Gluconeogenesis in frogs during cooling and dehydration exposure: new insights into tissue plasticity of the gluconeogenic pathway dependent on abiotic factors.

Anurans undergo significant physiological changes when exposed to environmental stressors such as low temperatures and humidity. Energy metabolism and substrate management play a crucial role in their survival success. Therefore, understanding the role of the gluconeogenic pathway and demonstrating its existence in amphibians is essential. In this study, we exposed the subtropical frog Boana pulchella to cooling (-2.5°C for 24 h) and dehydration conditions (40% of body water loss), followed by recovery (24 h), and assessed gluconeogenesis activity from alanine, lactate, glycerol and glutamine in the liver, muscle and kidney. We report for the first time that gluconeogenesis activity by 14C-alanine and 14C-lactate conversion to glucose occurs in the muscle tissue of frogs, and this tissue activity is influenced by environmental conditions. Against the control group, liver gluconeogenesis from 14C-lactate and 14C-glycerol was lower during cooling and recovery (P<0.01), and gluconeogenesis from 14C-glutamine in the kidneys was also lower during cooling (P<0.05). In dehydration exposure, gluconeogenesis from 14C-lactate in the liver was lower during recovery, and that from 14C-alanine in the muscle was lower during dehydration (P<0.05). Moreover, we observed that gluconeogenesis activity and substrate preference respond differently to cold and dehydration. These findings highlight tissue-specific plasticity dependent on the nature of the encountered stressor, offering valuable insights for future studies exploring this plasticity, elucidating the importance of the gluconeogenic pathway and characterizing it in anuran physiology.

Urea and glucose modulation during freezing exposure in three temperate frogs reveals specific targets in relation to climate.

Amphibian diversity is most prominent in the warm and humid tropical and subtropical regions across the globe. Nonetheless, amphibians also inhabit high-altitude tropical mountains and regions at medium and high latitudes, exposing them to subzero temperatures and requiring behavioural or physiological adaptations to endure freezing events. While freeze tolerance has been predominantly reported in high-latitude zones where species endure prolonged freezing (several weeks or months), less is known about mid-latitudes amphibians exposed to occasional subzero temperatures. In this study, we employed a controlled ecological protocol, subjecting three frog species from the Iberian Peninsula (Rana parvipalmata, Epidalea calamita, and Pelobates cultripes) to a 2-h exposure to temperatures of -2 °C to investigate the accumulation of urea and glucose as physiological mechanisms associated with survival at freezing temperatures. Our results revealed a moderate response in the production of cryoprotectant metabolites under experimental freezing conditions, particularly urea, with notable findings in R. parvipalmata and E. calamita and no response in P. cultripes. However, no significant alterations in glucose concentrations were observed in any of the studied frog species. This relatively weak freezing tolerance response differs from the strong response exhibited by amphibians inhabiting high latitudes and enduring prolonged freezing conditions, suggesting potential reliance on behavioural adaptations to cope with occasional freezing episodes.

Metabolic changes in the subtropical frog Boana pulchella during experimental cooling and recovery conditions.

Frogs have developed biochemical and physiological adaptations to occupy diverse ecological niches on Earth successfully. Survival in frozen states is a fascinating strategy made possible by evolving adaptations to produce cryoprotectant solutes. The hylid frog Boana pulchella thrives in South American regions with cold climates, remaining active while enduring sporadic subzero temperatures during winter. The species' metabolic changes during subzero exposure remain unclear. Therefore, we exposed B. pulchella to cooling and recovery, assessing plasma and tissue metabolite changes. Cooling significantly reduced urea concentrations in plasma (P = 0.033), muscle (P = 0.001), heart (P = 0.009), and brain (P = 0.041) compared to acclimation. Liver glucose oxidation and glycogen synthesis were lower in cooling and recovery than in acclimation (P < 0.0001 and P = 0.0117, respectively). Muscle glycogen synthesis was lower in recovery than acclimation (P = 0.0249). These results demonstrate B. pulchella's physiological strategies during subzero exposure, likely reflecting species-specific evolutionary adaptations for brief subzero exposures that enable winter survival in its natural habitat.

The impact of chasing stress on the metabolism of the Atlantic Ghost Crab Ocypode quadrata (Fabricius, 1787).

Ocypode quadrata, a Ghost crab species found along the western Atlantic coast, is considered a bioindicator of anthropogenic impact on sandy beaches. Ghost Crabbing, a touristic activity in which ghost crabs are chased just for fun, is a potentially threatening activity for this crab. In crustaceans, metabolites such as glucose and lactate, and the gene expression of crustacean hyperglycemic hormone (CHH) and heat shock proteins (HSPs) increase when the animals are exposed to several types of stress, including alterations in temperature, salinity, or exposure to xenobiotics. This work was developed to identify if being chased by humans would affect these markers of stress in this species of crab. The effects of chasing stress on hemolymph and tissue metabolites and the gene expression levels of CHH and HSP70 were investigated. The levels of lactate in the hemolymph of stressed crabs were six times higher than those of control crabs immediately after chasing and decreased progressively during recovery, indicating an active anaerobic metabolism during the stress. On the contrary, glucose levels in the hemolymph of the stressed crabs increased progressively from 30 to 60 min after chasing, indicating an inverse correlation between glucose and lactate and the conversion of lactate to glucose by gluconeogenesis. In stressed crabs, the levels of triglycerides in the hemolymph decreased 30 min after chasing, while the opposite tended to occur in the hepatopancreas, indicating that during recovery, the crabs use triglycerides as energy source to sustain aerobic metabolism. Finally, this study demonstrates that ghost crabs are stressed by minimum human contact and that "ghost crabbing" must not be encouraged as a tourist activity.

