Promoter Identification and Transcriptional Regulation of the Goose <i>AMH</i> Gene

• Main Authors: Shuang Yang, Yan Deng, Da Chen, Shenqiang Hu, Yingying Zhang, Huilan Huang, Jiwei Hu, Liang Li, Hua He, Jiwen Wang
• Format: Article
• Language: English
• Published: MDPI AG 2019-10-01
• Series: Animals
• Subjects: goose; amh; granulosa cells; promoter analysis; transcriptional regulation
• Online Access: https://www.mdpi.com/2076-2615/9/10/816

Anti-M&#252;llerian hormone (AMH) is recognized as a reliable marker of ovarian reserve. However, the regulatory mechanism of goose <i>AMH</i> gene remains poorly understood. In the present study, both the full-length coding sequence (CDS) and promoter sequence of goose <i>AMH</i> have been cloned. Its CDS consisted of 2013 nucleotides encoding 670 amino acids and the amino acid sequence contained two structural domain: AMH-N and transforming growth factor beta (TGF-&#946;) domain. The obtained promoter sequence spanned from the &#8722;2386 bp to its transcription start site (ATG). Core promoter regions and regulatory elements were identified as well as transcription factors were predicted in its promoter sequence. The luciferase activity was the highest spanning from the &#8722;331 to &#8722;1 bp by constructing deletion promoter reporter vectors. In CHO cells, the luciferase activity significantly increased by co-expression of <i>AMH</i> and GATA binding protein 4 (<i>GATA-4</i>), while that significantly decreased by mutating the binding sites of <i>GATA-4</i> located in the &#8722;778 and &#8722;1477 bp. Results from quantitative real-time polymerase chain reaction (qPCR) indicated that levels of <i>AMH</i> mRNA in geese granulosa layers decreased gradually with the increasing follicular diameter. Taken together, it could be concluded that the transcriptional activity of <i>AMH</i> was activated by <i>GATA-4</i> to inhibit the development of small follicles in goose.