Effects of Desiccation on Metamorphic Climax in Bombina variegata: Changes in Levels and Patterns of Oxidative Stress Parameters

In this paper, we examined how the oxidative status (antioxidant system and oxidative damage) of Bombina variegata larvae changed during the metamorphic climax (Gosner stages: 42—beginning, 44—middle and 46—end) and compared the patterns and levels of oxidative stress parameters between individuals developing under constant water availability (control) and those developing under decreasing water availability (desiccation group). Our results revealed that larvae developing under decreasing water availability exhibited increased oxidative damage in the middle and end stages. This was followed by lower levels of glutathione in stages 44 and 46, as well as lower values of catalase, glutathione peroxidase, glutathione S-transferase and sulfhydryl groups in stage 46 (all in relation to control animals). Comparison between stages 42, 44 and 46 within treatments showed that individuals in the last stage demonstrated the highest intensities of lipid oxidative damage in both the control and desiccation groups. As for the parameters of the antioxidant system, control individuals displayed greater variety in response to changes induced by metamorphic climax than individuals exposed to desiccation treatment. The overall decrease in water availability during development led to increased oxidative stress and modifications in the pattern of AOS response to changes induced by metamorphic climax in larvae of B. variegata.

Remarkable metabolic reorganization and altered metabolic requirements in frog metamorphic climax

Background Metamorphic climax is the crucial stage of amphibian metamorphosis responsible for the morphological and functional changes necessary for transition to a terrestrial habitat. This developmental period is sensitive to environmental changes and pollution. Understanding its metabolic basis and requirements is significant for ecological and toxicological research. Rana omeimontis tadpoles are a useful model for investigating this stage as their liver is involved in both metabolic regulation and fat storage. Results We used a combined approach of transcriptomics and metabolomics to study the metabolic reorganization during natural and T3-driven metamorphic climax in the liver and tail of Rana omeimontis tadpoles. The metabolic flux from the apoptotic tail replaced hepatic fat storage as metabolic fuel, resulting in increased hepatic amino acid and fat levels. In the liver, amino acid catabolism (transamination and urea cycle) was upregulated along with energy metabolism (TCA cycle and oxidative phosphorylation), while the carbohydrate and lipid catabolism (glycolysis, pentose phosphate pathway (PPP), and β-oxidation) decreased. The hepatic glycogen phosphorylation and gluconeogenesis were upregulated, and the carbohydrate flux was used for synthesis of glycan units (e.g., UDP-glucuronate). In the tail, glycolysis, β-oxidation, and transamination were all downregulated, accompanied by synchronous downregulation of energy production and consumption. Glycogenolysis was maintained in the tail, and the carbohydrate flux likely flowed into both PPP and the synthesis of glycan units (e.g., UDP-glucuronate and UDP-glucosamine). Fatty acid elongation and desaturation, as well as the synthesis of bioactive lipid (e.g., prostaglandins) were encouraged in the tail during metamorphic climax. Protein synthesis was downregulated in both the liver and tail. The significance of these metabolic adjustments and their potential regulation mechanism are discussed. Conclusion The energic strategy and anabolic requirements during metamorphic climax were revealed at the molecular level. Amino acid made an increased contribution to energy metabolism during metamorphic climax. Carbohydrate anabolism was essential for the body construction of the froglets. The tail was critical in anabolism including synthesizing bioactive metabolites. These findings increase our understanding of amphibian metamorphosis and provide background information for ecological, evolutionary, conservation, and developmental studies of amphibians.

Remarkable metabolic reorganization and altered metabolic requirements in frog metamorphic climax

