Metamorphic acceleration following the exposure to lithium and selenium on American bullfrog tadpoles (Lithobates catesbeianus).

To regulate the presence of contaminants in Brazilian water, the Brazilian Environmental Council (CONAMA) promulgates regulations regarding the concentrations of given compounds that are supposed to be safe for aquatic life. Considering these regulations, this study tested the effects of considered safe levels of lithium (2.5 mgL-1) and selenium (0.01 mgL-1), isolated and mixed, on the American bullfrog (Lithobates catesbeianus) tadpoles. The evaluation was done through the use of biomarkers of larval development as total wet weight (TWW), snout-vent-length (SVL), hind-limb-length (HLL), activity level (AL), histologic evaluation of the thyroid gland and the mortality rate. The tadpoles were allocated into four groups (n = 20 each): a control group (CT); a group exposed to lithium (LI), a group exposed to selenium (SE), and a group exposed to both lithium and selenium (SELI). The whole assay was carried out over 21 days, with two rounds of data collection (on 7th and 21st day) to evaluate the responses over time. A statistical reduction in the AL was observed in the tadpoles from the LI and SELI groups after 7 days of exposure, the same pattern was observed after 21 days. Histological analyses of the thyroid gland showed signs of up-regulation (i.e. statistic reduction in number and area of the follicles, as well a significant reduction in the area of the gland) in all exposed groups, which represents an endocrine response as an adaptative strategy to deal with polluted aquatic environment. The stress triggered by the polluted medium is discussed.

To eat or not to eat: ontogeny of hypothalamic feeding controls and a role for leptin in modulating life-history transition in amphibian tadpoles.

Many animal life histories entail changing feeding ecology, but the molecular bases for these transitions are poorly understood. The amphibian tadpole is typically a growth and dispersal life-history stage. Tadpoles are primarily herbivorous, and they capitalize on growth opportunities to reach a minimum body size to initiate metamorphosis. During metamorphic climax, feeding declines, at which time the gastrointestinal (GI) tract remodels to accommodate the carnivorous diet of the adult frog. Here we show that anorexigenic hypothalamic feeding controls are absent in the tadpole, but develop during metamorphosis concurrent with the production of the satiety signal leptin. Before metamorphosis there is a large increase in leptin mRNA in fat tissue. Leptin receptor mRNA increased during metamorphosis in the preoptic area/hypothalamus, the key brain region involved with the control of food intake and metabolism. This corresponded with an increase in functional leptin receptor, as evidenced by induction of socs3 mRNA and phosphorylated STAT3 immunoreactivity, and suppression of feeding behaviour after injection of recombinant frog leptin. Furthermore, we found that immunoneutralization of leptin in tadpoles at metamorphic climax caused them to resume feeding. The absence of negative regulation of food intake in the tadpole allows the animal to maximize growth prior to metamorphosis. Maturation of leptin-responsive neural circuits suppresses feeding during metamorphosis to facilitate remodelling of the GI tract.

Evolution and development of scyphozoan jellyfish.

Scyphozoan jellyfish, or scyphomedusae, are conspicuous members of many ocean ecosystems, and have large impacts on human health and industry. Most scyphomedusae are the final stage in a complex life cycle that also includes two intermediate stages: the larval planula and benthic polyp. In species with all three life-cycle stages, the metamorphosis of a polyp into a juvenile scyphomedusa (ephyra) is termed strobilation, and polyps can produce one ephyra (termed monodisc strobilation) or many ephyrae (termed polydisc strobilation). In contrast to species with planula, polyp and medusa stages, a handful of scyphozoan species possess modified life cycles with reduced or absent stages. The evolutionary patterns associated with strobilation and life-cycle type have not been thoroughly investigated, and many studies of ephyra development and strobilation induction are not yet synthesized. Herein, I place the development of scyphomedusae in an evolutionary context. I first review the current evolutionary hypotheses for Scyphozoa. Next, I review what is known about scyphomedusa development across a broad diversity of species, including the first signs of strobilation, the formation of strobila segments, and the morphogenesis of ephyrae. I then review cases where the canonical scyphozoan life cycle has been modified, and take advantage of phylogenetic hypotheses to place these observations in an evolutionary context. I show that the evolution of monodisc strobilation occurred at least twice, and that the loss of intermediate life-cycle stages occurred several times independently; by contrast, the reduction of the medusa stage appears to have occurred within a single clade. I then briefly review the major natural cues of strobilation induction. Finally, I summarize what is currently known about the molecular mechanisms of strobilation induction and ephyra development. I conclude with suggestions for future directions in the field.

