Abstract

Goose (Anser cygnoides) are highly broody with low egg production, and large number of granulosa cells undergo apoptosis during broodiness. Our previous study has found that miR-34c-5p was highly abundant in the ovary of goose with broodiness phenotype. However, the mechanism that miR-34c-5p regulates granulosa cells function remains unclear. Here, we demonstrated that broody goose had higher levels of miR-34c-5p than that in laying goose by qRT-PCR. The dual luciferase reporter assay showed that Bcl2 was identified as a direct target of miR-34c-5p, which could be negatively regulated by miR-34c-5p. Furthermore, over-expression of miR-34c-5p significantly increased the rate of apoptosis and slowed down the proliferation of granulosa cells by inhibiting the Bcl2 expression, whereas the opposite trend was obtained when granulosa cells were supplemented with miR-34c-5p inhibitors. In addition, Bcl2 mRNA level was lower in goose with a brooding phenotype than that in goose with an egg-laying phenotype. Taken together, the data suggested that miR-34c-5p regulated granulosa cells apoptosis and brooding behavior by targeting Bcl2, which not only contribute to reveal the potential mechanism of miR-34c-5p underlying granulosa cells apoptosis in goose, but also provides an effective strategy to reduce the incidence of broodiness and improve the egg production.

摘要

鹅( Anser cygnoides)多育性低产蛋率，大量颗粒细胞在育雏过程中发生凋亡。我们之前的研究发现miR-34c-5p在具有抱窝表型的鹅的卵巢中含量很高。然而，miR-34c-5p调节颗粒细胞功能的机制仍不清楚。在这里，我们通过 qRT-PCR证明了孵蛋鹅的miR-34c-5p水平高于下蛋鹅。双荧光素酶报告基因检测表明，Bcl2被鉴定为miR-34c-5p的直接靶标，可被miR-34c-5p负调控。此外，过表达miR-34c-5p通过抑制Bcl2表达显着增加细胞凋亡率并减缓颗粒细胞的增殖，而当颗粒细胞补充miR-34c-5p抑制剂时获得相反的趋势。此外，具有育雏表型的鹅的Bcl2 mRNA 水平低于具有产卵表型的鹅。综上所述，数据表明miR-34c-5p通过靶向Bcl2调节颗粒细胞凋亡和育雏行为，这不仅有助于揭示miR-34c-5p的潜在机制鹅的潜在颗粒细胞凋亡，也为减少育雏发生率和提高产蛋量提供了有效策略。