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C/EBP? regulates Vegf gene expression in granulosa cells undergoing luteinization during ovulation in female rats.

ABSTRACT: The ovulatory LH-surge increases Vegf gene expression in granulosa cells (GCs) undergoing luteinization during ovulation. To understand the factors involved in this increase, we examined the roles of two transcription factors and epigenetic mechanisms in rat GCs. GCs were obtained from rats treated with eCG before, 4?h, 8?h, 12?h and 24?h after hCG injection. Vegf mRNA levels gradually increased after hCG injection and reached a peak at 12?h. To investigate the mechanism by which Vegf is up-regulated after hCG injection, we focused on C/EBP? and HIF1?. Their protein expression levels were increased at 12?h. The binding activity of C/EBP? to the Vegf promoter region increased after hCG injection whereas that of HIF1? did not at this time point. The C/EBP? binding site had transcriptional activities whereas the HIF1? binding sites did not have transcriptional activities under cAMP stimulation. The levels of H3K9me3 and H3K27me3, which are transcriptional repression markers, decreased in the C/EBP? binding region after hCG injection. The chromatin structure of this region becomes looser after hCG injection. These results show that C/EBP? regulates Vegf gene expression with changes in histone modifications and chromatin structure of the promoter region in GCs undergoing luteinization during ovulation.

SUBMITTER: Shinagawa M

PROVIDER: S-EPMC6345775 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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C/EBPβ regulates Vegf gene expression in granulosa cells undergoing luteinization during ovulation in female rats.

Shinagawa Masahiro M Tamura Isao I Maekawa Ryo R Sato Shun S Shirafuta Yuichiro Y Mihara Yumiko Y Okada-Matsumoto Maki M Taketani Toshiaki T Asada Hiromi H Tamura Hiroshi H Sugino Norihiro N

Scientific reports 20190124 1

The ovulatory LH-surge increases Vegf gene expression in granulosa cells (GCs) undergoing luteinization during ovulation. To understand the factors involved in this increase, we examined the roles of two transcription factors and epigenetic mechanisms in rat GCs. GCs were obtained from rats treated with eCG before, 4 h, 8 h, 12 h and 24 h after hCG injection. Vegf mRNA levels gradually increased after hCG injection and reached a peak at 12 h. To investigate the mechanism by which Vegf is up-regu ...[more]

PMID: 30679486

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Project description:It has been known that EGF-like factor secreted from LH-stimulated granuloma cells acts on granulosa cells and cumulus cells to induce ovulation process. Granulosa cells are changed the morphology with differentiating cell functions to produce progesterone. Cumulus cells are detached to make a space between the cells to accumulate hyaluronan rich matrix. LH also changes extracellular matrix (ECM) components including fibronectin in the follicular walls and granulosa cell layers. EGF like factor and fibronectin synergistically play important roles in numerous cell functions, especially cancer cell migration, estimating that fibronectin would impact on granulosa cells and cumulus cells. To clear this hypothesis, the localizations of fibronectin and its receptor integrin were observed by immunofluorescence technique. The functions were monitored by the detection of downstream signaling pathway, focal adhesion kinase (FAK). The pharmacological approach in both in vivo and in vitro were used for analyzing the physiological roles of FAK during ovulation process. The immunofluorescence staining revealed that fibronectin and integrin were observed in granulosa cells, cumulus cells and the space between cumulus cells and oocyte at 4 and 8 h after hCG injection. Concomitantly with the changes of fibronectin-integrin localization, FAK was phosphorylated in periovulatory follicles. The injection of FAK inhibitor suppressed not only ovulation but also luteinization of granulosa cells and cumulus expansion. In cultured-granulosa cells, fibronectin-integrin synergistically activated FAK with amphiregulin (AREG). Such cooperative stimulations induced a morphological change in granulosa cells, which resulted in the maximum level of progesterone production via the induction of Hsd3b. When cumulus-oocyte complexes (COCs) were cultured with AREG in the presence of serum, the maximum level of cumulus expansion was observed. The AREG-induced cumulus expansion was also suppressed by FAK inhibitor. Thus, it is concluded that fibronectin and AREG synergistically activate FAK not only in granulosa cells and cumulus cells to induce successful ovulation process.

Dataset Information

C/EBP? regulates Vegf gene expression in granulosa cells undergoing luteinization during ovulation in female rats.

Publications

C/EBPβ regulates Vegf gene expression in granulosa cells undergoing luteinization during ovulation in female rats.

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