斑马鱼ppp6r3调控性腺分化和配子发生的作用研究

doi:10.16288/j.yczz.25-093

遗传 ›› 2025, Vol. 47 ›› Issue (9): 1023-1031.doi: 10.16288/j.yczz.25-093

斑马鱼ppp6r3调控性腺分化和配子发生的作用研究

高云海¹^,²(), 邓家杰², 肖霄², 潘鲁湲³, 何牡丹²(), 张蕴斌¹()

1.中国科学院分子细胞科学卓越创新中心，上海 200031
2.中国科学院水生生物研究所，武汉 430070
3.中国科学院水生生物研究所，国家水生生物种质资源库国家斑马鱼资源中心，武汉 430070

收稿日期:2025-04-02 修回日期:2025-04-24 出版日期:2025-05-13 发布日期:2025-05-13
通讯作者: 张蕴斌，博士，高级工程师，研究方向：模式动物。E-mail: zhangyb@sibcb.ac.cn;
何牡丹，博士，副研究员，研究方向：鱼类遗传育种。E-mail: hemudan@ihb.ac.cn
作者简介:高云海，硕士研究生，专业方向：生物与医药。E-mail: gaoyunhai2022@sibcb.ac.cn
基金资助:
国家重点研发计划(2022YFF0710500);国家重点研发计划(2024YFD2402300);科技创新2030重大项目(2023ZD04069)

Role of ppp6r3 in zebrafish gonadal differentiation and gametogenesis

Yunhai Gao¹^,²(), Jiajie Deng², Xiao Xiao², Luyuan Pan³, Mudan He²(), Yunbin Zhang¹()

1. Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China
2. Institute of Hydrobiology, Chinese Academy of Science, Wuhan 430070, China
3. China Zebrafish Resource Center, National Aquatic Biological Resource Center, Institute of Hydrobiology, Chinese Academy of Science, Wuhan 430070, China

Received:2025-04-02 Revised:2025-04-24 Published:2025-05-13 Online:2025-05-13
Supported by:
National Key R&D Program of China(2022YFF0710500);National Key R&D Program of China(2024YFD2402300);STI 2030-Major Projects(2023ZD04069)

摘要/Abstract

摘要：

鱼类的性腺分化与配子发生受到多种因子的调控。蛋白磷酸酶(protein phosphatase，PP)具有催化生物体中蛋白去磷酸化的功能。蛋白磷酸酶6 (protein phosphatase type 6，PP6)作为蛋白磷酸酶家族的一员，在性腺发育及配子发生过程中发挥重要功能。ppp6r3编码蛋白磷酸酶6调节亚基3，其在性腺分化和配子发生方面的作用还不清楚。本研究通过CRISPR/Cas9技术构建了两个斑马鱼ppp6r3突变品系，发现ppp6r3缺失后导致斑马鱼仅发育为雄鱼，且该突变体不能使野生型卵子受精。进一步研究发现，ppp6r3突变体精巢中精母细胞向成熟精子转换失败，精子发生受阻。综上所述，本研究构建了ppp6r3突变的斑马鱼模型，表现出性腺分化和配子发生的缺陷，为后续研究Ppp6r3调控生殖细胞增殖、分化及性别决定的机制提供了模型和参考。

关键词: Ppp6r3, 斑马鱼, 性腺分化, 配子发生

Abstract:

The gonad differentiation and gametogenesis of fish is regulated by various factors. Protein phosphatase (PP) have the function of catalyzing the dephosphorylation of proteins in organisms. As a member of the protein phosphatase family, protein phosphatase type 6 (PP6) plays an important role in gonadal development and gametogenesis. The role of ppp6r3, which encodes the regulatory subunit 3 of protein phosphatase 6, in gonadal differentiation and gametogenesis is still unclear. In this study, two zebrafish ppp6r3 mutant lines were constructed using CRISPR/Cas9 technology. It was found that the absence of ppp6r3 leads to the development of only male zebrafish, and these mutants are incapable of fertilizing wild-type eggs. Further investigations revealed that in the testes of ppp6r3 mutants, the transition of spermatocytes to mature sperm was blocked, disrupting spermatogenesis. In summary, this study established a ppp6r3 mutant model, exhibiting defects in gonadal differentiation and gametogenesis, which provides a new model for further research on the mechanisms by which Ppp6r3 regulates germ cell proliferation, differentiation, and sex determination.

Key words: Ppp6r3, zebrafish, gonadal differentiation, gametogenesis

高云海, 邓家杰, 肖霄, 潘鲁湲, 何牡丹, 张蕴斌. 斑马鱼ppp6r3调控性腺分化和配子发生的作用研究[J]. 遗传, 2025, 47(9): 1023-1031.

Yunhai Gao, Jiajie Deng, Xiao Xiao, Luyuan Pan, Mudan He, Yunbin Zhang. Role of ppp6r3 in zebrafish gonadal differentiation and gametogenesis[J]. Hereditas(Beijing), 2025, 47(9): 1023-1031.

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参考文献 19

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引物	序列(5′→3′)	用途
ppp6r3-gRNA-F	TGTAATACGACTCACTATAACGGATGGTTCATTCCAGGGGTTTTA GAGCTAGAAATAGC	gRNA扩增
gRNA-RP	AAAAAAAGCACCGACTCGGTGCCAC	gRNA扩增
ppp6r3-cas9-F	TGGAGGGATTAAGGTTGAGCT	靶点扩增和突变体筛选
ppp6r3-cas9-R	GTCCAAGACTCTGCCAGGTT	靶点扩增和突变体筛选
Ppp6r3-P1	CCTCGTAAATCCTCATCAAAAGCTGTAGAGTTTCATCAGTAAACT	单分子荧光检测
Ppp6r3-P2	GGTTGAGCGGAGGCTCGTTCTGTAAAAATCATCCAGTAAACCGCC
Ppp6r3-P3	CCTCGTAAATCCTCATCAAATCAAGACTTTGCTGAAGAAACTGGC
Ppp6r3-P4	GCTCTGGTTTTCTCCCAATAAGTATAAATCATCCAGTAAACCGCC
Ppp6r3-P5	CCTCGTAAATCCTCATCAAAAGTCTTCCCTCTTCCTGAGGAACTC
Ppp6r3-P6	CTATGTGTTTGATCATCAGATCCACAAATCATCCAGTAAACCGCC
Ppp6r3-P7	CCTCGTAAATCCTCATCAAATCCTGAGCAGGAGGTCCATAATAGC
Ppp6r3-P8	GCTGCTGTGGCTCAATGCAGGTGAGAAATCATCCAGTAAACCGCC
Ppp6r3-P9	CCTCGTAAATCCTCATCAAACTTCATTTAGCCAATTTAATACATC
Ppp6r3-P10	TATCAACCAGCCGCTGTATAACTTTAAATCATCCAGTAAACCGCC

斑马鱼ppp6r3调控性腺分化和配子发生的作用研究

Role of ppp6r3 in zebrafish gonadal differentiation and gametogenesis

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