Frog Tissues Research Articles

Isolation and Characterization of a Frog Virus 3 Strain from a Wood Frog (Rana sylvatica) in Wood Buffalo National Park.

Members of the Iridoviridae family, genus Ranavirus, represent a group of globally emerging pathogens of ecological and economic importance. In 2017, an amphibian die-off of wood frogs (Rana sylvatica) and boreal chorus frogs (Pseudacris maculata) was reported in Wood Buffalo National Park, Canada. Isolation and complete genomic sequencing of the tissues of a wood frog revealed the presence of a frog virus 3 (FV3)-like isolate, Rana sylvatica ranavirus (RSR), with a genome size of 105,895 base pairs, 97 predicted open reading frames (ORFs) bearing sequence similarity to FV3 (99.98%) and a FV3-like isolate from a spotted salamander in Maine (SSME; 99.64%). Despite high sequence similarity, RSR had a unique genomic composition containing ORFs specific to either FV3 or SSME. In addition, RSR had a unique 13 amino acid insertion in ORF 49/50L. No differences were found in the in vitro growth kinetics of FV3, SSME, and RSR; however, genomic differences between these isolates were in non-core genes, implicated in nucleic acid metabolism and immune evasion. This study highlights the importance of viral isolation and complete genomic analysis as these not only provide information on ranavirus spatial distribution but may elucidate genomic factors contributing to host tropism and pathogenicity.

Small RNA and Freeze Survival: The Cryoprotective Functions of MicroRNA in the Frozen Muscle Tissue of the Grey Tree Frog.

The grey tree frog, Dryophytes versicolor, survives whole-body freezing for weeks during cold winter months. Survival in a state devoid of available food, water, or oxygen forces a reliance on metabolic rate depression (MRD) and the reprioritization of bodily functions. This study utilizes next-generation sequencing (NGS) and bioinformatic analyses to characterize changes in the microRNAome of D. versicolor. When comparing control to frozen groups, five microRNAs (miRNA) were found to be differentially regulated (miR-143-3p, miR-30e-3p, miR-10a-5p, miR-140-3p, and miR-148a-3p), suggesting that they play key roles in freeze survival. The KEGG and GO analyses of these changes predicted a significant negative enrichment of terms associated with cell proliferation and active metabolism while simultaneously predicting the upregulation of cell signalling terms. These results suggest a fast-acting regulatory role for miRNA in contributing to the reorganization of gene expression and the limitation of energy-expensive processes during MRD in the hind leg skeletal muscle of the frog.

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.

Tissue-Specific Distribution and Maternal Transfer of Persistent Organic Halogenated Pollutants in Frogs.

Persistent organic pollutants pose a great threat to amphibian populations, but information on the bioaccumulation of contaminants in amphibians remains scarce. To examine the tissue distribution and maternal transfer of organic halogenated pollutants (OHPs) in frogs, seven types of tissues from black-spotted frog (muscle, liver, kidney, stomach, intestine, heart, and egg) were collected from an e-waste-polluted area in South China. Among the seven frog tissues, median total OHP concentrations of 2.3 to 9.7 μg/g lipid weight were found (in 31 polychlorinated biphenyl [PCB] individuals and 15 polybrominated diphenyl ether [PBDE], dechlorane plus [syn-DP and anti-DP], bexabromobenzene [HBB], polybrominated biphenyl] PBB153 and -209], and decabromodiphenyl ethane [DBDPE] individuals). Sex-specific differences in contaminant concentration and compound compositions were observed among the frog tissues, and eggs had a significantly higher contaminant burden on the whole body of female frogs. In addition, a significant sex difference in the concentration ratios of other tissues to the liver was observed in most tissues except for muscle. These results suggest that egg production may involve the mobilization of other maternal tissues besides muscle, which resulted in the sex-specific distribution. Different parental tissues had similar maternal transfer mechanisms; factors other than lipophilicity (e.g., molecular size and proteinophilic characteristics) could influence the maternal transfer of OHPs in frogs. Environ Toxicol Chem 2024;43:1557-1568. © 2024 SETAC.

