Chen Chen, Zhou Shujian, Lian Ziyi, Jiang Jingyi, Gao Xiaomin, Hu Cai, Zuo Qisheng, Zhang Yani, Chen Guohong, Jin Kai, Li Bichun
Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China.
Front Physiol. 2022 Apr 7;13:856980. doi: 10.3389/fphys.2022.856980. eCollection 2022.
Sex differentiation is a complex and precisely regulated process by multiple genes in chicken. However, it is still unclear on the key genes of sex differentiation.
To explore the function of screened by RNA-seq sequencing on sex differentiation during the development of chicken embryos.
was differentially expressed from the RNA-seq of ESCs and PGCs in male and female chickens. Then, we established an effective method to overexpression or knocking down the expression of in ovo and , respectively. Histomorphological observation, qRT-PCR and ELISA were applied to detect the function of in the process of male sex differentiation by injecting vectors into embryos at day 0.
It showed that has significant male preference in embryonic day 4.5, such phenomenon persisted during the growth period of chicken embryos. Morphological observation results showed that the gonads on both sides of genetic male (ZZ) embryos with knocking down developed asymmetrically, the gonadal cortex became thicker showing the typical characteristics of genetic female (ZW) gonads. Furthermore, the expression of Cyp19a1, which dominates female differentiation, was significantly increased, while the expression of male marker genes Dmrt1, Sox9, WT1 and AR was significantly downregulated. In addition, the concentration of testosterone also significantly decreased, which was positively correlated with the expression of ( < 0.01). Conversely, the ZW embryo showed defeminized development when was overexpressed.
We prove that the is a novel gene through the male sexual differentiation gene regulation process and synthesis of testosterone, which construct the basis for understanding the molecular mechanism of sex differentiation in chickens.
性别分化是鸡体内由多个基因参与的复杂且精确调控的过程。然而，性别分化的关键基因仍不清楚。
探究通过RNA-seq测序筛选出的基因在鸡胚发育过程中对性别分化的作用。
该基因在雌雄鸡胚胎干细胞和原始生殖细胞的RNA-seq中差异表达。然后，我们分别建立了在鸡胚内过表达或敲低该基因表达的有效方法。通过在胚胎孵化第0天注射载体，应用组织形态学观察、qRT-PCR和ELISA检测该基因在雄性性别分化过程中的作用。
结果显示，该基因在胚胎发育第4.5天具有明显的雄性偏好，这种现象在鸡胚生长期间持续存在。形态学观察结果表明，敲低该基因的遗传雄性（ZZ）胚胎两侧性腺发育不对称，性腺皮质增厚，呈现出遗传雌性（ZW）性腺的典型特征。此外，主导雌性分化的Cyp19a1表达显著增加，而雄性标记基因Dmrt1、Sox9、WT1和AR的表达显著下调。此外，睾酮浓度也显著降低，且与该基因的表达呈正相关（P<0.01）。相反，过表达该基因时，ZW胚胎出现去雌性化发育。
我们通过雄性性别分化基因调控过程和睾酮合成证明该基因为新基因，这为理解鸡性别分化的分子机制奠定了基础。
