ABSTRACT: Profitable animal production require high fertility animals that exhibit healthy ovarian follicles, which contributes to estradiol synthesis. Follicle maturation relies on coordination between pituitary hormones and intraovarian signaling pathways. Recently Wnt/beta-catenin signaling has been established as necessary modulators of folliculogenesis in response to FSH. In granulosa cells (GC), beta-catenin is a central key in regulation of aromatase (Cyp19a1) expression. In addition, protein kinase A (PKA) but not protein kinase B (AKT) phosphorylation of beta-catenin results in regulation of FSH target genes. A study conducted by our lab shown that WNT3A down regulate steroidogenesis, and recently another study confirmed that AKT is required in steroidogenesis.
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The objective of this study was to investigate if Glycogen Synthase Kinase-3 β (GSK3β) contributes for of cAMP and WNT3a phosphorylation on activation and subsequent gene regulation involved in steroidogenesis in GC. To evaluate the interaction between WNT and cAMP/PKA pathways, GC cultures were treated with or without PI3K inhibitor LY294002 (LY) (30 μM, 30 min), and with or without Forskolin (FSK) (10 μM, 1.5 h) and followed by WNT3a (50 ng/mL, 30 min). Cells were collected and total protein was isolated in M-PER and quantified by BCA kit. Western blot were used to detect total and phosphorylated
The objective of this study was to investigate if Glycogen Synthase Kinase-3 β (GSK3β) contributes for of cAMP and WNT3a phosphorylation on activation and subsequent gene regulation involved in steroidogenesis in GC. To evaluate the interaction between WNT and cAMP/PKA pathways, GC cultures were treated with or without PI3K inhibitor LY294002 (LY) (30 μM, 30 min), and with or without Forskolin (FSK) (10 μM, 1.5 h) and followed by WNT3a (50 ng/mL, 30 min). Cells were collected and total protein was isolated in M-PER and quantified by BCA kit. Western blot were used to detect total and phosphorylated