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王金霓, 吴俣, 王子嘉, 黄安, 楼灵媛, 庄天依, 孙伟, 钱国英, 郭银, 葛楚天. 中华鳖生殖细胞对性腺分化的影响[J]. 水生生物学报, 2023, 47(11): 1849-1857. DOI: 10.7541/2023.2023.0073
引用本文: 王金霓, 吴俣, 王子嘉, 黄安, 楼灵媛, 庄天依, 孙伟, 钱国英, 郭银, 葛楚天. 中华鳖生殖细胞对性腺分化的影响[J]. 水生生物学报, 2023, 47(11): 1849-1857. DOI: 10.7541/2023.2023.0073
WANG Jin-Ni, WU Yu, WANG Zi-Jia, HUANG An, LOU Ling-Yuan, ZHUANG Tian-Yi, SUN Wei, QIAN Guo-Ying, GUO Yin, GE Chu-Tian. ELIMINATION OF GERM CELLS ON GONADAL DIFFERENTIATION IN PELODICUS SINENSIS[J]. ACTA HYDROBIOLOGICA SINICA, 2023, 47(11): 1849-1857. DOI: 10.7541/2023.2023.0073
Citation: WANG Jin-Ni, WU Yu, WANG Zi-Jia, HUANG An, LOU Ling-Yuan, ZHUANG Tian-Yi, SUN Wei, QIAN Guo-Ying, GUO Yin, GE Chu-Tian. ELIMINATION OF GERM CELLS ON GONADAL DIFFERENTIATION IN PELODICUS SINENSIS[J]. ACTA HYDROBIOLOGICA SINICA, 2023, 47(11): 1849-1857. DOI: 10.7541/2023.2023.0073

中华鳖生殖细胞对性腺分化的影响

ELIMINATION OF GERM CELLS ON GONADAL DIFFERENTIATION IN PELODICUS SINENSIS

  • 摘要: 为了探究爬行动物生殖细胞如何影响体细胞及生殖细胞数量是否会影响性别决定和分化, 研究通过向中华鳖早期胚胎注射白消安消除了内源生殖细胞。结果显示: 在白消安处理后, ZW和ZZ性腺生殖细胞数量急剧下调, 几乎消失, 但性腺形态没有发生明显改变, ZW性腺仍旧呈现雌性状态并表达雌性标志蛋白FOXL2和CYP19A1, ZZ性腺仍旧呈现雄性状态并表达雄性标志蛋白DMRT1和SOX9。研究表明在爬行动物中消除生殖细胞不影响性别分化。

     

    Abstract: In reptiles, primordial germ cells proliferate massively in the female cortex and entry the “mitotic arrest” in the male medulla when they reach the genital ridge. Somatic cells facilitate germ cells development and differentiation. However, it is not clear how germ cells affect somatic cells and whether germ cells depletion could influence the sex differentiation. In order to investigate whether germ cells have impacts on somatic cells and gonadal sex differentiation, we depleted the endogenous germ cells in ZZ and ZW genetic Pelodicus sinensis by injecting of 20 mg/kg busulfan into the embryos in this study. Immunofluorescence of VASA showed that the germ cells are depleted after treatment of busulfan in both ZZ and ZW gonads. H & E staining analysis showed that the morphology of cortex and medullary cords had no difference with or without busulfan treatment in ZZ and ZW turtles. The busulfan treated ZW gonads expressed the ovary-specific marker FOXL2 and CYP19A1, while the busulfan treated ZZ gonads showed immunofluorescence staining of testicular marker protein DMRT1 and SOX9. The Real-time PCR analysis showed that the busulfan-treated ZW gonads still expressed high levels of Cyp19a1 and Foxl2 and low levels of Dmrt1 and Sox9, while the busulfan-treated ZZ gonads expressed high levels of Dmrt1 and Sox9 and relative low levels of Cyp19a1 and Foxl2. These results suggest that the gonads sex do not change after the depletion of germ cells in both ZW and ZZ genetic turtles. In conclusion, our study first proved that the germ cells depletion do not alter the sex differentiation in GSD reptiles, which can provide reference significance for breeding of Pelodicus sinensis.

     

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