact with the urticating hairs (setae) of oak processionary caterpillars, the larvae of Thaumetopoea processionea L., when the family moved to a suburb of Vienna, where mass gradation of T. processionea had started the year before. The setae were being spread by the wind from an infested oak tree in the neighbourhood. Both children repeatedly suffered bouts of dermatitis during the 10 weeks of the larval development. Owing to the fact that T. processionea often infests oak trees, whether isolated or at the edges of forests, there is a high likelihood of people being affected. Children frequently explore their surroundings and are at an even greater risk of developing lepidopterism. Caterpillar dermatitis should therefore be taken into consideration in the differential diagnosis of a pruritic rash in infants from regions with caterpillar-infested trees, especially during the larval development of T. processionea .

Air Pollutants↗

An essential role in molting and morphogenesis of Caenorhabditis elegans for ACN-1, a novel member of the angiotensin-converting enzyme family that lacks a metallopeptidase active site.

Genome sequence analyses predict many proteins that are structurally related to proteases but lack catalytic residues, thus making functional assignment difficult. We show that one of these proteins (ACN-1), a unique multi-domain angiotensin-converting enzyme (ACE)-like protein from Caenorhabditis elegans, is essential for larval development and adult morphogenesis. Green fluorescent protein-tagged ACN-1 is expressed in hypodermal cells, the developing vulva, and the ray papillae of the male tail. The hypodermal expression of acn-1 appears to be controlled by nhr-23 and nhr-25, two nuclear hormone receptors known to regulate molting in C. elegans. acn-1(RNAi) causes arrest of larval development because of a molting defect, a protruding vulva in adult hermaphrodites, severely disrupted alae, and an incomplete seam syncytium. Adult males also have multiple tail defects. The failure of the larval seam cells to undergo normal cell fusion is the likely reason for the severe disruption of the adult alae. We propose that alteration of the ancestral ACE during evolution, by loss of the metallopeptidase active site and the addition of new protein modules, has provided opportunities for novel molecular interactions important for post-embryonic development in nematodes.

Amino Acid Sequence↗

Further studies on the development and availability of infective larvae of bovine gastrointestinal trichostrongylids on pasture in eastern Nigeria.

The dynamics of pasture populations of infective larvae (L3) of Cooperia, Haemonchus and Trichostrongylus species were studied at Nsukka, eastern Nigeria, during April to November 1984. Six paddocks were contaminated artificially and one was contaminated naturally. Five of the paddocks, P1-P5, were sequentially contaminated with faeces of naturally infected cattle at approximately 4-6-weekly intervals. Paddocks P6 and P7 were repeatedly contaminated every 4-6 weeks artificially and by the naturally infected cattle, respectively. Larval development and survival occurred very readily during the wet season (April-October) but apparently ceased in November at the start of the dry season. Larval migration, however, occurred not only during the rains but also during the first 4 weeks of the dry season. Single contaminations during the rains quickly gave rise to single waves of infestation which also declined rapidly, in spite of the continuously favourable conditions for larval development and survival. The repeated contaminations produced three and four distinct and relatively short-lived larval peaks, respectively, with the first three peaks on both paddocks, namely the May, July and September/October peaks, being coincident. The four waves of herbage infestation on P7, which occurred at approximately 4-5 weekly intervals, were considered to have originated from four separate generations of the three trichostrongylids. However, Trichostrongylus sp. predominated in the first (May) peak while Cooperia and Haemonchus dominated the later peaks.

Animal Feed↗

Development of sibling inbred sea urchins: normal embryogenesis, but frequent postembryonic malformation, arrest and lethality.

Inbred lines of Strongylocentrotus purpuratus descended from a single pair of wild animals were constructed by sibling mating. We describe results from a systematic series of crosses in which eggs from F2 and from F3 females were fertilized respectively with sperm from their sibling males. Observations were also made on self-fertilized cultures derived from several naturally occurring hermaphrodites. Morphological development, survival efficiency, and expression of three territorial embryonic markers were assayed in the embryos developing from these crosses. Unexpectedly, out of > 90 controlled crosses, we observed no developmental failures whatsoever, up to the end of embryogenesis (i.e., onset of feeding) that could be attributed to homozygous, zygotically acting recessive genes. However, during postembryonic larval development, lethality, morphological malformation, and arrest are observed in inbred cultures at a high frequency. The incidence of these zygotic developmental failures is such that it appears that there is at least one recessive genetic defect affecting larval development per haploid parental genome. The relative imperviousness of the basic embryonic process to defects arising from homozygosity is consistent with other evidence implying that territorial specification in sea urchin embryogenesis is controlled by maternally rather than zygotically expressed gene products.

