Evolution of sexual dimorphism in the plateau brown frog fails to obey Rensch’s rule

Rensch’s rule describes sexual size dimorphism (SSD) that decreases with increasing body size when females are larger than males and SSD that increases when males are larger than females. The plateau brown frog Rana kukunoris, a species endemic to the eastern Tibetan Plateau, exhibits female-biased size dimorphism. Using data on body size from 26 populations and age from 21 populations, we demonstrated that SSD did not increase with increasing mean female snout-vent length (SVL) when controlling for sex-specific age structure, failing to support the Rensch’s rule. Thus, we suggest that fecundity selection (favouring large female size) balances out sexual selection (favouring large male size), which results in a similar divergence between males and females body size. In addition, sex-specific age differences explained most of the variation of SSD across populations.

Ecogeographical Adaptation Revisited: Morphological Variations in the Plateau Brown Frog along an Elevation Gradient on the Qinghai–Tibetan Plateau

Several anurans have broad elevational and latitudinal distribution ranges; distinct species and populations may face various environmental and selection stresses. Due to their environmental sensitivity, adaptation is critical for the long-term persistence of anurans. Previous studies have tried to identify the ecogeographical pattern and its mechanism in anurans, suggesting different patterns, but the related explanatory mechanisms are yet to be generally supported and are suggested to be complicated. To explore the elusive mechanisms, we studied the morphological variation of the plateau brown frog (Rana kukunoris) along an elevational gradient on the eastern margin of the Qinghai–Tibetan Plateau. Using body size, extremity length, and the ratio between them (extremities/body size) as testing indicators, we examined potential ecogeographical adaptations and investigated how environmental and biological factors could shape the morphological development in R. kukunoris. We found that males and females showed different variations in body size and extremities along the elevational gradient, whereas both of them showed a decreasing extremities/body size ratio along elevation. Together with the strong correlations between environmental and biological factors and the morphometrics, we identified ecogeographical adaptation and a sexual difference in the selective pressures on the extremities and body size of the plateau brown frog. Our results imply that geographic variations in anuran morphological traits should be understood as an outcome of environmental and biological factors. Furthermore, ecogeographical adaptation in anurans can manifest as an interactive change between body size and extremities.

Expression of Glycogenic Genes in the Oviduct of Chinese Brown Frog (Rana Dybowskii) During Pre-Hibernation

Background: The oviduct of Chinese brown frog (Rana dybowskii) expands specifically during pre-hibernation. However, the underlying molecular mechanism remains unclear. Results: This study investigated the mRNA and protein expression levels of glycogenic genes in the oviduct of Rana dybowskii during the breeding season and pre-hibernation. Morphological studies revealed increased weight and enlarged diameter of the oviduct in pre-hibernation. Furthermore, significantly increased glycogen level was detected in the oviduct during pre-hibernation when compared to the breeding season. Transcriptome analysis further identified that the differentially expressed genes in the synthesis and metabolism pathways of carbohydrates in the oviduct during pre-hibernation. The relative mRNA levels of glycolysis and glycogenesis-related genes were up-regulated, while gluconeogenesis-related genes were down-regulated during pre-hibernation. Also, Western blot showed that glucose transporter and glycogen synthesis-related proteins, such as total-GYS, and p-GSK-3β were highly expressed in the oviduct during pre-hibernation. In addition, immunohistochemical data showed that glucose transporter and total-GYS, p-GYS, total-GSK-3β and p-GSK-3β related to glycogen synthesis were both expressed regionally in the oviduct of Rana dybowskii during the breeding season and pre-hibernation. Conclusions: The data suggests that carbohydrate metabolism plays an important role in the hibernation process of Rana dybowskii, and provides a new insight into the mechanism underlying the expansion of the oviduct of Rana dybowskii during pre-hibernation.

