Abstract
Cytochrome P450 Family 19 SubFamily A member 1 (CYP19A1) gene encodes an aromatase which regulates the sexual differentiation in vertebrates by initiating and maintaining 17β-Estradiol (E2) synthesis. Here, we described the spatiotemporal expression pattern of CYP19A1 and its functional role in the embryonic gonad development in amphoteric chickens (Gallus gallus). Results showed that CYP19A1 exhibited a sexually dimorphic expression pattern in female gonads early at embryonic day 5.5 (HH 28) and robustly expressed within the cytoplasm in ovarian medullas. Most importantly, we induced the gonadal sex reversal by ectopically delivering the aromatase inhibitor (AI) or estradiol (E2) into chicken embryos. To further explore the role of CYP19A1 in chicken embryonic sexual differentiation, we successfully developed an effective method to deliver lentiviral particles with CYP19A1 manipulation into chicken embryos via embryonic intravascular injection. The analysis of interference and overexpression of CYP19A1 provided solid evidences that CYP19A1 is both necessary and sufficient to initiate sex differentiation toward female in chicken embryos. Collectively, this work demonstrates that CYP19A1 is a crucial sex differentiation gene in the embryonic development, which provides a foundation for understanding the mechanism of sex determination and differentiation in chickens.
Highlights
In mammals, the well-developed system for single-sex reproduction is based on the known sex-determination system [1,2]
We described the expression pattern of CYP19A1 and its functional role in chicken embryonic gonads development by mediating the estriol level via aromatase inhibitor (AI) treatment and manipulating CYP19A1 using lentivirus-mediated RNAi and overexpressing systems
The results showed that the CYP19A1 transcript was significantly highly expressed in female gonads throughout the period of sex determination to born, from as early as day 5.5 (HH 28) to the day 21.5, but barely expressed in male gonad (Figure 1A)
Summary
The well-developed system for single-sex reproduction is based on the known sex-determination system [1,2]. As egg-laying chickens need the female offspring while broiler chickens need males, the specific sex selection helps to reduce the cost and promote the poultry industry development [5]. It is essential to identify and characterize the function of critical sex differentiation genes for improving the single-sex reproduction. Cytochrome P450 Family 19 SubFamily A member 1 (CYP19A1), a vital enzyme for estrogen (17β-Estradiol, E2) synthesis in most vertebrates, modulates steroid hormones involved in the sex differentiation among amphibians, fishes, reptiles, birds and mammals [7,8,9,10].
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