Abstract

Mono-Sex culturing is an important methodology for intensive livestock and poultry production. Here, Hintw was identified as a potential key gene in sex-determination process in chickens via RNA-seq. Then we developed an effective method to interfere or overexpress Hintw in chicken embryos through the intravascular injection. QRT-PCR, ELISA and H&E staining were used to detect the effects of Hintw on gonadal development of chicken embryos. Results showed that Hintw exhibited a female-biased expression pattern in the early stage of PGCs (primordial germ cells) in embryonic gonads. The qRT-PCR analysis showed that Foxl2, Cyp19a1 in females were upregulated under the overexpression of Hintw, while Sox9 and Dmrt1 were downregulated Hintw. Overexpression of Hintw can promote the development of gonadal cortex, while interference with Hintw show the opposite result. Additionally, we found that overexpression of the Hintw in male chicken embryos could inhibit androgen levels and increase estrogen levels. On the other hand, interfering with Hintw in female chicken embryos decreased estrogen levels and increased androgen levels. In conclusion, this work sets the basis for the understanding of the molecular regulatory network for the sex-determination process in chicken embryos as well as providing the theoretical basis for mono-sex culturing of poultry.

摘要

单性养殖是畜禽集约化生产的重要方法。在这里，通过 RNA-seq， Hintw被确定为鸡性别决定过程中潜在的关键基因。然后我们开发了一种有效的方法，通过血管内注射在鸡胚胎中干扰或过表达Hintw 。采用QRT-PCR、ELISA和H&E染色检测Hintw对鸡胚胎性腺发育的影响。结果表明，Hintw在胚胎性腺的 PGC（原始生殖细胞）早期表现出女性偏向的表达模式。qRT-PCR分析表明，雌性Foxl2、Cyp19a1在过表达下表达上调Hintw，而Sox9和Dmrt1被下调Hintw。过表达Hintw可促进性腺皮质发育，干扰Hintw则相反。此外，我们发现雄性鸡胚胎中Hintw的过度表达可以抑制雄激素水平并增加雌激素水平。另一方面，干扰Hintw在雌性鸡胚胎中，雌激素水平降低，雄激素水平升高。总之，这项工作为理解鸡胚胎性别决定过程的分子调控网络奠定了基础，并为家禽的单性培养提供了理论基础。