Immunoreactivities of AR, ERα, ERβ and aromatase in the nuptial pad of Chinese brown frog (Rana dybowskii) during pre-hibernation and the breeding period

There is a prominent local raised pad called nuptial pad on the forelimb of Chinese brown frog (Rana dybowskii), which is hypothetically concluded as an enhancement of the grip and a spreader of pheromone during the amplexus. In this study, we investigated the immunolocalization and protein expression levels of AR, ERα, ERβ and aromatase in the nuptial pad of R. dybowskii during pre-hibernation and the breeding period. Histologically, the annual development of the nuptial pad in R. dybowskii is manifested as the larger area of specialized mucous gland and the longer length of papillary epidermal projection during the breeding period. AR, ERα, ERβ and aromatase are present in the stratum granulosum, stratum spinosum, stratum basale and the secretory portion of specialized mucous glands during both periods. Western blotting results confirmed that AR, ERα and ERβ protein levels are higher during pre-hibernation than those during the breeding season. These results suggest that nuptial pad is the direct target organ of androgen and estrogen. Androgen may participate in the regulation of annual development and glandular function of nuptial pad, and estrogen may play an endocrine, autocrine or paracrine role during pre-hibernation and the breeding period.

Evolution of Sex Chromosome Heteromorphy in Geographic Populations of the Japanese Tago’s Brown Frog Complex

The sex chromosomes of most anuran amphibians are characterized by homomorphy in both sexes, and evolution to heteromorphy rarely occurs at the species or geographic population level. Here, we report sex chromosome heteromorphy in geographic populations of the Japanese Tago’s brown frog complex (2n = 26), comprising Rana sakuraii and R. tagoi. The sex chromosomes of R. sakuraii from the populations in western Japan were homomorphic in both sexes, whereas chromosome 7 from the populations in eastern Japan were heteromorphic in males. Chromosome 7 of R. tagoi, which is distributed close to R. sakuraii in eastern Japan, was highly similar in morphology to the Y chromosome of R. sakuraii. Based on this and on mitochondrial gene sequence analysis, we hypothesize that in the R. sakuraii populations from eastern Japan the XY heteromorphic sex chromosome system was established by the introduction of chromosome 7 from R. tagoi via interspecies hybridization. In contrast, chromosome 13 of R. tagoi from the 2 large islands in western Japan, Shikoku and Kyushu, showed a heteromorphic pattern of constitutive heterochromatin distribution in males, while this pattern was homomorphic in females. Our study reveals that sex chromosome heteromorphy evolved independently at the geographic lineage level in this species complex.

Evolution of a Multiple Sex-Chromosome System by Three-Sequential Translocations among Potential Sex-Chromosomes in the Taiwanese Frog Odorrana swinhoana

Translocation between sex-chromosomes and autosomes generates multiple sex-chromosome systems. It happens unexpectedly, and therefore, the evolutionary meaning is not clear. The current study shows a multiple sex chromosome system comprising three different chromosome pairs in a Taiwanese brown frog (Odorrana swinhoana). The male-specific three translocations created a system of six sex-chromosomes, ♂X1Y1X2Y2X3Y3 -♀X1X1X2X2X3X3. It is unique in that the translocations occurred among three out of the six members of potential sex-determining chromosomes, which are known to be involved in sex-chromosome turnover in frogs, and the two out of three include orthologs of the sex-determining genes in mammals, birds and fishes. This rare case suggests sex-specific, nonrandom translocations and thus provides a new viewpoint for the evolutionary meaning of the multiple sex chromosome system.

First detection of ranavirus in a wild population of Dybowski’s brown frog (Rana dybowskii) in South Korea

Background Ranavirus is an emerging infectious disease which has been linked to mass mortality events in various amphibian species. In this study, we document the first mass mortality event of an adult population of Dybowski’s brown frogs (Rana dybowskii), in 2017, within a mountain valley in South Korea. Results We confirmed the presence of ranavirus from all collected frogs (n = 22) via PCR and obtained the 500 bp major capsid protein (MCP) sequence from 13 individuals. The identified MCP sequence highly resembled Frog virus 3 (FV3) and was the same haplotype of a previously identified viral sequence collected from Huanren brown frog (R. huanrenensis) tadpoles in South Korea. Human habitat alteration, by recent erosion control works, may be partially responsible for this mass mortality event. Conclusion We document the first mass mortality event in a wild Korean population of R. dybowskii. We also suggest, to determine if ranavirus infection is a threat to amphibians, government officials and researchers should develop continuous, country-wide, ranavirus monitoring programs of Korean amphibian populations.