Lamprey metamorphosis: Thyroid hormone signaling in a basal vertebrate.

As one of the most basal living vertebrates, lampreys represent an excellent model system to study the evolution of thyroid hormone (TH) signaling. The lamprey hypothalamic-pituitary-thyroid and reproductive axes overlap functionally. Lampreys have 3 gonadotropin-releasing hormones and a single glycoprotein hormone from the hypothalamus and pituitary, respectively, that regulate both the reproductive and thyroid axes. TH synthesis in larval lampreys takes place in an endostyle that transforms into typical vertebrate thyroid tissue during metamorphosis; both the endostyle and follicular tissue have all the typical TH synthetic components found in other vertebrates. Furthermore, lampreys also have the vertebrate suite of peripheral regulators including TH distributor proteins (THDPs), deiodinases and TH receptors (TRs). Although at the molecular level the components of the lamprey thyroid system are ancestral to other vertebrates, their functions have been largely conserved. TH signaling as it relates to lamprey metamorphosis represents a particularly interesting phenomenon. Unlike other metamorphosing vertebrates, lamprey THs increase throughout the larval period, peak prior to metamorphosis and decline rapidly at the onset of metamorphosis; patterns of deiodinase activity are consistent with these increases and declines. Moreover, goitrogens (which suppress TH levels) initiate precocious metamorphosis, and exogenous TH treatment blocks goitrogen-induced metamorphosis and disrupts natural metamorphosis. Despite this clear physiological difference, TH action via TRs is consistent with higher vertebrates. Based on observations that TRs are upregulated in a tissue-specific fashion during morphogenesis and the finding that lamprey TRs upregulate genes via THs in a fashion similar to higher vertebrates, we propose the following hypothesis for further testing. THs have a dual role in lampreys where high TH levels promote larval feeding and growth and then at the onset of metamorphosis TH levels decrease rapidly; at this time the relatively low TH levels function via TRs in a fashion similar to that of other metamorphosing vertebrates.

A lamprey view on the origins of neuroendocrine regulation of the thyroid axis.

This mini review summarizes the current knowledge of the hypothalamic-pituitary-thyroid (HPT) endocrine system in lampreys, jawless vertebrates. Lampreys and hagfish are the only two extant members of the class of agnathans, the oldest lineage of vertebrates. The high conservation of the hypothalamic-pituitary-gonadal (HPG) axis in lampreys makes the lamprey model highly appropriate for comparative and evolutionary analyses. However, there are still many unknown questions concerning the hypothalamic-pituitary (HP) axis in its regulation of thyroid activities in lampreys. As an example, the hypothalamic and pituitary hormone(s) that regulate the HPT axis have not been confirmed and/or characterized. Similar to gnathostomes (jawed vertebrates), lampreys produce thyroxine (T4) and triiodothyronine (T3) from thyroid follicles that are suggested to be involved in larval development, metamorphosis, and reproduction. The existing data provide evidence of a primitive, overlapping yet functional HPG and HPT endocrine system in lamprey. We hypothesize that lampreys are in an evolutionary intermediate stage of hypothalamic-pituitary development, leading to the emergence of the highly specialized HPG and HPT endocrine axes in jawed vertebrates. Study of the ancient lineage of jawless vertebrates, the agnathans, is key to understanding the origins of the neuroendocrine system in vertebrates.

Detritus Quality and Locality Determines Survival and Mass, but Not Export, of Wood Frogs at Metamorphosis.