Larger Common River Frogs (Amietia delalandii) have Fewer and Shorter Tissue Microplastic Fibres than Smaller Frogs

Little is known about microplastics (MPs) in adult frogs. We investigated MPs in adult Common River Frogs (Amietia delalandii) from Potchefstroom, South Africa. Five kinds of samples were analysed: natural water, water used to rinse the skin, skin, intestine, and the remainder of the body (corpus). Tissues were digested. Microplastics occurred in all frogs and sample types (1128 MPs counted). Fibres were the most prevalent MP. Fibre lengths were between 28 and 4300 μm, either polyester or polyvinyl alcohol. MPs in skin were likely derived from the ambient, and MPs in the corpus from translocation via the skin. Fibres in tissues were significantly shorter in larger frogs, a phenomenon we provisionally assign to in situ biodegradation. Microplastics in frogs can potentially be transferred through the food web to higher trophic levels. This study provides the first evidence of MPs in adult frog tissues and avenues for further investigations.Graphical

Investigation of heavy metals in tissues and habitats of three edible frogs from Türkiye.

The soil, water, and organisms have been contaminated by heavy metals due to human activities and industrialization, which has produced a major environmental problem that has a deleterious effect on human health and food quality. Frogs, one of the good bioindicators for environmental pollution, are also among the alternative essential protein sources for humans. In Türkiye, three of these frogs are edible: Pelophylax ridibundus, Pelophylax bedriagae, and Pelophylax caralitanus, also known as Anatolian water frogs. Hence, to assess the possible health risks that might result from consuming frog legs in addition aquatic habitat of Anatolian water frogs, the water, sediment, and frog tissue samples (muscle and liver) were obtained from 11 different provinces covering all regions of Türkiye and analyzed to determine Cd, Cu, Cr, Zn, Pb, and As concentrations. The results revealed considerable variations in heavy metal concentrations among frog tissues, influenced by the sampling sites and species (ANOVA: p < 0.05). The Estimated Daily Intake (EDI) values, calculated based on the average serving size, were also lower than the Provisional Tolerable Daily Intake (PTDI) levels for adult consumers. Furthermore, the study computed the Target Hazard Quotient (THQ) values for heavy metals, all of which were below the critical value of 1, indicating that consuming the hind leg muscles from these frog species would not pose an adverse health risk for humans.

THE CATALYTIC PROPERTIES OF LACTATE DEHYDROGENASE IN SKELETAL MUSCLES AND LIVER OF THE MARSH FROG (PELOPHYLAX RIDIBUNDUS) DEPEND ON ITS ECOLOGICAL AND GEOGRAPHICAL LOCATION

The marsh frog (Pelophylax ridibundus) has a widespread distribution range, which is due to a variety of adaptations that contribute to the development of tolerance to a wide range of physicochemical environmental factors. Of particular interest are the adaptations of these animals to different levels of oxygen in mid- and high-altitude conditions. In this work, a comparative analysis of the kinetic parameters of lactate dehydrogenase (LDH) in the liver of marsh frogs living in the mountainous and lowland regions of Dagestan was carried out. Animals caught in their habitats were decapitated, the liver and calf muscles were isolated, and they were placed in liquid nitrogen. In the laboratory, the selected tissues were homogenized and mitochondria-free cytosol was obtained by differential centrifugation, in which LDH activity was determined. It was found that LDH activity is significantly higher in the tissues of frogs from mountainous regions: by 42.4% in the muscles and 2.38 times in the liver (p 0.05). The high efficiency of catalysis is ensured due to significant changes in the catalytic parameters of the enzyme: an increase in Vmax (50.9% in muscles and 70% in the liver (p 0.05)) and a decrease in Km. (45.9% in muscles and 69% in liver, (p 0.05)). A more pronounced difference, compared to muscles, between LDH activity in the liver of foothill and lowland populations of frogs suggests that the sensitivity of liver LDH to changes in oxygen tension is higher. The vector of a number of other kinetic parameters of LDH (Ki, Sopt, Δ) in the liver of animals from mountainous landscapes is absolutely opposite to that of skeletal muscles. High activity and modifications of the catalytic properties of LDH in the tissues of marsh frogs living in mid-mountain areas may play an important role in the adaptation of these animals to conditions of oxygen deficiency.

