Page 68 - 《水产学报》2025年第6期

梁肖，等 水产学报, 2025, 49(6): 069605

The role of sox9 gene in male gonadal differentiation of
Pelodiscus sinensis

1,2
3
1,2
2,3
LIANG Xiao , JIN Lin , LI Pan , YANG Bangsai ,
2,3
2,3*
QIAN Guoying , GE Chutian , SUN Wei 2,3*
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;
2. Institute of Animal Sex and Development, Zhejiang Wanli University, Ningbo 315100, China;
3. College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China
Abstract: Chinese soft-shelled turtles (Pelodiscus sinensis) possess apparent sexual dimorphism on production traits, and the
realization of all-male breeding is the important measure for the improvement of the industry. In this study, the highly con-
served gene sox9 in vertebrate male differentiation was chosen as manipulating object, and by injecting Lentivirus-sox9-shRNA
interference and overexpression vector system into embryos before sexual differentiation, we discussed the specific function of
sox9 during the early testicular differentiation of P. sinensis from the level of histology and molecular study. The results of the
loss-of-function experiments showed that the appearance and internal structure of ZZ embryonic gonads with sox9 knockdown
underwent significant feminization, and the germ cells presented female distribution pattern. The mRNA expression of male
specific gene dmrt1 and amh significantly decreased, and that of female specific gene foxl2 obviously increased. Besides, the
expressive signals of the DMRT1 protein almost disappeared, while FOXL2 protein was activated and abundantly expressed,
indicating complete sex reversal from male to female on gonads(ratio of reversal: 87.9%). The results of the gain-of-function
experiments showed that 42.5% of ZW embryos overexpressing sox9 differentiated towards male. The expression of dmrt1 and
amh were upregulated while foxl2 was downregulated, and the fluorescent signals of both DMRT1 and FOXL2 were detected
simultaneously in the same gonad, indicating incomplete sex reversal. These findings above demonstrate that sox9 is the neces-
sary gene for early testicular formation in P. sinensis, and it does participate in the process of regulating male differentiation,
which establishes theoretical foundation for the following analysis of sexual differentiation mechanism of P. sinensis and
provides breeding target for realizing the sex control of P. sinensis at the same time.
Key words: Pelodiscus sinensis; sox9 gene; sexual differentiation; sex reversal; male
Corresponding authors: GE Chutian. E-mail: cge@zwu.edu.cn;
SUN Wei. E-mail: sunwei0802@zwu.edu.cn
Funding projects: National Natural Science Foundation of China (32325049, U22A20529); Natural Science Foundation of
Ningbo (2022J193)

https://www.china-fishery.cn 中国水产学会主办 sponsored by China Society of Fisheries
12