Animals↗

Ovoviviparity in platyhelminth life-cycles.

The encapsulated embryos of platyhelminths may be retained and complete their development in utero in a range of circumstances. However, hatching within the parent (the criterion of ovoviviparity) is relatively rare and larvae generally emerge only after deposition. Viviparity is characterized by the nutritional dependency of the unencapsulated larva upon the parent, but in several cases larvae retained within a shell also receive parental nutrients during intra-uterine development. Uptake of exogenous nutrients via shell pores occurs in Schistosoma mansoni but the eggs, which gain all the advantages of intra-uterine retention, are supported by host nutrients. Intra-uterine larval development avoids the hazards of development in the external environment and eliminates the time delay between oviposition and infection. Deposition of immediately infective offspring may be concentrated in time and space to exploit periods of host vulnerability. The control and precision of transmission is illustrated by examples in which the opportunity for invasion is restricted because of either host behaviour or environmental instability. This strategy has been an important factor in the evolution of polystomatid monogeneans, and its effectiveness is demonstrated by comparison of the life-cycles of Polystoma integerrimum and Pseudodiplorchis americanus. Ovoviviparity also increases reproductive potential in some polystomatids by extending the period of multiplication and by increasing established populations through internal re-infection. In Eupolystoma alluaudi, the capacity for ovoviviparity is programmed into larval development and this regulates population growth within individual hosts.

Adaptation, Physiological↗

Expression of a midgut-specific cadherin BT-R1 during the development of Manduca sexta larva.

The btr-1 gene of Manduca sexta (GenBank AF319973) encodes a cadherin, BT-R(1) (210-kDa), which contains 12 ectodomain modules in association with a number of motifs potentially involved in interactions with cadherin and integrin. The molecule is a target receptor for Bacillus thuringiensis Cry1A toxins that bind to BT-R(1) with high affinity and specificity. BT-R(1) is localized exclusively in the midgut epithelium. The amount of BT-R(1) protein increases dramatically during larval development, paralleling accumulation of its mRNA. The 5'-UTR of the btr-1 gene contains sequence motifs that most likely recruit specific transcription factors, particularly, those that determine posterior patterning and that control intestinal cell proliferation, differentiation and identity during development. The increase in abundance of BT-R(1) may be required to support not only the differentiation of the epithelial cells but also the establishment of physiological function and structural integrity of the midgut during larval development in M. sexta. We believe that BT-R(1) is essential to larval midgut epithelial organization during rapid cell proliferation and tissue growth in M. sexta because disruption of such organization and functionality occasioned by the binding of the Cry1A toxins of B. thuringiensis to BT-R(1) causes death to the insect.

Animals↗

[Change in intensity of respiration in development of some invertebrates].

We studied the dynamic of respiration intensity during ontogenesis of flat worms (Dugesia tigrina), molluscs (Anodonta piscinalis and Viviparus viviparus, and insects (Leptinotarsa decemlineata). In planarians that reproduce vegetatively, the intensity of respiration increases just after fission and decreases at the subsequent phases of growth. In A. piscinalis, this index of metabolism increases during embryonic and early larval development and decreases at the later developmental stages. In V. viviparus, which develops in the female genital tract, the intensity of respiration remains unchanged during embryogenesis and decreases during late embryogenesis and subsequent phases of growth. In L. decemlineata, the intensity of respiration increases during embryonic and early larval development and then decreases to undergo cyclic changes times to molts. This index markedly decreases in the pupae, increases in the beginning of imaginal period, and then again decreases.

Animals↗

Thyroid and pituitary gland development from hatching through metamorphosis of a teleost flatfish, the Atlantic halibut.

