Regulatory mechanisms for natriuretic peptide signalling in sheep granulosa cells

Alhamdan, Rana; Maalouf, Walid E.; Campbell, Bruce K.; Hernandez-Medrano, Juan; Marsters, Peter

Abstract

Natriuretic peptides (NPs) have been reported to have critical roles in follicular development and oocyte maturation in rodents. This study aimed to extend our current understanding of NP-mediated signalling pathways and mechanisms of action in the follicles of a monovulatory species. Ovine granulosa cells (GCs) and theca cells (TCs) were cultured under conditions designed to allow gonadotrophin-stimulated cell differentiation. Gene expression analysis was performed by qualitative (q)PCR for NPs and NPRs (between 16 and 96 h of culture) and VEGF120 and VEGF164 (between 16 and 144 h of culture). A qualitative analysis of the production of NP/NPR family members and NP ligand/receptor associations was carried out utilising a highly sensitive immunological approach known as ‘proximity ligation assay’ (PLA). All NPRs were observed in GCs, while NPRA was absent in TCs. In GCs, gene expression of NPRA, NPRB and NPRC was apparent but only active BNP and CNP and not ANP, were detected. Also in GCs, ANP but not CNP was able to significantly (P [less than] 0.05) reduce oestradiol and increase (P [less than] 0.05) progesterone. Inhibition of VEGF164 by ANP and CNP (P [less than] 0.01) after 48 h of culture preceded up-regulation of VEGF120 by ANP (P [less than] 0.01) after 144 h, but not CNP. Taken together, these findings appear to demonstrate that NP responsiveness in the GC compartment of sheep follicles is multi-facilitated, utilising both autocrine and paracrine stimulation pathways.

Citation

Alhamdan, R., Maalouf, W. E., Campbell, B. K., Hernandez-Medrano, J., & Marsters, P. (2020). Regulatory mechanisms for natriuretic peptide signalling in sheep granulosa cells. Journal of Molecular Endocrinology, 64(4), 223-234. https://doi.org/10.1530/jme-20-0034

• Journal Article Type: Article
• Acceptance Date: Mar 6, 2020
• Online Publication Date: May 1, 2020
• Publication Date: May 1, 2020
• Deposit Date: Mar 30, 2020
• Publicly Available Date: May 2, 2021
• Journal: Journal of Molecular Endocrinology
• Print ISSN: 0952-5041
• Electronic ISSN: 1479-6813
• Publisher: BioScientifica
• Peer Reviewed: Peer Reviewed
• Volume: 64
• Issue: 4
• Pages: 223-234
• DOI: https://doi.org/10.1530/jme-20-0034
• Keywords: Molecular Biology; Endocrinology
• Public URL: https://nottingham-repository.worktribe.com/output/4218749
• Publisher URL: https://jme.bioscientifica.com/view/journals/jme/64/4/JME-20-0034.xml
• Additional Information: Disclaimer: this is not the definitive Version of Record of this article. This manuscript has been accepted for publication in Journal of Molecular Endocrinology , but the version presented here has not yet been copy-edited, formatted or proofed. Consequently, Bioscientifica accepts no responsibility for any errors or omissions it may contain. The definitive version is now freely available at ; https://doi.org/10.1530/JME-20-0034 May 2020.