Effect of nitric oxide on the cyclic guanosine monophosphate (cGMP) pathway during meiosis resumption in bovine oocytes

Autor(es)

Lancellotti Schwarz, Katia R. ^[1] ; Lisboa Pires, Pedro R. ^[1] ; Mesquita, Ligia Garcia ^[1] ; Chiaratti, Marcos R. ^[2] ; Verde Leal, Claudia Lima ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Vet Med, Fac Zootecnia & Engn Alimentos, Pirassununga, SP - Brazil

^[2] Univ Fed Sao Carlos, Dept Genet & Evolucao, BR-13560 Sao Carlos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Theriogenology; v. 81, n. 4, p. 556-564, MAR 1 2014.
• Citações Web of Science: 13

Resumo

Nitric oxide (NO) is a chemical messenger involved in the control of oocyte maturation. It stimulates guanylate cyclase to produce cyclic guanosine monophosphate (cGMP), which in turn activates cGMP-dependent protein kinase (PKG) and some phosphodiesterases that may interfere with cAMP levels, a nucleotide also involved in meiosis resumption. The aim of this study was to determine the role played by NO on the cGMP/cAMP pathway during meiosis resumption in bovine oocytes. The effects of increasing NO generated by S-nitroso-N-acetylpenicillamine (SNAP; 10(-7)-10(-3) mol/L) and of other drugs that may affect the NO/cGMP pathway (proptoporfirin IX and 8-Br-cGMP) on meiosis resumption were investigated in bovine cumulus-oocyte complexes (COCs) matured for 9 hours in a semidefined medium (TCM199 + 3 mg/mL BSA). The COCs matured with 10(-7) mol/L SNAP associated or not with 100 mu mol/L oxadiazole-one quinoxaline, a guanylate cyclase inhibitor, also had their cGMP and CAMP levels measured during the first hours of maturation (1, 3, and 6 hours). Quantitative polymerase chain reaction was performed by real-time polymerase chain reaction to determine the effects of NO on expression of genes encoding for enzymes of the NO/guanylate cyclase/cGMP and CAMP pathways during the first 9 hours of oocyte maturation. Increasing NO levels using 10(-7) mol/L SNAP resulted in lower rate of germinal vesicle breakdown (36% germinal vesicle breakdown; P < 0.05) at 9 hours IVM, whereas control group and the treatments with 10(-9) and 10(-8) mol/L SNAP showed about 70% germinal vesicle breakdown (P > 0.05). A temporary increase in cGMP levels was also observed with the same treatment (4.51 pmol/COC) at 1 hour IVM, which was superior to the control group (2.97 pmol/COC; P < 0.05) and was reversed by inhibiting guanylate cyclase activity with 100 mu mol/L oxadiazole-one quinoxaline. Neither cAMP levels nor gene expression were affected by NO. These results suggest that NO acts via guanylate cyclase/cGMP and that even a temporary increase in cGMP levels leads to a delay in meiosis resumption, even when cAMP levels have declined. Nitric oxide does not act on oocyte maturation by affecting cAMP levels or the expression of genes related to the NO/guanylate cyclase/cGMP and cAMP pathways. Also, to our knowledge this is the first report to detect PKG1, PKG2, phosphodiesterase-5A, ADCY3, ADCY6, and ADCY9 transcripts in bovine oocytes. (C) 2014 Elsevier Inc. All rights reserved. (AU)

• Processo FAPESP: 08/09321-6 - O hormônio folículo estimulante e a via da óxido nítrico sintase na maturação de oócitos bovinos
• Beneficiário: Cláudia Lima Verde Leal
• Modalidade de apoio: Auxílio à Pesquisa - Regular