Effect of khat leaves extract (Catha edulis) on the excitability of frog gastrocnemius.

This study was held on the isolated excitable tissue of frog's gastrocnemius muscle. Experiments were down by determining the dose dependent of leaves of khat extract. Work down in accordance with personal presupposition since reported that khat have psycho stimulant effects accordingly we thought khat might have effects on systemically biology that mediated by chticol amines. Results showed the effect of khat leaves extract on excitability of muscle contraction that was easily attested when the electrical stimuli were applied either through the nerve or directly to the muscle. Whereas khat treatment showed a significant increase in the contraction period of the gastrocnemius muscle and both of the relaxation periods and half relaxation time were shortened. Also it was found that pretreatment by khat leaves extract caused changes in the tension output, increased the height of effect intensity with prolonged the contraction period, while the latent period was decreased and reduced slightly the half relaxation period of the twitch comparison with the normal control muscle. These experimental results indicated that khat had highly stimulant effects on muscle, whereas caused liberation of thermoenzymatic and chemical energy during contraction which converted into mechanical energy, which manifested its self as work.

Hypoxia inducible factor-1α responds to freezing, anoxia and dehydration stresses in a freeze-tolerant frog

The wood frog, Rana sylvatica (aka Lithobates sylvaticus) is the main model for studies of natural freeze tolerance among amphibians living in seasonally cold climates. During freezing, ∼65% of total body water can be converted to extracellular ice and this imposes both dehydration and hypoxia/anoxia stresses on cells. The current study analyzed the responses of the alpha subunit of the hypoxia-inducible transcription factor (HIF-1), a crucial oxygen-sensitive regulator of gene expression, to freezing, anoxia or dehydration stresses, examining six tissues of wood frogs (liver, skeletal muscle, brain, heart, kidney, skin). RT-PCR revealed a rapid elevation hif-1α transcript levels within 2 h of freeze initiation in both liver and brain and elevated levels of both mRNA and protein in liver and muscle after 24 h frozen. However, both transcript and protein levels reverted to control values after thawing except for HIF-1 protein in liver that dropped to ∼60% of control. Independent exposures of wood frogs to anoxia or dehydration stresses (two components of freezing) also triggered upregulation of hif-1α transcripts and/or HIF-1α protein in liver and kidney with variable responses in other tissues. The results show active modulation of HIF-1 in response to freezing, anoxia and dehydration stresses and implicate this transcription factor as a contributor to the regulation of metabolic adaptations needed for long term survival of wood frogs in the ischemic frozen state.

Ova kurbağası, Pelophylax ridibundus'un (Pallas, 1771) bacak tüketiminden elde edilen eser element biyobirikimi ve sağlık riski değerlendirmesi

Amphibians, which can live in aquatic and terrestrial environments, are a good indicator of pollution in these areas. Although frog leg is not consumed frequently in some cuisines, including Turkey, it is important in terms of evaluating metal accumulation since it is preferred as human food in many European countries. In this study, the quantities of trace elements (Cd, Pb, Cu, Zn, As, Co, Cr, Ni, Mn, V) were measured in the edible tissues (muscles) of an amphibian species by sampling from two frog farms in Turkey. It was aimed to assess possible health hazards for humans by frog legs consumption comparing with the toxicological limit values, including provisional tolerable weekly intake (PTWI), target hazard quotient (THQ), and Hazard Index (HI). In general, the average values (µg kg−1) of trace elements were Zn (3.437.62)&gt; Pb (69.22)&gt; Cu (66.72)&gt; Mn (35.07)&gt; As (24.24)&gt; Cr (11.47)&gt; Ni (6.94)&gt; Cd (6.51)&gt; Co (2.97)&gt; V (&lt;0.001). The results indicated that concentrations of the analyzed trace elements were determined below the European Commission’s permitted levels and edible tissues of the marsh frog posed no carcinogenic health risk to humans.

