部分去除原生殖细胞不影响红鲫的性别分化

王芳; 欧密; 张新铖; 刘海洋; 罗青; 费树站; 赵建; 李凯彬

部分去除原生殖细胞不影响红鲫的性别分化

Inducible Primordial Germ Cell Removal without Altering Sexuality of Red Crucian Carp

摘要

摘要:
目的获得针对红鲫原生殖细胞（Primordial germ cell，PGC）的可诱导去除品系，了解PGC删减对其性别发育和繁殖的影响，更好理解不同鱼类发育早期PGC数量与其性别发育的关系。
方法利用转基因方法构建PGC特异表达硝基还原酶的红鲫品系；甲硝唑（Metronidazole，Mtz）浸泡定向去除发育早期转基因鱼PGC，通过连续观察、定期采样、解剖、显微结构观察等方法研究PGC删减对个体性别发育的影响。
结果获得PGC标记特异荧光的稳定遗传红鲫品系，可通过荧光提示硝基还原酶（Nitroreductase, Ntr）在PGC中的表达；Mtz可被Ntr转化为具细胞毒性的代谢物，其对该品系红鲫早期胚胎的处理不仅能影响PGC的迁移，且特异地减少荧光标记细胞的数量，条件性诱导PGC去除；处理鱼养成后的性别比例和对照组相近，性腺未发现病变，且可正常繁殖。
结论发育早期的性细胞的减少并不能干扰红鲫性别发育，或红鲫性别分化依赖于性腺的体细胞。

Abstract:
Objective A primordial germ cell (PGC)-removed red crucian carp was obtained to investigate effects of the genetic alternation on the sexual development and reproduction of the fish.
Method A PGC-specific expression of nitroreductase (Ntr)-containing strain of red crucian carp line with PGC removed by a metronidazole (Mtz) immersion-targeted treatment at early development stage was continuously observed, sampled, and dissected for anatomical differentiations under a microscope.
Result A stable and heritable red crucian carp line carrying the specific PGC marker fluorescence was successfully secured. Since Mtz can be converted by Ntr into a cytotoxic metabolite, the induction in the fish did not affect the PGC migration nor the number of fluorescently labeled cells. Thus, the PGC-removed transgenic line showed no pathological changes in the gonads and continued to reproduce offsprings with a sex ratio similar to that of control.
Conclusion The removal of PGCs in red crucian carp at the embryonic stage exhibited no interference to the sexual development of the fish, and the sex differentiation of this fish may depends on the somatic cells of gonads.

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