Fish larval development, not least the spectacular process of flatfish metamorphosis, appears to be under complex endocrine control, many aspects of which are still not fully elucidated. In order to obtain data on the functional development of two major endocrine glands, the pituitary and the thyroid, during flatfish metamorphosis, histology, immunohistochemistry and in situ hybridization techniques were applied on larvae of the Atlantic halibut (Hippoglossus hippoglossus), a large, marine flatfish species, from hatching through metamorphosis. The material was obtained from a commercial hatchery. Larval age is defined as day-degrees (D degrees =accumulated daily temperature from hatching). Sporadic thyroid follicles are first detected in larvae at 142 D degrees (27 days post-hatch), prior to the completion of yolk sack absorption. Both the number and activity of the follicles increase markedly after yolk sack absorption and continue to do so during subsequent development. The larval triiodothyronine (T(3)) and thyroxine (T(4)) content increases, subsequent to yolk absorption, and coincides with the proliferation of thyroid follicles. A second increase of both T(3) and T(4) occurs around the start of metamorphosis and the T(3) content further increases at the metamorphic climax. Overall, the T(3) content is lower than T(4). The pituitary gland can first be distinguished as a separate organ at the yolk sack stage. During subsequent development, the gland becomes more elongated and differentiates into neurohypophysis (NH), pars distalis (PD) and pars intermedia (PI). The first sporadic endocrine pituitary cells are observed at the yolk sack stage, somatotrophs (growth hormone producing cells) and somatolactotrophs (somatolactin producing cells) are first observed at 121 D degrees (23 days post-hatch), and lactotrophs (prolactin producing cells) at 134 D degrees (25 days post-hatch). Scarce thyrotrophs are evident after detection of the first thyroid follicles (142 D degrees ), but coincident with a phase in which follicle number and activity increase (260 D degrees ). The somatotrophs are clustered in the medium ventral region of the PD, lactotrophs in the anterior part of the PD and somatolactotrophs are scattered in the mid and posterior region of the pituitary. At around 600 D degrees , coinciding with the start of metamorphosis, somatolactotrophs are restricted to the interdigitating tissue of the NH. During larval development, the pituitary endocrine cells become more numerous. The present data on thyroid development support the notion that thyroid hormones may play a significant role in Atlantic halibut metamorphosis. The time of appearance and the subsequent proliferation of pituitary somatotrophs, lactotrophs, somatolactotrophs and thyrotrophs indicate at which stages of larval development and metamorphosis these endocrine cells may start to play active regulatory roles.

Animals↗

Biotic and abiotic parameters associated with an epizootic of Coelomomyces punctatus in a larval population of the mosquito Anopheles quadrimaculatus.

Biotic and abiotic parameters associated with an epizootic of the fungus Coelomomyces punctatus in larval populations of the mosquito Anopheles quadrimaculatus were investigated for three mosquito breeding seasons (1986-1988) in two adjacent farm ponds in North Carolina. In the first pond, the prevalence of infected larvae averaged 42% (range 0-85%) for collections made weekly from May 1 to November 20, 1986, but larvae did not occur in this pond in 1987. Infection rates in the adjacent pond, sampled during the mosquito breeding seasons of 1987 and 1988, declined from 10.9% (range 0-27.5%) in 1987 to 2.5% (range 0-14.2%) in 1988. Correlation analyses between the number of female copepods and fungal infection rates in sentinel mosquitoes were significant (P < 0.01) for Acanthocyclops robustus but insignificant for eight other species. Infections obtained in sentinel larvae placed in the ponds for 3 hr intervals indicated that C. punctatus infected larvae around sundown. Infection rates for field-collected larvae increased with the stage of larval development. However, experiments with sentinel larvae showed that early instars were more susceptible to infection than later instars, suggesting that the higher infection rates in late instars resulted from individual larvae being infected by two or more zygotes during larval development. Standard multiple regression analyses, used to determine the relationship between seasonal infection rates and water chemistry, weather variables, and the abundance of early and late instar larvae, showed that the abundance of late instars was the only independent variable common to linear models. The models only accounted for 20 and 9% of the variation in larval infection rates for 1987 and 1988, respectively. These results indicate that of the parameters examined, the seasonal abundance of the copepod, A. robustus, was the most important factor (or variable) correlated with the prevalence of mosquito infection.

Animals↗

Mating and reproduction of predaceous diving beetles, Dytiscus sharpi, observed under artificial breeding conditions.

Mating season and embryonic development of the predaceous diving beetles, Dytiscus sharpi, (Coleoptera; Dytiscidae) were observed under artificial breeding conditions. Female and male adult insects started mating from November to March and gave first instar larvae mainly in April. When the mating was artificially delayed until February, first instar larvae appeared from the end of March to the middle of May. I also investigated the effects of temperature on larval development. Apparent hatchability of eggs was not affected by high temperature, however, their normal development after hatching was significantly interfered. Most of the first instar larvae kept at 20-25 degrees C from before hatching died within one day after hatching. By contrast, juveniles kept outdoors (7.0-20.9 degrees C) could develop at least until second instar larvae. Temperature >23 degrees C after hatching had no effects on larval development. From these observations, it was concluded that the reproduction strategy of Dytiscus sharpi, i.e. mating in late autumn and hatching in early spring would be the reasonable results of adaptation to the warm habitats where they are collected.

Animals↗

Ecdysteroid receptors in the central nervous system of Manduca sexta: their changes in distribution and quantity during larval-pupal development.