Per- and Polyfluoroalkyl Substances (PFASs) Impair Lipid Metabolism in Rana nigromaculata: A Field Investigation and Laboratory Study.

Per- and polyfluoroalkyl substances (PFASs) are ubiquitous environmental pollutants, causing environmental threats and public health concerns, but information regarding PFAS hepatotoxicity remains elusive. We investigated the effects of PFASs on lipid metabolism in black-spotted frogs through a combined field and laboratory study. In a fluorochemical industrial area, PFASs seriously accumulate in frog tissues. PFAS levels in frog liver tissues are positively related to the hepatosomatic index along with triglyceride (TG) and cholesterol (TC) contents. In the laboratory, frogs were exposed to 1 and 10 μg/L PFASs, respectively (including PFOA, PFOS, and 6:2 Cl-PFESA). At 10 μg/L, PFASs change the hepatic fatty acid composition and significantly increase the hepatic TG content by 1.33 to 1.87 times. PFASs induce cross-talk accumulation of TG, TC, and their metabolites between the liver and serum. PFASs can bind to LXRα and PPARα proteins, further upregulate downstream lipogenesis-related gene expression, and downregulate lipolysis-related gene expression. Furthermore, lipid accumulation induced by PFASs is alleviated by PPARα and LXRα antagonists, suggesting the vital role of PPARα and LXRα in PFAS-induced lipid metabolism disorders. This work first reveals the disruption of PFASs on hepatic lipid homeostasis and provides novel insights into the occurrence and environmental risk of PFASs in amphibians.

Regulation of the unfolded protein response during dehydration stress in African clawed frogs, Xenopus laevis.

The unfolded protein response (UPR) is a wide-ranging cellular response to accumulation of malfolded proteins in the endoplasmic reticulum (ER) and acts as a quality control mechanism to halt protein processing and repair/destroy malfolded proteins under stress conditions of many kinds. Among vertebrate species, amphibians experience the greatest challenges in maintaining water and osmotic balance, the high permeability of their skin making them very susceptible to dehydration and challenging their ability to maintain cellular homeostasis. The present study evaluates the involvement of the UPR in dealing with dehydration-mediated disruption of protein processing in the tissues of African clawed frogs, Xenopus laevis. This primarily aquatic frog must deal with seasonal drought conditions in its native southern Africa environment. Key markers of cellular stress that impact protein processing were identified in six tissues of frogs that had lost 28% of total body water, as compared with fully hydrated controls. This included upregulation of glucose-regulated proteins (GRPs) that are resident chaperones in the ER, particularly 2-ninefold increases in GRP58, GRP75, and/or GRP94 in the lung and skin. Activating transcription factors (ATF3, ATF4, ATF6) that mediate UPR responses also responded to dehydration stress, particularly in skeletal muscle where both ATF3 and ATF4 rose strongly in the nucleus. Other protein markers of the UPR including GADD34, GADD153, EDEM, and XBP-1 also showed selective upregulation in frog tissues in response to dehydration and nuclear levels of the transcription factors XBP-1 and P-CREB rose indicating up-regulation of genes under their control.

Histopathology of Endocrine Organs of Miranda's White-Lipped Frogs (Leptodactylus macrosternum) from Cultivated and Non-Cultivated Regions in Semi-Arid Northeastern Brazil