Single-site experiments have demonstrated detritus quality in wetlands can have strongly negative, neutral, and even positive influences on wildlife. However, an examination of the influence of detritus quality across several regions is lacking and can provide information on whether impacts from variation in detritus quality are consistent across species with wide ranges. To address this gap in regional studies we examined effects of emergent and allochthonous detritus of different nutrient qualities on amphibians and assessed a mechanism that may contribute to potential impacts. We used aquatic mesocosms to raise wood frogs (Rana sylvatica) from two regions of the United States with whole plants from purple loosestrife (Lythrum salicaria), leaf litter from native hardwood trees, and a mixture of both. We examined several metrics of amphibian fitness and life history, including survival, number of days to metamorphosis, and size at metamorphosis. Further, we quantified whether the effects of detritus type could translate to variation in anuran biomass or standing stock of nitrogen or phosphorus export. Our results show detritus with high nutrient quality (purple loosestrife) negatively influenced survival of wood frogs, but increased size of metamorphic individuals in two different regions of the United States. Despite the decrease in survival, the increase in size of post-metamorphic anurans raised with high quality detritus resulted in anuran biomass and standing stock of N and P export being similar across treatments at both locations. These results further demonstrate the role of plant quality in shaping wetland ecosystem dynamics, and represent the first demonstration that effects are consistent within species across ecoregional boundaries.

Life-history evolution: at the origins of metamorphosis.

Metamorphosis is a widespread life history strategy of animals but apart from some model organisms it is poorly characterized. A recent study of moon jellies highlights the similarities and differences between the various types of metamorphosis and illuminates its molecular determinants.

Regulation of polyp-to-jellyfish transition in Aurelia aurita.

BACKGROUND: The life cycle of scyphozoan cnidarians alternates between sessile asexual polyps and pelagic medusa. Transition from one life form to another is triggered by environmental signals, but the molecular cascades involved in the drastic morphological and physiological changes remain unknown. RESULTS: We show in the moon jelly Aurelia aurita that the molecular machinery controlling transition of the sessile polyp into a free-swimming jellyfish consists of two parts. One is conserved and relies on retinoic acid signaling. The second, novel part is based on secreted proteins that are strongly upregulated prior to metamorphosis in response to the seasonal temperature changes. One of these proteins functions as a temperature-sensitive "timer" and encodes the precursor of the strobilation hormone of Aurelia. CONCLUSIONS: Our findings uncover the molecule framework controlling the polyp-to-jellyfish transition in a basal metazoan and provide insights into the evolution of complex life cycles in the animal kingdom.

As forças formativas etéricas do ser humano e suas metamorfoses nas doenças autoimunes: Contribuição para uma fisiologia e fisiopatologia na biografia humana / The etheric driving forces of the human being and its metamorphosis in autoimmunes diseases: Contribution to the knowledge about physiology and physiopathology on the human biography

O organismo físico do ser humano é sustentado na sua configuração através do corpo etérico, o qual realiza isso, no nível da substância, por meio de sua atividade na organização proteica do ser humano. Tal como o corpo etérico se metamorfoseia de setênio em setênio, participando no desenvolvimento das capacidades anímico-espirituais de cada pessoa, a organização proteica também se transforma de setênio em setênio, por essa mesma razão, e, nessa transformação, participa intensamente o sistema imunológico. Alterações nas metamorfoses do corpo etérico se expressarão tanto para o lado orgânico, numa desregulação do sistema imunológico que pode determinar a aparição de doenças autoimunes, de evolução crônica, quanto nos quadros psicológicos característicos que podem apresentar esses mesmos pacientes, acompanhando o quadro da doença orgânica. O reconhecimento desses quadros em conjunto com o transtorno do desenvolvimento biográfico é de decisiva importância para o tratamento desses pacientes.(AU)

The human body is built and structured on its foundations by the etheric body. This occurs in part through the effect of the latter on the protein structure of the human body. The etheric body undergoes metamorphosis every seven years, modulating the development of the soul and spiritual capacities in each person. Similarly, the protein structure undergoes transformations through the same seven-year interval, with the immune system having an important role in this transformation. Metamorphosis in the etheric body may manifest both as an imbalance in the immune system (triggering chronic conditions such as auto-immune diseases) as well as specific psychological patterns of behavior that typically accompany such chronic diseases. The recognition of such manifestations as a facet of the personal biographical development is decisive in our ability to appropriately treat these patients.(AU)

Indomethacin induction of metamorphosis from the asexual stage to sexual stage in the moon jellyfish, Aurelia aurita.

