Atrial natriuretic peptide negatively regulates follicle-stimulating hormone-induced porcine oocyte maturation and cumulus expansion via cGMP-dependent protein kinase pathway.
This study examined the effect of atrial natriuretic peptide (ANP) on porcine cumulus-enclosed oocyte (CEO) maturation and cumulus expansion. ANP negatively regulated follicle-stimulating hormone (FSH)-stimulated germinal vesicle breakdown (GVBD; 90.1, 81.2 and 68.2% for FSH, FSH+10nM ANP and FSH+1 microM ANP, respectively), first polar body emission (PB1; 86.1, 75.3 and 53.3% for FSH, FSH+1 nM ANP and FSH+1 microM ANP, respectively) and cumulus expansion (CEI; 3.47, 3.16 and 2.43 for FSH, FSH+1 nM ANP and FSH+1 microM ANP, respectively) in a dose-dependent manner when CEOs were cultured in the maturation medium containing porcine follicular fluid (pFF). This negative effect showed a time-dependent manner after preincubation with 100 nM ANP for 5h (78.4% PB1), 10h (81.7% GVBD and 74.1% PB1), 20 h (78.5% GVBD and 68.9% PB1), and 44 h (75.3% GVBD and 60.5% PB1), respectively. ANP also significantly inhibited FSH-induced porcine oocyte GVBD (47.6% versus 83.8%) and PB1 emission (22.4% versus 45.2%) when CEOs were cultured in pFF-free maturation medium. cGMP analog 8-Br-cGMP (10 microM to 1mM) mimicked the effects of ANP on GVBD, PB1, and CEI. The negative effect of ANP was completely reversed by KT5823 (a specific inhibitor of cGMP-dependent protein kinase), while C-ANP-(4-23) (an analogue of ANP and specific binder for natriuretic peptide receptors-C) was ineffective in oocyte maturation. Neither ANP nor C-ANP-(4-23) had an effect on spontaneous porcine oocyte maturation and cumulus expansion. These results suggested that ANP negatively regulates FSH-activated porcine oocyte meiotic resumption, meiotic maturation and cumulus expansion. The function of ANP on porcine oocyte maturation is via the cGMP dependent protein kinase (PKG) pathway.
Zhang
Meijia
M
Department of Animal Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, PR China.
Tao
Yong
Y
Xia
Guoliang
G
Xie
Huirong
H
Hong
Haiyan
H
Wang
Fengchao
F
Lei
Lei
L
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000494:pharmacology*; D002227:Carbazoles / Q000494:pharmacology; D002478:Cells, Cultured; D003470:Culture Media; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development*; D006080:Ovarian Follicle / Q000187:drug effects / Q000502:physiology*; D010446:Peptide Fragments / Q000494:pharmacology; D013552:Swine
D002227:Carbazoles; D003470:Culture Media; D007211:Indoles; D010446:Peptide Fragments; C065367:atrial natriuretic factor (4-23)NH2, de-Gln(18)-de-Ser(19)-de-Gly(20,22)-de-Leu(21)-; C073601:KT 5823; C016276:8-bromocyclic GMP; D009320:Atrial Natriuretic Factor; D005640:Follicle Stimulating Hormone; D017869:Cyclic GMP-Dependent Protein Kinases; D006152:Cyclic GMP