Endocrine disruption defines the influence of environmental contaminants on the control of behaviour, growth and reproduction of free-living species. Amphibians are considered as bioindicators of environmental balance in aquatic and terrestrial systems and of physiological and immunological sensitivity to environmental stressors. In the Caatinga Domain of northeastern Brazil, the Lower Jaguaribe River (LJR) region is outstanding for the conservation of anurans and has considerable species diversity. However, the use of pesticides and chemical fertilisers is of increasing concern because of consequent risks to environmental, human and animal health. Miranda's white-lipped frogs (Leptodactylus macrosternum) are abundant and well-adapted in this region and, therefore, are considered to be a suitable bioindicator. The objective of this work was to evaluate the morphology of endocrine organs of this species for evidence of potential impacts of agrosystems. We found significantly smaller females and parathyroid cytolysis in frogs collected from cultivated areas, compared with those from non-cultivated regions. Interrenal degeneration was found in both sample populations. Chemical analysis detected pesticides and potentially toxic elements in tissues of Miranda's white-lipped frog tissues from the LJR region (data not shown). These findings suggest that the parathyroid gland acts as a biomarker of environmental toxicity in cultivated areas in the LJR region. In this study, the other examined endocrine organs did not show significant potential for use in biomonitoring the impact of toxic chemicals.

Bioaccumulation of Trace Elements and Health Risk Predictions in Edible Tissues of the Marsh Frog.

Certain amphibian species have long served as a valuable protein source for humans, in addition to being good bioindicators for environmental pollutants. Hence, to investigate the consumption outcomes leading to potential health risks, we determined the trace element (TE) levels in the hind leg and liver tissues of marsh frogs (Pelophylax ridibundus), one of the delicacies of several cuisines today. The sediment, water, and frog tissue samples were collected from 15 different locations of NE Turkey and analyzed to determine the arsenic (As), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), manganese (Mn), nickel (Ni), lead (Pb), vanadium (V), and zinc (Zn) concentrations. The TE concentrations in the sediment, water, and individuals were detected to show significant variations among sampling stations (p < 0.05). Yet, Cd and Pb concentrations of the hind legs cooked and enjoyed in the diets were determined below the European Commission's permitted levels. Furthermore, based on the TEs in edible tissues, consumption of the marsh frog did not appear to pose a risk to humans in terms of provisional tolerable weekly intake (PTWI), target hazard coefficient (THQ), and hazard index (HI).

Concentration of current-use pesticides in frogs from the Pampa region and correlation of a mixture toxicity index with biological effects

Contamination with current-use pesticides is frequently mentioned as a key factor in global amphibian declines although a limited number of studies have examined the mixture of pesticides accumulated by free-living frogs. This study examined the presence of 46 different pesticide residues in the muscle and kidney tissues of two frog species living in close association with row crops in the Pampa region of Argentina: The terrestrial Leptodactylus latinasus and the semi-aquatic Leptodactylus latrans. A total of 20 different pesticides were identified in frog tissues; chlorpyrifos-methyl, pirimiphos-methyl and acetochlor being the most frequently detected molecules. Overall, one or more pesticide residues (up to 12 in a single frog) were detected in 40–57 % of L. latrans. L. latinasus was found to present more pesticide detections than L. latrans. Interestingly, frog sampled in a natural reserve where no pesticides are applied presented an equivalent frequency of detections as frogs living near a crop. In L. latrans, the calculation of a pesticide toxicity index (PTI) permitted to highlight the existence of a strong positive correlation between PTI and liver GSH contents of females whereas, in males, PTI was negatively correlated with the perimeter of testicular seminiferous tubules. Males also presented near significant negative correlations between PTI and both body condition and the scaled fat index. These results indicate that frogs inhabiting agricultural regions are exposed to a complex and diffuse contamination by pesticide mixtures which is likely responsible for a number of biological effects that may be relevant at the population level.

Sub-lethal exposure to malaria vector control pesticides causes alterations in liver metabolomics and behaviour of the African clawed frog (Xenopus laevis)

In this study we explore the sub-lethal effects of two malaria vector control pesticides, deltamethrin and dichlorodiphenyltrichloroethane (DDT), on Xenopus laevis by incorporating different levels of biological organisation. Pesticide accumulation in frog tissue was measured alongside liver metabolomics and individual swimming behaviour to assess whether changes presented at these different levels, and if such changes could be linked between levels. Results showed evidence of concentration dependent accumulation of DDT and its metabolites, but no measurable accumulation of deltamethrin in adult X. laevis after 96 h of exposure. Both DDT and deltamethrin were shown to cause alterations in the liver metabolome of X. laevis. We also showed that some of these changes can be enhanced in exposure to a mixture of these two pesticides. Initial behavioural responses recorded directly after exposure were seen in the form of decreased activity, less alterations between mobility states, and less time spent at the water surface. This response persisted after 96 h of exposure to a mixture of the two pesticides. This study shows that sub-lethal exposure to pesticides can alter the biochemical homeostasis of frogs with the potential to cascade onto behavioural and ecological levels in mixture exposure scenarios.