We found while screening a chemical library that indomethacin, an inhibitor of prostaglandin biosynthesis, induced strobilation (metamorphosis from the asexual to sexual stage) in the moon jellyfish, Aurelia aurita. Indomethacin initiated strobilation in a dose-dependent manner, but was not involved in the progression of strobilation. Pharmacological experiments suggested that indomethacin could induce strobilation independently of prostaglandin biosynthesis.

Variety is the spice of life histories: comparison of intraspecific variability in marine invertebrates.

Life-history characteristics of marine invertebrates exhibit broad diversity across taxa as well as considerable variation within species. Although such variation is widely recognized, comparisons of the magnitude of variability as an outcome of evolutionary processes are relatively rare. Theory predicts, for example, that patterns of variability within and between clutches can arise as a consequence of population genetic structure, environmental variability, and the uncertainty of future ecological conditions. We review the strengths and weaknesses of several statistical methods for comparing variability across distributions, including Levene's test, use of the coefficient of variation in F-tests, and analysis of covariance. We then use four case studies from our own work and from the literature to illustrate adaptive patterns of variability related to metamorphosis, habitat differentiation, physiological stress, and life-history mode. These examples demonstrate the value of comparing variability for a range of questions associated with reproductive ecology, life-history biology, and genotype-by-environment interactions. We encourage researchers studying larval ecology and life-history evolution to explicitly consider the causes and consequences of variances in traits along with their means in models, experimental designs, analyses, and interpretations.

Effect of dietary probiotics on clownfish: a molecular approach to define how lactic acid bacteria modulate development in a marine fish.

We set out to determine whether probiotic addition would improve larval development in the false percula clownfish Amphiprion ocellaris and to determine what molecular responses could be observed in the larvae following probiotic exposure. We supplied the probiotic bacterial strain Lactobacillus rhamnosus IMC 501 to clownfish larvae from the first day posthatch simultaneously by live prey and with addition to rearing water (group 2) and exclusively by live prey (group 3). We observed twofold higher body weight in both clownfish larvae and juveniles when probiotics were supplied via live prey and added to the rearing water. In addition, development was accelerated with metamorphosis occurring 3 days earlier in fingerlings treated with probiotic. Alteration in molecular biomarkers supported the faster growth observation. There was significantly increased gene expression of factors involved in growth and development (insulin-like growth factors I and II, myostatin, peroxisome proliferator-activated receptors alpha and beta, vitamin D receptor alpha, and retinoic acid receptor gamma) when probiotics were delivered via live prey and added to the rearing water. Moreover, probiotic treatment lessened the severity of the general stress response as exhibited by lower levels of glucocorticoid receptor and 70-kDa heat shock protein gene expression. Furthermore, an improvement of skeletal head development was observed, with a 10-20% reduction in deformities for juveniles treated with probiotic. All data suggest a potent effect on development resulting from the administration of lactic acid bacteria to larval clownfish, and this study provides a preliminary molecular entry path into the investigation of mechanisms responsible for probiotic enhancement in fish development.

Embryonic development and metamorphosis of the scyphozoan Aurelia.

