Skip to main content

C-type natriuretic peptide inhibits porcine oocyte meiotic resumption

  • Yuki Hiradate (a1), Yumi Hoshino (a2), Kentaro Tanemura (a2) and Eimei Sato (a2)

C-type natriuretic peptide (CNP) is a recently identified meiotic inhibitor in mice. However, it has not been investigated in porcine oocytes to date. This study aimed to demonstrate the inhibitory effect of CNP against germinal vesicle breakdown (GVBD) in porcine oocyte meiotic resumption. Immunohistochemical analysis revealed intense natriuretic peptide receptor 2 (NPR2) immunoreactivity in the oocyte surrounded cumulus cells in the follicles. Furthermore, reverse transcription polymerase chain reaction (RT-PCR) analysis showed the expression of npr2 mRNA only in cumulus cells but not in oocytes, suggesting that cumulus cells are the targets of CNP. When cumulus–oocyte complexes (COCs) or denuded oocytes (DOs) were cultured with various concentrations of CNP (10, 50, 100, 500, and 1,000 nM), inhibitory effect was observed in the COC group, but not in the DO group, confirming that CNP indirectly inhibits GVBD via cumulus cells. This evidence is the first indication that the CNP-NPR2 pathway is involved in meiotic arrest in porcine oocytes. Furthermore, we investigated the effect of oocyte-derived paracrine factor (ODPF) on npr2 mRNA expression level in cumulus cells by evaluating changes in mRNA expression in oocytectomised COCs (OXCs) by real-time PCR. A significant decrease in npr2 mRNA expression level was observed in OXCs, whereas mRNA expression level was restored in OXCs with DOs, indicating that ODPF participates in the regulation of npr2 expression in porcine cumulus cells.