Virus-Targeted Transcriptomic Analyses Implicate Ranaviral Interaction with Host Interferon Response in Frog Virus 3-Infected Frog Tissues.

Ranaviruses (Iridoviridae), including Frog Virus 3 (FV3), are large dsDNA viruses that cause devastating infections globally in amphibians, fish, and reptiles, and contribute to catastrophic amphibian declines. FV3’s large genome (~105 kb) contains at least 98 putative open reading frames (ORFs) as annotated in its reference genome. Previous studies have classified these coding genes into temporal classes as immediate early, delayed early, and late viral transcripts based on their sequential expression during FV3 infection. To establish a high-throughput characterization of ranaviral gene expression at the genome scale, we performed a whole transcriptomic analysis (RNA-Seq) using total RNA samples containing both viral and cellular transcripts from FV3-infected Xenopus laevis adult tissues using two FV3 strains, a wild type (FV3-WT) and an ORF64R-deleted recombinant (FV3-∆64R). In samples from the infected intestine, liver, spleen, lung, and especially kidney, an FV3-targeted transcriptomic analysis mapped reads spanning the full-genome coverage at ~10× depth on both positive and negative strands. By contrast, reads were only mapped to partial genomic regions in samples from the infected thymus, skin, and muscle. Extensive analyses validated the expression of almost all of the 98 annotated ORFs and profiled their differential expression in a tissue-, virus-, and temporal class-dependent manner. Further studies identified several putative ORFs that encode hypothetical proteins containing viral mimicking conserved domains found in host interferon (IFN) regulatory factors (IRFs) and IFN receptors. This study provides the first comprehensive genome-wide viral transcriptome profiling during infection and across multiple amphibian host tissues that will serve as an instrumental reference. Our findings imply that Ranaviruses like FV3 have acquired previously unknown molecular mimics, interfering with host IFN signaling during evolution.

Targeted Transcriptomics of Frog Virus 3 in Infected Frog Tissues Reveal Non-Coding Regulatory Elements and microRNAs in the Ranaviral Genome and Their Potential Interaction with Host Immune Response.

BackgroundFrog Virus 3 (FV3) is a large dsDNA virus belonging to Ranaviruses of family Iridoviridae. Ranaviruses infect cold-blood vertebrates including amphibians, fish and reptiles, and contribute to catastrophic amphibian declines. FV3 has a genome at ~105 kb that contains nearly 100 coding genes and 50 intergenic regions as annotated in its reference genome. Previous studies have mainly focused on coding genes and rarely addressed potential non-coding regulatory role of intergenic regions.ResultsUsing a whole transcriptomic analysis of total RNA samples containing both the viral and cellular transcripts from FV3-infected frog tissues, we detected virus-specific reads mapping in non-coding intergenic regions, in addition to reads from coding genes. Further analyses identified multiple cis-regulatory elements (CREs) in intergenic regions neighboring highly transcribed coding genes. These CREs include not only a virus TATA-Box present in FV3 core promoters as in eukaryotic genes, but also viral mimics of CREs interacting with several transcription factors including CEBPs, CREBs, IRFs, NF-κB, and STATs, which are critical for regulation of cellular immunity and cytokine responses. Our study suggests that intergenic regions immediately upstream of highly expressed FV3 genes have evolved to bind IRFs, NF-κB, and STATs more efficiently. Moreover, we found an enrichment of putative microRNA (miRNA) sequences in more than five intergenic regions of the FV3 genome. Our sequence analysis indicates that a fraction of these viral miRNAs is targeting the 3’-UTR regions of Xenopus genes involved in interferon (IFN)-dependent responses, including particularly those encoding IFN receptor subunits and IFN-regulatory factors (IRFs).ConclusionsUsing the FV3 model, this study provides a first genome-wide analysis of non-coding regulatory mechanisms adopted by ranaviruses to epigenetically regulate both viral and host gene expressions, which have co-evolved to interact especially with the host IFN response.

