AUTHOR=Liu Yufang , Chen Yulin , Zhou Zuyang , He Xiaoyun , Tao Lin , Jiang Yanting , Lan Rong , Hong Qionghua , Chu Mingxing 

TITLE=chi-miR-324-3p Regulates Goat Granulosa Cell Proliferation by Targeting DENND1A

JOURNAL=Frontiers in Veterinary Science

VOLUME=Volume 8 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2021.732440

DOI=10.3389/fvets.2021.732440

ISSN=2297-1769

ABSTRACT=Granulosa cell (GC) proliferation provides essential conditions for ovulation in ani-mals. A previous study showed that DENND1A plays a significant role in polycystic ovary syndrome. However, the modulation of DENND1A in GCs remains unclear. Our previous integrated analysis of miRNA-mRNA revealed that the 3’ ’-untranslated region of DENND1A could be a target of chi-miR-324-3p. In this study, we used quantitative- reverse transcription- polymerase chain reaction (RT-qPCR) to investi-gate DENND1A expression in ovarian tissues of high- and low- yielding goats. Fur-ther, dual-fluorescent reporter vector experiments, Cell Counting Kit-8 (CCK-8) as-say, and RT-qPCR were used to elucidate the regulatory pathway of chi-miR-324-3p-DENND1A in GCs. The results revealed an opposite tendency between the expres-sions of chi-miR-324-3p and DENND1A in the ovaries of high- and low-yielding goats. The CCK-8 assay indicated that chi-miR-324-3p overexpression significantly suppressed GC proliferation, whereas chi-miR-324-3p inhibition promoted GC pro-liferation. In addition, the expressions of GC proliferation markers LHR, Cylin D2, and CDK4 showed the same tendency. The dual-fluorescent reporter assay revealed that chi-miR-324-3p directly targeted DENND1A, and the RT-qPCR results revealed that DENND1A expression was inhibited by chi-miR-324-3p. In summary, chi-miR-324-3p inhibited the proliferation of GCs by targeting DENND1A.