We investigated the development of Aurelia (Cnidaria, Scyphozoa) during embryogenesis and metamorphosis into a polyp, using antibody markers combined with confocal and transmission electron microscopy. Early embryos form actively proliferating coeloblastulae. Invagination is observed during gastrulation. In the planula, (1) the ectoderm is pseudostratified with densely packed nuclei arranged in a superficial and a deep stratum, (2) the aboral pole consists of elongated ectodermal cells with basally located nuclei forming an apical organ, which is previously only known from anthozoan planulae, (3) endodermal cells are large and highly vacuolated, and (4) FMRFamide-immunoreactive nerve cells are found exclusively in the ectoderm of the aboral region. During metamorphosis into a polyp, cells in the planula endoderm, but not in the ectoderm, become strongly caspase 3 immunoreactive, suggesting that the planula endoderm, in part or in its entirety, undergoes apoptosis during metamorphosis. The polyp endoderm seems to be derived from the planula ectoderm in Aurelia, implicating the occurrence of "secondary" gastrulation during early metamorphosis.

Treatment of highland frogs from the two-legged stage with homeopathically prepared thyroxin (10-11 - 10-21).

The influence of moderately diluted, agitated, i.e., homeopathically prepared, thyroxin solutions (10-11 - 10-21, final concentration in the basin water 0.6 10-15 - 0.6 10-25 parts by weight after the first application) on metamorphosis in highland Rana temporaria from the two-legged stage was studied. In accordance with the homeopathic idea of effects of specially prepared dilutions being inverse to those of their mother substances, animals were treated either with thyroxin 10-11 - 10-21 or analogously prepared blank solution (water). Development was monitored by documenting the number of animals that had entered the four-legged stage. It has been found that animals treated with the thyroxin solutions metamorphosed more slowly than the control animals, i.e., the effect of the homeopathically prepared thyroxin was opposed to the usual effect of molecular thyroxin. The number of test animals that reached the four-legged stage at defined points in time was smaller (2-13.5%) in the group treated with homeopathically prepared thyroxin at the points in time, compared to control. The results in this study sustain the previous multiresearcher findings that show that diluted homeopathically prepared thyroxin is able to slow down metamorphosis of R. temporaria.

The predominance of polyunsaturated fatty acids in the butterfly Morpho peleides before and after metamorphosis.

We hypothesized that the polyunsaturated fatty acids of the butterfly were probably derived from the diet and that there might be a great loss of body fat during metamorphosis. To substantiate these hypotheses, we analyzed the fatty acid composition and content of the diet, the larva, and the butterfly Morpho peleides. Both the diet and the tissues of the larva and butterfly had a high concentration of polyunsaturated fatty acids. In the diet, linolenic acid accounted for 19% and linoleic acid for 8% of total fatty acids. In the larva, almost 60% of the total fatty acids were polyunsaturated: linolenic acid predominated at 42% of total fatty acids, and linoleic acid was at 17%. In the butterfly, linolenic acid represented 36% and linoleic acid represented 11% of total fatty acids. The larva had a much higher total fatty acid content than the butterfly (20.2 vs. 6.9 mg). Our data indicate that the transformation from larva to butterfly during metamorphosis drastically decreased the total fatty acid content. There was bioenhancement of polyunsaturated fatty acids from the diet to the larva and butterfly. This polyunsaturation of membranes may have functional importance in providing membrane fluidity useful in flight.

Bioactivities of seven chrysanthemum species flowers powder on Spodoptera littoralis (Boiduval) larvae.

We have screened seven Chrysanthemum species, collected from different biotope in Tunisia for the antifeeding and growth regulatory activity against S. littoralis (Lepidoptera: Noctuidae) an insect pest of tomato. The flowers powder was added to the Poitout and Bues (1974) artificial diet at three concentrations: 4, 8, and 16%. After ten days of treatment, the consumed quantity determination revelled that these species have an antifeeding activity proportional to the concentration, more considerable for the C. fuscatum and C. Myconis flowers powder. At the concentration of 16% the larvae weight was significantly delayed in the average time to the control, these results were compared to the effect of Cestrum parquii on S. littoralis and Pieris brassicae The moulting date determination showed an elongation of the third, fourth and fifth stages at the high concentration probably due to the presence of juvenile hormones Analogues.

Control of head morphogenesis in an invertebrate asexually produced larva-like bud ( Cassiopea andromeda; Cnidaria: Scyphozoa).