Coordinated expression of Jumonji and AHCY under OCT transcription factor control to regulate gene methylation in wood frogs during anoxia.

Wood frogs (Rana sylvatica) can survive extended periods of whole body freezing. Freezing imparts multiple stresses on cells that include anoxia and dehydration, but these can also be experienced as independent stresses. Under anoxia stress, energy metabolism is suppressed, and pro-survival pathways are prioritized to differentially regulate some transcription factors including OCT1 and OCT4. Jumonji C domain proteins (JMJD1A and JMJD2C) are hypoxia responsive demethylases whose expression is accelerated by OCT1 and OCT4 which act to demethylate genes related to the methionine cycle. The responses by these factors to 24h anoxia exposure and 4h aerobic recovery was analyzed in liver and skeletal muscle of wood frogs to assess their involvement in metabolic adaptation to oxygen limitation. Immunoblot results showed a decrease in JMJD1A levels under anoxia in liver and muscle, but an increase was observed in JMJD2C demethylase protein in anoxic skeletal muscle. Protein levels of adenosylhomocysteinase (AHCY) and methionine adenosyl transferase (MAT), enzymes of the methionine cycle, also showed an increase in the reoxygenated liver, whereas the levels decreased in muscle. A transcription factor ELISA showed a decrease in DNA binding by OCT1 in the reoxygenated liver and anoxic skeletal muscle, and transcript levels also showed tissue specific gene expression. The present study provides the first analysis of the role of the OCT1 transcription factor, associated proteins, and lysine demethylases in mediating responses to anoxia by wood frog tissues.

Heavy Metal Accumulation in the Tissues and Organs of Pelophylax ridibundus (Pallas, 1771) and Pelophylax lessonae (Camerano, 1882) (Amphibia: Ranidae) Living in the Waterbodies of Nizhniy Novgorod

A comprehensive atomic-emission study and comparative assessment were done of the content and distribution of heavy metals (Mn, Cu, Cr, Al, Fe, Zn, and Sr) in the organs and tissues (muscles, skin, bones, gonads, liver, heart, spleen, and blood) of marsh and pool frogs collected in the lakes of a big industrial city (Nizhni Novgorod). High concentrations of heavy metals were found in the spleen (Cr), bones (Zn and Sr), liver (Cu) of lake frogs. High coefficients of the biological absorption of Mn, Sr, Zn (bone tissue), Fe (liver, spleen), and Cu (heart) were found in pool frogs. The cumulative properties of heavy metals were estimated from the accumulation coefficient established. Species regularities of heavy metal accumulation from water are presented in descending order as the following series: Zn &gt; Cr &gt; Al &gt; Cu &gt; Fe &gt; Sr &gt; Mn for pool frogs; and Fe &gt; Zn &gt; Mn &gt; Cu &gt; Cr &gt; Al &gt; Sr for marsh frogs. The priority intake of chromium, manganese, aluminum and strontium from the aqueous medium into the body occurred through the skin. Active zinc accumulation occurred mainly through food, which was confirmed by high values of the biological absorption coefficient of zinc for the liver of Pelophylax ridibundus and Pelophylax lessonae. The dependence of the heavy metal accumulation in the muscles from the organs in contact with the external environment (skin) and the digestive system (liver) was established by regression analysis. The most important adaptation of tailless amphibians is their ability to prevent excessive accumulation of heavy metals in their body, while living in the conditions of increased environmental pollution. Our results obtained speak for the high accumulation of heavy metals in the body of tailless amphibians, determined by the conditions of the aquatic environment and the bioavailability of these metals.