Facilitate the Male Sexual Differentiation of Chicken Embryos.

BACKGROUND

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.

OBJECTIVE

To explore the function of screened by RNA-seq sequencing on sex differentiation during the development of chicken embryos.

METHODS

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.

RESULTS

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.

CONCLUSION

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.