Background Metamorphic climax is the crucial stage of amphibian metamorphosis responsible for the morphological and functional changes necessary for transition to a terrestrial habitat. This developmental period is sensitive to environmental changes and pollution. Understanding its metabolic basis and requirements is significant for ecological and toxicological research. Rana omeimontis tadpoles are a useful model for investigating this stage as their liver is involved in both metabolic regulation and fat storage. Results We used a combined approach of transcriptomics and metabolomics to study the metabolic reorganization during natural and T3-driven metamorphic climax in the liver and tail of Rana omeimontis tadpoles. The metabolic flux from the apoptotic tail replaced hepatic fat storage as metabolic fuel, resulting in increased hepatic amino acid and fat levels. In the liver, amino acid catabolism (transamination and urea cycle) was upregulated along with energy metabolism (TCA cycle and oxidative phosphorylation), while the carbohydrate and lipid catabolism (glycolysis, pentose phosphate pathway (PPP), and β-oxidation) decreased. The hepatic glycogen phosphorylation and gluconeogenesis were upregulated, and the carbohydrate flux was used for synthesis of glycan units (e.g., UDP-glucuronate). In the tail, glycolysis, β-oxidation, and transamination were all downregulated, accompanied by synchronous downregulation of energy production and consumption. Glycogenolysis was maintained in the tail, and the carbohydrate flux likely flowed into both PPP and the synthesis of glycan units (e.g., UDP-glucuronate and UDP-glucosamine). Fatty acid elongation and desaturation, as well as the synthesis of bioactive lipid (e.g., prostaglandins) were encouraged in the tail during metamorphic climax. Protein synthesis was downregulated in both the liver and tail. The significance of these metabolic adjustments and their potential regulation mechanism are discussed. Conclusion The energic strategy and anabolic requirements during metamorphic climax were revealed at the molecular level. Amino acid made an increased contribution to energy metabolism during metamorphic climax. Carbohydrate anabolism was essential for the body construction of the froglets. The tail was critical in anabolism including synthesizing bioactive metabolites. These findings increase our understanding of amphibian metamorphosis and provide background information for ecological, evolutionary, conservation, and developmental studies of amphibians.

Remarkable metabolic reorganization and altered metabolic requirements in frog metamorphic climax

BackgroundMetamorphic climax is the crucial stage of amphibian metamorphosis responsible for the morphological and functional changes for landing. This developmental period is most sensitive to environmental changes and pollution. Understanding its metabolic basis and requirements has great significance to ecological and toxicological researches. Rana omeimontis tadpoles are unique model for investigating this issue, as their liver has the dual role of metabolic regulation and fat storage.ResultsUsing a combined approach of transcriptomics and metabolomics, we revealed the metabolic reorganization during natural and T3-driven metamorphic climax by reconstructing the metabolic networks in the liver and tail of Rana omeimontis tadpoles. In metamorphic tadpoles, the metabolic flux from the apoptotic tail replaced hepatic fat storage as metabolic fuel, resulting in increased hepatic amino acid and fat level. In the liver, amino acid catabolism (transamination and urea cycle) was upregulated along with energy metabolism (TCA cycle and oxidative phosphorylation), while the carbohydrate and lipid catabolism (glycolysis, pentose phosphate pathway/PPP and β-oxidation) were decreased. Meanwhile, the hepatic glycogen phosphorylation and gluconeogenesis were upregulated, and the carbohydrate flux likely flowed into the synthesis of glycan units (e.g., UDP-glucuronate). In the tail, glycolysis, β-oxidation, and transamination were all downregulated, accompanied by synchronous downregulation of energy production and consumption. The glycogenolysis was maintained in the tail, and the carbohydrate flux likely flowed into both PPP and the synthesis of glycan units (e.g., UDP-glucuronate and UDP-glucosamine). Interestingly, the fatty acid elongation and desaturation, as well as the synthesis of bioactive lipid (e.g., prostaglandins) were encouraged in the tail during metamorphic climax. Protein synthesis was downregulated in both the liver and tail. The significance of these metabolic adjustments and their potential regulation mechanism were discussed.ConclusionThe energic strategy and anabolic requirements during metamorphic climax were revealed at molecular level. Amino acid has increased contribution to energy metabolism during metamorphic climax. Carbohydrate anabolism is more intensively required than protein synthesis for the body construction of the froglet. The tail plays a critical role in anabolism and even metabolic regulation. These findings would deepen our understanding on the requirement and proceeding of amphibian metamorphosis.

Endocrine Disruption Alters Developmental Energy Allocation and Performance in Rana temporaria

