Expression Characteristics Analysis of srsf3 Gene in Tissues and Gonad During Development Stages of Chinese Giant Salamander (Andrias davidianus)
MENG Fang1,2, FANG Jun-Chao2, TIAN Hai-Feng2, SHAO Peng1*, HU Qiao-Mu2*
1 College of Aquaculture, Tianjin Agricultural University, Tianjin 300380, China; 2 Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China
Abstract：There is a significant difference in the growth of male and female giant salamanders (Andrias davidianus), and it has been confirmed that the serine/arginine-rich splicing factor 3 (srsf3) gene is involved in the selective splicing of nuclear receptor subfamily 5 group A member 2 (Nr5a2), a gene related to sex differentiation in giant salamanders. To investigate the mechanism of srsf3 gene in sex differentiation of Chinese giant salamander , qPCR technology was used to analyze the changes in expression levels in different tissues, gonads at different developmental stages, and gonads induced by sex hormones; Fluorescence in situ hybridization (FISH) was used to determine the expression and localization of the srsf3 gene in the testes and ovaries of giant salamanders. Based on the analysis of the transcriptome sequencing results of the giant salamander gonads, the full-length of srsf3 gene was 1 478 bp, with an open reading frame of 504 bp, encoding 167 amino acids. Systematic evolutionary analysis showed that the amino acid sequence of the giant salamander SRSF3 was closely related to the Xenopus tropicalis. The qPCR results showed that the expression level of srsf3 gene was highest in the ovaries of giant salamanders, followed by pituitary tissue, slightly lower in muscle and heart tissues, and low expression in liver, spleen, lung, and stomach tissues. The expression of the srsf3 gene in the gonads of giant salamanders at different stages showed that the ovaries were significantly higher than the testes at ages 2~5 (P<0.05); The expression level of srsf3 gene reached its highest in the ovaries at the age of 2, and then decreased year by year in the ovaries. The expression level tended to stabilize in the ovaries at the age of 4-6. The expression level of the srsf3 gene was highest in sex reversed males, followed by sex reversed females; The expression level of srsf3 gene in the sex reversal ovaries was significantly higher than that in the ovaries (P<0.05), and the expression level in the sex reversal testes was significantly higher than that in the testes (P<0.05). The FISH results showed that the srsf3 gene was expressed in both the ovaries and testes of the giant salamander. The above studies indicated that the srsf3 gene was involved in the gonadal development of giant salamanders and plays an important role in the process of sex reversal in giant salamanders. Functional analysis of the srsf3 gene in this study helps to deepen the understanding of the mechanism of sex differentiation in amphibians.
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