Abstract
ABSTRACTBMP 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.
Highlights
The steroid hormones secreted by ovarian granule cells (GCs) include progesterone, estradiol, luteinizing hormone, follicle-stimulating hormone, and aromatase cytochrome P450
First, extracellular signaling molecules Bone morphogenetic proteins (BMPs)-4 is combined with BMPR-II receptor, BMPR-I receptor activated by phosphorylated BMPR-II and forms a receptor complex with BMPR-II, activate I receptor protein kinase to make the intracellular Smads protein(such as smad1/5/9) phosphorylated, the activated r-smads combined with the common type of co-smad4 to form the Smad protein complex into the nucleus to regulate the transcription of the target gene (Drummond 2005)
BMP-4 is one of the bone morphogenetic proteins, it plays an important role in the process of embryonic development, and BMP-4 has important influence on female reproductive performance (Shimasaki et al 1999; Nilsson & Skinner 2003)
Summary
The steroid hormones secreted by ovarian granule cells (GCs) include progesterone, estradiol, luteinizing hormone, follicle-stimulating hormone, and aromatase cytochrome P450. Studies have shown that BMP4 proteins are expressed in ovarian GCs, theca cells and oocytes, and the expression level in all levels of follicle is different, change along with the development of follicles (Nilsson & Skinner 2003; Drummond 2005). It plays an important role in the maturation of oocyte and the growth and development of ovaries (Shimasaki et al 2004; Shimizu et al 2004; Juengel & McNatty 2005; Gilchrist et al 2008)
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