Testicular Transcriptome of Males and Pseudo-Males Provides Important New Insight into Sex Reversal of Rana dybowskii

SIMPLE SUMMARY: Aquaculture in Northeast China has favored Rana dybowskii (R. dybowskii) because of its regional specificity and the economic value of the female frog’s ovary, and the increasing proportion of female frogs is beneficial to improve the economic efficiency of aquaculture. Unfortunately, the sexual reversal from genotypic female to phenotypic male has led to a decrease in the proportion of females in the breeding population. In this study, differentially expressed genes between testes of sex reversal and male testes were screened by transcriptome sequencing. These findings provide a comprehensive analysis of gene expression in R. dybowskii sex reversal from genotypic female to phenotypic male and provide new ideas for sex control of R. dybowskii. ABSTRACT: Rana dybowskii (R. dybowskii) is an ecological species found in China, Japan, Korea, and Russia. Like most amphibians, R. dybowskii lacks heterotypic sex chromosomes, limiting the in-depth study of sex determination and sex reversal mechanisms. Previous studies have shown that certain environmental factors can modify R. dybowskii genotypic females into phenotypic males, but the mechanism is still unknown. Considering the difficulties in identifying and collecting sex reversal gonads at different stages of differentiation under natural conditions, testes from sexually mature wild adult R. dybowskii were taken in this study, and the genotypic sex of individuals and sex reversal were identified by two male-linked genetic markers reported in our most recent findings. Transcriptome sequencing was performed on testicular tissue from males and pseudo-males, as well as female ovary tissue. The results show that the gene expression patterns of pseudo-males’ testes were similar to those of the males but highly differed from females’ ovaries. One hundred and seventeen differentially expressed genes between testes of pseudo-males and males were found, and the up-regulation of doublesex and mab-3 related transcription factor 1 (Dmrt1) in testes of pseudo-males may play a key role in R. dybowskii sex reversal.