Botigelli Ramon Cesar, Schwarz Katia Lancellotti, Zaffalon Fabiane Gilli, Del Collado Maite, Castro Fernanda Cavallari, Fernandes Hugo, Leal Claudia Lima Verde
Departamento de Medicina Veterinária,Faculdade de Zootecnia e Engenharia de Alimentos,Universidade de São Paulo,Av. Duque de Caxias Norte,225,Pirassununga-SP,CEP 13635-900,Brasil.
Zygote. 2017 Jun;25(3):321-330. doi: 10.1017/S096719941700017X. Epub 2017 Jun 27.
This study aimed to examine the effects of nitric oxide (NO) and different phosphodiesterase (PDE) families on meiosis resumption, nucleotides levels and embryo production. Experiment I, COCs were matured in vitro with the NO donor S-nitroso-N-acetylpenicillamine (SNAP) associated or not with the soluble guanylate cyclase (sGC) inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), meiotic resumption and nucleotides levels were assessed. SNAP delayed germinal vesicle breakdown (GVBD) (53.4 ± 1.2 versus 78.4 ± 2.4% for controls, P 0.05). Cyclic GMP levels were higher in SNAP (3.94 ± 0.18, P 0.05). Embryo development did not differ from the control for SNAP and cilostamide groups (38.7 ± 5.8, 37.9 ± 6.2 and 40.5 ± 5.8%, P > 0.05), but SNAP + cilostamide decreased embryo production (25.7 ± 6.9%, P < 0.05). In conclusion, SNAP was confirmed to delay meiosis resumption by the NO/sGC/cGMP pathway, by increasing cGMP, but not cAMP. Inhibiting different PDEs to further increase nucleotides in association with SNAP did not show any additive effects on meiosis resumption, indicating that other pathways are involved. Moreover, SNAP + cilostamide affected the meiosis progression and decreased embryo development.
本研究旨在探讨一氧化氮(NO)和不同磷酸二酯酶(PDE)家族对减数分裂恢复、核苷酸水平及胚胎产生的影响。实验I,将卵丘-卵母细胞复合体(COCs)与NO供体S-亚硝基-N-乙酰青霉胺(SNAP)一起或不与可溶性鸟苷酸环化酶(sGC)抑制剂1H-[1,2,4]恶二唑并[4,3-a]喹喔啉-1-酮(ODQ)一起进行体外成熟培养,评估减数分裂恢复情况和核苷酸水平。SNAP延迟了生发泡破裂(GVBD)(对照组为78.4±2.4%,SNAP组为53.4±1.2%,P<0.05)。SNAP组的环磷酸鸟苷(cGMP)水平较高(3.94±0.18,P<0.05)。SNAP组和西洛他唑组的胚胎发育与对照组无差异(分别为38.7±5.8%、37.9±6.2%和40.5±5.8%,P>0.05),但SNAP+西洛他唑降低了胚胎产量(25.7±6.9%,P<0.05)。总之,证实SNAP通过NO/sGC/cGMP途径延迟减数分裂恢复,通过增加cGMP而非环磷酸腺苷(cAMP)来实现。与SNAP联合抑制不同的PDE以进一步增加核苷酸,对减数分裂恢复未显示出任何相加作用,表明涉及其他途径。此外,SNAP+西洛他唑影响减数分裂进程并降低胚胎发育。