Ecdysteroids act initially by binding to nuclear and possibly also extranuclear receptors. The presence and expression of these receptors in the insect brain was investigated in the present study as a means of defining these neurons involved in ecdysteroid-regulated processes at different developmental stages. Early in the fifth larval stadium of Manduca sexta, when endogenous ecdysteroid levels are low, receptors for ecdysteroids in cerebral neurons are either absent or present at low levels. Receptors can be reliably detected only on day 0 and are not found again until day 3.5, at the beginning of the commitment peak in the ecdysteroid titer, when they occur in a small stage-specific population of cells. At this time, ecdysteroid receptors are found mainly in nuclei but are also observed at low levels in cytoplasm. By day 4.8, ecdysteroid receptors are exclusively nuclear, and the number of target cells has increased dramatically in several brain regions, including those with known neurosecretory cell groups. This population and organization of ecdysteroid target cells is constant up to day 6, after which time the number of target neurons declines. By day 7.8, only 10% of the number of labelled neurons seen on days 4.8-6.8 remain in peripheral areas. In the pupal brains, ecdysteroid receptors reappear in a new population of neurons. The results indicate changes in the genomic regulation of a varying neuron population by ecdysteroids during fifth stadium development.

Animals↗

Developmental changes in oxygen consumption regulation in larvae of the South African clawed frog Xenopus laevis

Well-developed larval Xenopus laevis (NF stages 58&shy;66) are oxygen regulators, at least during mild hypoxia. When and how they change from oxygen conformers (the presumed condition of the fertilized egg) to oxygen regulators is unknown. Also unknown is how anaerobic metabolic capabilities change during development, especially in response to acute hypoxia, and to what extent, if any, anaerobiosis is used to supplement aerobic metabolism. Consequently, we have investigated resting rates of oxygen consumption (M.O2) and concentrations of whole-body lactate (lactic acid) during development in normoxia and in response to acute hypoxia in Xenopus laevis. M.O2 increased in an episodic, non-linear fashion during development. Resting, normoxic M.O2 increased about tenfold (to approximately 0.20 &micro;mol g-1 h-1) between NF stages 1&shy;39 and 40&shy;44, and then another tenfold between NF stages 45&shy;48 and 49&shy;51 (to approximately 2.0 &micro;mol g-1 h-1), remaining at about 2 &micro;mol g-1 h-1 for the remainder of larval development. M.O2 reached its highest level in newly metamorphosed frogs (nearly 4 &micro;mol g-1 h-1), before decreasing to about 1.0 &micro;mol g-1 h-1 in large adults. X. laevis embryos and larvae up to NF stage 54&shy;57 were oxygen conformers when exposed to variable levels of acute hypoxia. The only exception was NF stage 45&shy;48 (external gills present yet body mass still very small), which showed some capability of oxygen regulation. All larvae older than stage 54&shy;57 and adults were oxygen regulators and had the lowest values of Pcrit (the oxygen partial pressure at which M.O2 begins to decline). Whole-body lactate concentration in normoxia was about 1 &micro;mol g-1 for all larval groups, rising to about 12 &micro;mol g-1 in adults. Concentrations of lactic acid in NF stages 1&shy;51 were unaffected by even severe ambient hypoxia. However, whole-body lactate levels in NF stages 52&shy;66 increased in response to severe hypoxia, indicating that some anaerobic metabolism was being used to supplement diminishing aerobic metabolism. The largest increases in concentration of lactate occurred in late larvae and adults.

Journal Article↗

Conservation of Hox/ParaHox-related genes in the early development of a cnidarian.

To clarify the relationship between axial patterning in cnidarians and bilaterians, we have investigated the embryonic development of the hydrozoan Podocoryne carnea. The expression of Hox-like homeobox genes was analyzed by RT-PCR and in situ hybridization. Cnox1-Pc, an anterior Hox gene, is a maternal message. It is present throughout larval development, first weakly in all blastomeres and later restricted mostly to the anterior pole of the planula. Gsx, an anterior ParaHox gene, is first seen in the anterior endoderm but also extends into posterior regions. Cnox4-Pc, an orphan Hox-like gene, is expressed in the egg as a ring-shaped cloud around the germinal vesicle. After fertilization, the message remains in most animal blastomeres. When the embryo elongates in late blastula, staining is restricted to a few cells at the posterior pole where gastrulation will start. However, once gastrulation starts, the Cnox4-Pc signal disappears and is absent in later stages of larval development. Phylogenetic analysis shows that not all cnidarian Hox-like genes have recognizable orthologues in bilaterian groups. However, the expression analysis of Cnox1-Pc and Gsx correlates to some extent with the expression pattern of cognate genes of bilaterians, confirming the conservation of genes involved in organizing animal body plans and their putative common ancestral origin.

Amino Acid Sequence↗