Suppression of Notch Signaling Stimulates Progesterone Synthesis by Enhancing the Expression of NR5A2 and NR2F2 in Porcine Granulosa Cells

• Published in: Genes Vol. 11; no. 2; p. 120
• Main Authors: Guo, Rihong, Chen, Fang, Shi, Zhendan
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 22.01.2020; MDPI AG
• Subjects: Animals; COUP Transcription Factor II - genetics; COUP Transcription Factor II - metabolism; Dipeptides - pharmacology; Female; granulosa cells; Granulosa Cells - metabolism; Lipogenesis - physiology; notch signaling; nr2f2; nr5a2; porcine; Primary Cell Culture; progesterone; Progesterone - biosynthesis; Progesterone - metabolism; Receptor, Notch3 - drug effects; Receptor, Notch3 - metabolism; Receptors, Cytoplasmic and Nuclear - genetics; Receptors, Cytoplasmic and Nuclear - metabolism; Receptors, Notch - drug effects; Receptors, Notch - metabolism; Signal Transduction; Swine; porcine; NR5A2; NR2F2; granulosa cells; Notch signaling; progesterone
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes11020120

The conserved Notch pathway is reported to be involved in progesterone synthesis and secretion; however, the exact effects remain controversial. To determine the role and potential mechanisms of the Notch signaling pathway in progesterone biosynthesis in porcine granulosa cells (pGCs), we first used a pharmacological γ-secretase inhibitor, -( -(3,5-difluorophenacetyl-l-alanyl))-S-phenylglycine t-butyl ester (DAPT), to block the Notch pathway in cultured pGCs and then evaluated the expression of genes in the progesterone biosynthesis pathway and key transcription factors (TFs) regulating steroidogenesis. We found that DAPT dose- and time-dependently increased progesterone secretion. The expression of steroidogenic proteins NPC1 and StAR and two TFs, NR5A2 and NR2F2, was significantly upregulated, while the expression of HSD3B was significantly downregulated. Furthermore, knockdown of both and with specific siRNAs blocked the upregulatory effects of DAPT on progesterone secretion and reversed the effects of DAPT on the expression of NPC1, StAR, and HSD3B. Moreover, knockdown of NR5A2 and NR2F2 stimulated the expression of Notch3. In conclusion, the inhibition of Notch signaling stimulated progesterone secretion by enhancing the expression of NPC1 and StAR, and the two TFs NR5A2 and NR2F2 acted as downstream TFs of Notch signaling in regulating progesterone synthesis.