The effects of mouse ovarian granulosa cell function and related gene expression by suppressing BMP/Smad signaling pathway

Li Zhang; Hejian Wang; Daolun Yu; Jie Chen; Chaofeng Xing; Jie Li; Jun Li; Yafei Cai

doi:10.1080/19768354.2018.1497706

The effects of mouse ovarian granulosa cell function and related gene expression by suppressing BMP/Smad signaling pathway

Anim Cells Syst (Seoul). 2018 Sep 19;22(5):317-323. doi: 10.1080/19768354.2018.1497706. eCollection 2018.

Authors

Li Zhang¹, Hejian Wang¹, Daolun Yu¹, Jie Chen¹, Chaofeng Xing¹, Jie Li¹, Jun Li¹, Yafei Cai²

Affiliations

¹ College of Life Science, Anhui provincial Key Lab of the Conservation and Exploitation of Biological Resources, Anhui Normal University, Wuhu, People's Republic of China.
² College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.

Abstract

BMP I type receptor inhibitor can selectively inhibit BMP/Smad signaling pathways, mainly by inhibiting the BMP I type receptor activity to prevent phosphorylation of Smad1, Smad5 and Smad9. The aim of the present study was to explore the effects of mouse ovarian granulosa cell function and related gene expression by suppressing BMP/Smad signaling pathway with LDN-193189(A type of BMP I type receptor inhibitor). In this study, we cultivate the original generation of mouse ovarian granular cells then collect cells and cell culture medium after treatment. Cellular localization and expression of Smad9 and P-smad9 proteins was studied by immunofluorescence (IF) in the ovarian granulosa cells of mouse; Related genes mRNA and proteins expression was checked by QRT-PCR and Western blot; Detected the concentration of related hormones by using ELISA kit; finally, the growth of the cells was analyzed by plotting cell growth curve with CCK-8 assay. The results indicate that, suppression of BMP/Smad signaling pathway can inhibit the expression of LHR and FSHR, inhibit cell proliferation and decrease E₂ secretion, the mechanism of action maybe reduce the expression of smad9, at the same time, we found that the feedback regulation of smad9 may affect the expression of FSHR and cell proliferation.

Keywords: BMP-4; BMP/Smad signal pathway; Granulosa cells; Smad9.

Grants and funding

This study was supported by the National Natural Science Foundation of China (NSFC) [grant number 31572265 and 81570094]; Jiangsu Agriculture Science and Technology Innovation Fund [grant number CX(18)3014].