Scyphopolyps of Cassiopea andromeda propagate asexually by forming larva-like buds which separate from the parent in a developmentally quiescent state. These buds metamorphose into sessile polyps when exposed to specific biogenic, chemical inducers. Morphogenesis of transversely dissected buds indicates the presence of pattern-determining signals; whereas the basal bud fragments may still form a complete scyphistoma the apical bud fragments develop spontaneously in the absence of an inducer into a polyp head without stalk and foot. Based on these findings Neumann (dissertation, Cologne University, 1980) postulated a head-inhibiting signal which is released at the basal pole and inhibits head formation at the apical end. Contrary to this hypothesis dissection itself might induce the development of head structures. The present study deals with the control of polyp head formation in C. andromeda. It concentrates on two points, namely the postulated head inhibitor and the involvement of compounds known to act during metamorphosis (the enzyme protein kinase C and the specific metamorphosis inducer Z-GPGGPA). We found that compared to intact buds and apical bud fragments transversely incised buds reached an intermediate stage of head development. This confirms Neumann's hypothesis. Consequently we focused on the mode of action and the chemical nature of the head-inhibiting signal in C. andromeda. Our results indicate that the head inhibitor may be included in one of six pooled fractions isolated from bud homogenate via gel filtration on a Sephadex G-50 column. The inhibitor is supposed to be water-soluble and to have a molecular weight of 850-1,500 Da. Furthermore we prove that head formation is not promoted by the metamorphosis-inducer Z-GPGGPA but is prevented by the inhibitors psychosine, chelerythrine and RO-32-0432 showing the involvement of protein kinase C in this process.

An endogenous peptide is involved in internal control of metamorphosis in the marine invertebrate Cassiopea xamachana (Cnidaria: Scyphozoa).

In the metagenetic life-cycle of the scyphozoan Cassiopea xamachana metamorphosis of planula-larvae or larva-like buds to polyps is triggered by specific external cues which are transmitted inside the larva or bud where internal signals finally coordinate the initiation of metamorphosis. This study deals with an endogenous metamorphosis inducer present in planulae and buds of Cassiopea. The inductive cue is localized in the basal part of the buds and can be characterized as a peptide with an apparent molecular weight of about 7,000 Da. Further purification was performed via reversed phase HPLC on a C18 column. Additional inhibitor assays revealed that protein kinase C and PI3 kinase, two known elements of the metamorphosis-inducing signal transduction cascade in Cassiopea, may act downstream of the endogenous inducing peptide.

Sterile polystyrene culture dishes induce transformation of polyps into medusae in Aurelia aurita (Scyphozoa, Cnidaria).

Polyps of Aurelia aurita produce medusae once a year. Under laboratory conditions this process, termed strobilation, can be induced by lowering the incubation temperature for about two weeks. Here we report a fast induction of strobilation by sterile polystyrene culture dishes. The effect is abolished when the culture dishes are washed twice with hot water prior to the experiment. We recommend that polystyrene cultureware should be pre-washed whenever there is an indication of unusual effects.

[The effect of food containing Cestrum paquii (Solanaceae) extract on various damaging Lepidoptera]. / L'impact de l'alimentation contenant du broyât de Cestrum paquii (Solanacées) sur des lépidoptères dommageables.

The repulsive activity of Cestrum parquii on the feeding behaviour of Pieris brassicae L. and Spodoptera littoralis Boisduval was studied. Antifeedant effect was observed at regular intervals during a week. Meal uptake was stopped after the first day, showing a strongly antifeedant effect of C. parquii extract on these two species. The impact of C. parquii with different concentrations of dry extract (2, 4, 8, 16, 32%) delayed the larval growth of Spodoptera littoralis Boisduval compared to untreated insects. Abnormal metamorphosis was observed with concentrations of 8 and 16% leading to death and abnormal morphogenesis. The antifeedant activity of C. parquii was also observed in S. littoralis and measured by the evolution of food uptake and the weight of larva. Effect seemed to be proportional to extract concentration in the diet. High concentration (32%) stopped growth in the earlier instar.