Environmental change exposes wildlife to a wide array of environmental stressors that arise from both anthropogenic and natural sources. Many environmental stressors with the ability to alter endocrine function are known as endocrine disruptors, which may impair the hypothalamus–pituitary–thyroid axis resulting in physiological consequences to wildlife. In this study, we investigated how the alteration of thyroid hormone (TH) levels due to exposure to the environmentally relevant endocrine disruptor sodium perchlorate (SP; inhibitory) and exogenous L-thyroxin (T4; stimulatory) affects metabolic costs and energy allocation during and after metamorphosis in a common amphibian (Rana temporaria). We further tested for possible carry-over effects of endocrine disruption during larval stage on juvenile performance. Energy allocated to development was negatively related to metabolic rate and thus, tadpoles exposed to T4 could allocate 24% less energy to development during metamorphic climax than control animals. Therefore, the energy available for metamorphosis was reduced in tadpoles with increased TH level by exposure to T4. We suggest that differences in metabolic rate caused by altered TH levels during metamorphic climax and energy allocation to maintenance costs might have contributed to a reduced energetic efficiency in tadpoles with high TH levels. Differences in size and energetics persisted beyond the metamorphic boundary and impacted on juvenile performance. Performance differences are mainly related to strong size-effects, as altered TH levels by exposure to T4 and SP significantly affected growth and developmental rate. Nevertheless, we assume that juvenile performance is influenced by a size-independent effect of achieved TH. Energetic efficiency varied between treatments due to differences in size allocation of internal macronutrient stores. Altered TH levels as caused by several environmental stressors lead to persisting effects on metamorphic traits and energetics and, thus, caused carry-over effects on performance of froglets. We demonstrate the mechanisms through which alterations in abiotic and biotic environmental factors can alter phenotypes at metamorphosis and reduce lifetime fitness in these and likely other amphibians.

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.

Is there a time lag between the metamorphism and emplacement of plutons in the Axial Zone of the Pyrenees?

Detailed petrographic and geochemical studies conducted on zircons from the Lys-Caillaouas pluton reveal their igneous and metamorphic affinities. The igneous zircons constrain the emplacement of the pluton to 300±2 Ma. By contrast, the metamorphic zircons yield an older age of 307±3 Ma, which probably dates the thermal peak of the HT/LP Variscan metamorphism. Therefore, a short time lag of c. 7 Ma emerges between the metamorphic climax and emplacement of the pluton in the Axial Zone (Pyrenees).

Comparative sensitivities of larval stages of the cane toad, Rhinella marina, and the striped marsh frog, Limnodynastes peronii, to atrazine

Variations in larval sensitivities to atrazine were determined in the Australian native striped marsh frog, Limnodynastes peronii, and the introduced cane toad, Rhinella marina. The static acute test design involved six nominal concentrations of atrazine, including control, solvent control, 3, 6, 12, and 24 mg L–1. Gosner stages 22–23 as hatchlings, stages 25–26, 28–29, and 32–33 as premetamorphic, 36–37 as prometamorphic and 40–41 as metamorphic climax stages of cane toads and the first four sets of Gosner stages of striped marsh frogs were exposed to atrazine treatments for 96 h. Results showed that late larval stages were more sensitive than early stages and different premetamorphic stages showed variations in sensitivities in both test species. The striped marsh frog showed a stronger concentration- and stage-dependent response and greater sensitivity to atrazine than the cane toad. In both experimental species, Gosner stages 28–29 showed better concentration-dependent increase in sensitivities to atrazine compared with other larval stages. It can be concluded that inter- and intra-species variations in sensitivities to atrazine may occur in Australian anurans and native species may show greater sensitivity to acute concentrations of atrazine than the introduced cane toad.

Anuran life history plasticity: Variable practice in determining the end-point of larval development

Plasticity in the timing of life history events and their impact on individual fitness, particularly the timing of and size at metamorphosis in animals with complex life cycles such as anuran amphibians, has long been of interest to ecologists. For different studies on life history plasticity to be comparable, there must be clearly defined and commonly agreed transition points, but it is unclear how consistently this is being performed in studies using anuran amphibians. In a review of 157 published studies, I found considerable variation in defining the end point of the larval phase. While a slight majority used the emergence of the forelimbs as the conclusion of the larval phase, some used a period within the developmental phase of metamorphic climax and others used the resorption of the tail. Studies included in this review, that assessed the same life history variable at two different developmental stages, reported some differences in results depending on which developmental stage was used. Recent evidence also shows that metamorphic climax is itself a period which can vary with environmental conditions, but, even in studies that included part or all of metamorphic climax in the larval phase, the treatment of individuals during metamorphic climax was not reported. Therefore, I argue that life history studies on anuran amphibians should distinguish the following phases: larval, metamorphic climax, juvenile, adult; that the end of the larval phase is best defined in ecological studies by forelimb emergence and that conditions under which individuals undergo metamorphic climax should be fully described.