Acute effects of a single moderate-intensity exercise bout performed in fast or fed states on cell metabolism and signaling: Comparison between lean and obese rats.

AIMS: Although the benefits of exercise can be potentiated by fasting in healthy subjects, few studies evaluated the effects of this intervention on the metabolism of obese subjects. This study investigated the immediate effects of a single moderate-intensity exercise bout performed in fast or fed states on the metabolism of gastrocnemius and soleus of lean and obese rats. MAIN METHODS: Male rats received a high-fat diet (HFD) for twelve weeks to induce obesity or were fed standard diet (SD). After this period, the animals were subdivided in groups: fed and rest (FER), fed and exercise (30 min treadmill, FEE), 8 h fasted and rest (FAR) and fasted and exercise (FAE). Muscle samples were used to investigate the oxidative capacity and gene expression of AMPK, PGC1α, SIRT1, HSF1 and HSP70. KEY FINDINGS: In relation to lean animals, obese animals' gastrocnemius glycogen decreased 60 %, triglycerides increased 31 %; glucose and alanine oxidation decreased 26 % and 38 %, respectively; in soleus, triglycerides reduced 46 % and glucose oxidation decreased 37 %. Exercise and fasting induced different effects in glycolytic and oxidative muscles of obese rats. In soleus, fasting exercise spared glycogen and increased palmitate oxidation, while in gastrocnemius, glucose oxidation increased. In obese animals' gastrocnemius, AMPK expression decreased 29 % and SIRT1 increased 28 % in relation to lean. The AMPK response was more sensitive to exercise and fasting in lean than obese rats. SIGNIFICANCE: Exercise and fasting induced different effects on the metabolism of glycolytic and oxidative muscles of obese rats that can promote health benefits in these animals.

Effects of tributyltin (TBT) on the intermediate metabolism of the crab Callinectes sapidus.

This study investigated if the exposure to tributyltin (TBT), a chemical used worldwide in boat antifouling paints, could result in metabolic disturbances in the blue crab Callinectes sapidus. After the exposure to TBT 100 or 1000 ng.L-1 for 48 and 96 h, hemolymph and tissues were collected to determine the concentration of metabolites and lipid peroxidation. The levels of glucose, lactate, cholesterol, and triglycerides in the hemolymph were not affected by TBT exposure. Hemolymph protein and heart glycogen increased in the crabs exposed to TBT 1000 for 96 h. Anterior gills protein and lipoperoxidation decreased after 96 h in all groups. These results suggest that C. sapidus can maintain energy homeostasis when challenged by the TBT exposure for 48 h and that metabolic alterations initiate after 96 h.

Anurans against SARS-CoV-2: A review of the potential antiviral action of anurans cutaneous peptides.

At the end of 2019, in China, clinical signs and symptoms of unknown etiology have been reported in several patients whose sample sequencing revealed pneumonia caused by the SARS-CoV-2 virus. COVID-19 is a disease triggered by this virus, and in 2020, the World Health Organization declared it a pandemic. Since then, efforts have been made to find effective therapeutic agents against this disease. Identifying novel natural antiviral drugs can be an alternative to treatment. For this reason, antimicrobial peptides secreted by anurans' skin have gained attention for showing a promissory antiviral effect. Hence, this review aimed to elucidate how and which peptides secreted by anurans' skin can be considered therapeutic agents to treat or prevent human viral infectious diseases. Through a literature review, we attempted to identify potential antiviral frogs' peptides to combat COVID-19. As a result, the Magainin-1 and -2 peptides, from the Magainin family, the Dermaseptin-S9, from the Dermaseptin family, and Caerin 1.6 and 1.10, from the Caerin family, are molecules that already showed antiviral effects against SARS-CoV-2 in silico. In addition to these peptides, this review suggests that future studies should use other families that already have antiviral action against other viruses, such as Brevinins, Maculatins, Esculentins, Temporins, and Urumins. To apply these peptides as therapeutic agents, experimental studies with peptides already tested in silico and new studies with other families not tested yet should be considered.

Seasonal variations in the intermediate metabolism in South American tree-frog Boana pulchella.

Seasonal metabolic changes can be observed in many anurans' species. In subtropical environments with environmental temperatures variations, the temperature is a factor that can influence the extent and intensity of activity in many anuran species. Nonetheless, some species of subtropical frogs may remain active throughout the year. Boana pulchella, a subtropical species, seems to be able to survive low temperatures and remain reproductively active even in the coldest months. Therefore, we hypothesized that B. pulchella presents seasonal changes in the energy metabolism to sustain activity during all year. This study evaluated the main energy substrate levels and metabolism of B. pulchella in plasma, liver and muscle of male individuals collected in winter, spring, summer and fall in the state of Rio Grande do Sul, Brazil. Our results showed that B. pulchella has a higher glycolytic oxidation rate in liver (P = 0.0152) and muscle (P = 0.0003) and higher glycogenesis from glucose in muscle (P = 0.0002) in summer, indicating the main energy substrates in this season is glucose. The higher muscle glycogen (P = 0.0008) and lower plasma glucose in fall (P = 0.0134) may indicate an anticipatory regulation for storing to the most thermally demanding cold period: winter. These results indicated seasonal differences in the main energy substrates, and these metabolic changes among seasons can be part of a metabolic adjustment allowing maintenance of reproductive activity all year in Boana pulchella species.