Abstract
To elucidate the role of anti-müllerian hormone (AMH) in regulating the development of ovarian follicles in laying hens, the expressions of follicle-stimulating hormone receptor (FSHR), AMH receptor type 2 (AMHR2), steroidogenic-related genes steroidogenic acute regulatory protein (STAR), cytochrome P450 side-chain cleavage (CYP11A1), and 3β-hydroxysteroid dehydrogenase (HSD3B1) genes were measured from different sized follicles and granulosa cells. The results showed that the expressions of FSHR and AMHR2 genes were higher in small follicles and decreased after follicular selection. Oppositely, the expressions of STAR, CYP11A1, and HSD3B1 were significantly increased after follicular selection. It indicated that AMHR2 might mediate AMH suppression in the stimulating effects of follicle-stimulating hormone (FSH) on steroidogenic-related genes expression. To make sure the effects of AMH in this process, a total of 40 hens were treated (negative control, sham operation, 150 ng AMH/d or 300 ng AMH/d) for 25 d. We analyzed ovarian morphology, progesterone concentration in blood plasma, and the expressions of steroidogenic genes in ovaries and follicles. The AMH300 group had significantly lower weight of ovary and hierarchical follicles. Egg weight and ovary weight in AMH150 group were higher than those of sham operation and AMH300 groups, so did hierarchical follicles weight. The steroidogenic genes expressions showed an increase in ovarian tissue and the largest follicle of AMH150 and AMH300 groups. However, progesterone level in the blood was reduced by AMH injection with different concentrations. To further verify the above results, granulosa cells from 6 to 8 mm follicles were cultured with AMH (0, 5, 10, 20, 40, or 80 ng/mL). The results revealed that excessive AMH (80 ng/mL) exerted an inhibitory effect on progesterone synthesis and the expressions of STAR, CYP11A1, and HSD3B1. However, these genes expressions showed a significant increase in 20 ng/mL AMH-treated group. In summary, AMH inhibited the development of prehierarchical follicles in laying hens. The effects of AMH treatment with different concentrations on follicle development showed the follicle was selected by changing FSH responsiveness of prehierarchical follicles.