Corresponding author
All correspondence to Yuki Hiradate. Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981–8555, Japan. Tel:/Fax: +81 22 717 8687. E-mail:
Hide All
Ballermann, B.J. & Brenner, B.J (1987). Atrial natriuretic peptide and kidney. Am. J. Kidney Dis. 10, 712.
Downs, S.M., Coleman, D.L., Ward-Bailey, P.F. & Eppig, J.J. (1985). Hypoxanthine is the principal inhibitor of murine oocyte maturation in a low molecular weight fraction of porcine follicular fluid. Proc. Natl. Acad. Sci. USA 82, 454–8.
Eppig, J.J. & Downs, S.M. (1987). The effect of hypoxanthine on mouse oocyte growth and development in vitro: maintenance of meiotic arrest and gonadotropin-induced oocyte maturation. Dev. Biol. 119, 313–21.
Fan, H.Y., Huo, L.J., Chen, D.Y., Schatten, H. & Sun, Q.Y. (2004). Protein kinase C and mitogen-activated protein kinase cascade in mouse cumulus cells: cross talk and effect on meiotic resumption of oocyte. Biol. Reprod. 70, 1178–87.
Gotze, M., Kauffold, P., Schuffenhauer, A., Torner, H. & Spitschak, M. (1990). [The inhibition of meiosis of bovine oocytes using biologic of synthetic inhibitors.] Arch. Exp. Veterinarmed. 44, 1927.
Jankowski, M., Reis, A.M., Mukkadam-Daher, S., Dam, T.V., Farookhi, R. & Gutkowska, J. (1997). C-type natriuretic peptide and the guanylyl cyclase receptors in the rat ovary modulated by the estrous cycle. Biol. Reprod. 56, 5966.
Kadam, A.L. & Koide, S.S. (1990). Identification of hypoxanthine in bovine follicular fluid. J. Pharm. Sci. 79, 1077–82.
Kimura, N., Konno, Y., Miyoshi, K., Matsumoto, H. & Sato, E. (2002). Expression of hyaluronan synthases and CD44 messenger RNAs in porcine cumulus–oocyte complexes during in vitro maturation. Biol. Reprod. 66, 707–17.
Kiyosu, C., Tsuji, T., Yamada, K., Kajita, S. & Kunieda, T. (2012). NPPC/NPR2 signaling is essential for oocyte meiotic arrest and cumulus oophorus formation during follicular development in the mouse ovary. Reproduction 144, 187–93.
Norris, R.P., Ratzan, W.J., Freudzon, M., Mehlmann, L.M., Krall, J., Movsesian, M.A., Wang, H., Ke, H., Nikolaev, V.O. & Jaffe, L.A. (2009). Cyclic GMP from the surrounding somatic cells regulates cyclic AMP and meiosis in the mouse oocyte. Development 136, 1869–78.
Petr, J., Rajmon, R., Chmelíková, E., Tománek, M., Lánská, V., Pribánová, M. & Jílek, F. (2006). Nitric-oxide-dependent activation of pig oocytes: the role of the cGMP-signalling pathway. Zygote 14, 916.
Petters, R.M. & Wells, K.D. (1993). Culture of pig embryos. J. Reprod. Fertil. Suppl. 48, 6173.
Pincus, G. & Enzmann, E.V. (1935). The comparative behavior of mammalian eggs in vivo and in vitro: I. The activation of ovarian eggs. J. Exp. Med. 62, 665–75.
Potter, L.R., Abbey-Hosch, S. & Dickey, D.M (2006). Natriuretic peptides, their receptors, and cyclic guanosine monophosphate-dependent signaling functions. Endocr. Rev. 27, 4772.
Rahmutula, D. & Gardner, D.G. (2005). C-type natriuretic peptide down-regulates expression of its cognate receptor in rat aortic smooth muscle cells. Endocrinology 146, 4968–74.
Roh, S.G., Song, S.H., Cho, K.C., Katoh, K., Wittamer, V., Parmentier, M. & Sasaki, S. (2007). Chemerin—a new adipokine that modulates adipogenesis via its own receptor. Biochem. Biophys. Res. Commun. 362, 1013–8.
Sato, Y., Cheng, Y., Kawamura, K., Takae, S. & Hsueh, A.J. (2012). C-type natriuretic peptide stimulates ovarian follicle development. Mol. Endocrinol. 26, 1158–66.
Schmidt, H., Stonkute, A., Jüttner, R., Schäffer, S., Buttgereit, J., Feil, R., Hofmann, F. & Rathjen, F.G. (2007 ). The receptor guanylyl cyclase Npr2 is essential for sensory axon bifurcation within the spinal cord. J. Cell. Biol. 179, 331–40.
Schulz, S. (2005). C-type natriuretic peptide and guanylyl cyclase B receptor. Peptides 26, 1024–34.
Törnell, J., Carlsson, B. & Billing, H. (1990). Atrial natriuretic peptide inhibits spontaneous rat oocyte maturation. Endocrinology 126, 1504–8.
Warikoo, P.K. & Bavister, B.D. (1989). Hypoxanthine and cyclic adenosine 5′-monophosphate maintain meiotic arrest of rhesus monkey oocytes in vitro. Fertil. Steril. 51, 886–9.
Webb, R.J., Marshall, F., Swann, K. & Carroll, J. (2002). Follicle-stimulating hormone induces a gap junction-dependent dynamic change in [cAMP] and protein kinase A in mammalian oocytes. Dev. Biol. 246, 441–54.
Woodard, G.E. & Rosado, J.A. (2008). Natriuretic peptides in vascular physiology and pathology. Int. Rev. Cell. Mol. Biol. 268, 5993.
Zhang, M., Su, Y.Q., Sugiura, K., Xia, G. & Eppig, J.J. (2010). Granulosa cell ligand NPPC and its receptor NPR2 maintain meiotic arrest in mouse oocytes. Science 330, 366–9.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

  • ISSN: 0967-1994
  • EISSN: 1469-8730
  • URL: /core/journals/zygote
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed