Chen Rong, Chen Chen, Qin Yifei, Liu Jie, Lei Mingming, Zhu Huanxi, Shi Zhendan
Key Laboratory of Crop and Animal Integrated Farming, Ministry of Agriculture and Rural Affairs, Nanjing, PR China; Jiangsu Province Engineering Research Center of Precision Animal Breeding, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing, PR China.
Key Laboratory of Crop and Animal Integrated Farming, Ministry of Agriculture and Rural Affairs, Nanjing, PR China; Jiangsu Province Engineering Research Center of Precision Animal Breeding, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing, PR China.
Poult Sci. 2025 Feb;104(2):104752. doi: 10.1016/j.psj.2024.104752. Epub 2024 Dec 31.
Anti-Müllerian hormone (AMH) plays an important role in avian ovarian follicle development. The high mRNA expression of AMH in avian ovarian prehierarchical follicles helps prevent premature granulosa cell differentiation. Vitamin D3 was reported to downregulate AMH mRNA expression in granulosa cells of prehierarchical follicles in hens; however, the underlying molecular mechanism remains unknown. In this study, AMH mRNA expression was investigated in granulosa cells of prehierarchical follicles in geese under vitamin D3 induction. A potential vitamin D response element (VDRE) present in the goose AMH promoter was identified using luciferase activity, electrophoretic mobility shift assay (EMSA), and chromatin immunoprecipitation (ChIP) assays. The results showed that AMH mRNA expressions were downregulated by vitamin D3 in granulosa cells of prehierarchical follicles in geese. A classical VDRE-like sequence was identified in the goose AMH promoter by in silico analyses. Luciferase activity assays revealed that the putative VDRE is a negative regulatory element. Vitamin D receptor (VDR) and retinoic X receptor (RXR) are necessary to decrease the AMH promoter activity induced by vitamin D3. The EMSA and ChIP assays demonstrated that the VDR/RXR complex directly binds to the putative VDRE. These results suggest that vitamin D3 downregulates AMH mRNA expression via a functional VDRE that binds the VDR/RXR heterodimer in goose ovarian prehierarchical follicles. These results provide new insights into the regulatory mechanisms of vitamin D3 in avian ovarian follicular development.
抗苗勒管激素（AMH）在禽类卵巢卵泡发育中起重要作用。AMH在禽类卵巢前等级卵泡中的高mRNA表达有助于防止颗粒细胞过早分化。据报道，维生素D3可下调母鸡前等级卵泡颗粒细胞中AMH的mRNA表达；然而，其潜在的分子机制仍不清楚。在本研究中，研究了维生素D3诱导下鹅前等级卵泡颗粒细胞中AMH的mRNA表达。利用荧光素酶活性、电泳迁移率变动分析（EMSA）和染色质免疫沉淀（ChIP）分析，鉴定了鹅AMH启动子中存在的潜在维生素D反应元件（VDRE）。结果表明，维生素D3可下调鹅前等级卵泡颗粒细胞中AMH的mRNA表达。通过计算机分析在鹅AMH启动子中鉴定出一个经典的VDRE样序列。荧光素酶活性分析表明，推定的VDRE是一个负调控元件。维生素D受体（VDR）和视黄酸X受体（RXR）是降低维生素D3诱导的AMH启动子活性所必需的。EMSA和ChIP分析表明，VDR/RXR复合物直接与推定的VDRE结合。这些结果表明，维生素D3通过一个功能性VDRE下调AMH的mRNA表达，该VDRE在鹅卵巢前等级卵泡中与VDR/RXR异二聚体结合。这些结果为维生素D3在禽类卵巢卵泡发育中的调控机制提供了新的见解。
