Petr J, Rajmon R, Chmelíková E, Tománek M, Lánská V, Pribánová M, Jílek F
Research Institute of Animal Production, Prátelství 815, Prague 10, Czech Republic.
Zygote. 2006 Feb;14(1):9-16. doi: 10.1017/S0967199406003546.
Pig oocytes matured in vitro were parthenogenetically activated (78%) after treatment with 2 mM nitric oxide-donor (+/-)-S-nitroso-N-acetylpenicillamine (SNAP) for 24 h. Inhibition of soluble guanylyl cyclase with the specific inhibitors 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) or 6-anilino-5,8-quinolinequinone (LY83583) suppressed the SNAP-induced activation in a dose-dependent manner (23% of activated oocytes after treatment with 400 microM ODQ; 12% of activated oocytes after treatment with 40 microM LY83583). 8-Bromo-cyclic guanosine monophosphate (8-Br-cGMP), a phosphodiesterase-resistant analogue of cGMP, enhances the effect of suboptimal doses (0.1 or 0.5 mM) of the NO donor SNAP. DT3, a specific inhibitor of cGMP-dependent protein kinase (PKG, PKG), is also able to inhibit the activation of pig oocytes after NO donor treatment. Involvement of the cGMP-dependent signalling pathway is specific for NO-induced oocyte activation, because both the guanylyl cyclase inhibitor ODQ and the PKG inhibitor DT3 are unable to inhibit activation in oocytes treated with the calcium ionophore A23187. These data indicate that the activation of pig oocytes with an NO donor is cGMP-dependent and that PKG plays an important role in this mode of oocyte activation.
用2 mM一氧化氮供体（±）-S-亚硝基-N-乙酰青霉胺（SNAP）处理24小时后，体外成熟的猪卵母细胞发生了孤雌激活（激活率78%）。用特异性抑制剂1H-[1,2,4]恶二唑并[4,3-a]喹喔啉-1-酮（ODQ）或6-苯胺基-5,8-喹啉醌（LY83583）抑制可溶性鸟苷酸环化酶，可剂量依赖性地抑制SNAP诱导的激活（用400 microM ODQ处理后，23%的卵母细胞被激活；用40 microM LY83583处理后，12%的卵母细胞被激活）。8-溴环鸟苷单磷酸（8-Br-cGMP）是一种抗磷酸二酯酶的cGMP类似物，可增强次优剂量（0.1或0.5 mM）的一氧化氮供体SNAP的作用。DT3是一种cGMP依赖性蛋白激酶（PKG）的特异性抑制剂，它也能够抑制一氧化氮供体处理后猪卵母细胞的激活。cGMP依赖性信号通路参与一氧化氮诱导的卵母细胞激活具有特异性，因为鸟苷酸环化酶抑制剂ODQ和PKG抑制剂DT3均不能抑制用钙离子载体A23187处理的卵母细胞的激活。这些数据表明，用一氧化氮供体激活猪卵母细胞是cGMP依赖性的，并且PKG在这种卵母细胞激活模式中起重要作用。
