WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
C-type natriuretic peptide stimulates resumption of meiosis via a cGMP-dependant mechanism in porcine oocytes
Ryan D Rose 1 , Satoshi Sugimura 2 , Lesley J Ritter 1 , Hannah M Brown 1 , Jeremy G Thompson 1 & Robert B Gilchrist 3
1Robinson Research Institute, The University of Adelaide, Adelaide, South Australia, Australia; 2Tokyo University of Agriculture and Technology, Fuchu, Japan; 3University of New South Wales, Randwick, New South Wales, Australia.
Research conducted in recent years has led to great advances in our understanding of the participation of cGMP in meiosis. It is clear that increased intra-oocyte concentrations of cGMP inhibit meiosis in mouse models. Like cAMP, cGMP may also have a meiotic stimulatory function, possibly via cGMP/PKG. Abattoir derived gilt porcine ovaries were collected, antral follicles aspirated and oocytes collected and cultured in TCM-199+3 mg/ml BSA. Natriuretic peptide receptor 2 (NPR2) mRNA expression was two-fold higher in cumulus cells compared to granulosa cells. After 24 h of culture CNP increased the proportion of oocytes that resumed meiosis compared to control (main effect). Both 100 nM CNP and 1 μM 8pCPTcGMP (cGMP analogue) significantly reduced oocyte–cumulus gap junction communication (GJC), as measured by dye transfer, from 12 h by 1.7- and 1.9-fold respectively, compared to control (P<0.05). CNP or 8pCPTcGMP significantly increased ERK1/2 phosphorylation (western blot), 2.6- and 3.1-fold respectively, above control (P<0.05). This increase was eliminated by 10 μM AG1478 a selective EGF receptor inhibitor. Neither CNP nor 8pCPTcGMP had any effect on phosphorylation of cAMP-response binding protein (CREB) nor did they effect mRNA expression of EGF-like peptides at 2 h culture, compared to control. These results demonstrate that CNP stimulates the phosphorylation of ERK1/2, possibly via a cGMP-dependant mechanism requiring EGFR signalling in porcine COCs. This subsequently leads to a decrease in oocyte–cumulus GJC and the resumption of meiosis.
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ISSN 2052-1472 (online)
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