Bravo R, Otero C, Allende C C, Allende J E
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1242-6. doi: 10.1073/pnas.75.3.1242.
Two inhibitors of cyclic AMP phosphodiesterase (3':5'-cyclic-AMP 5'-nucleotidohydrolase, EC 3.1.4.17), theophylline and papaverine, inhibit the maturation of Xenopus laevis oocytes induced by four different stimuli: human chorionic gonadotropin, progesterone, testosterone, and lanthanum ions. Addition of 1 mM cyclic AMP to the medium delays maturation by approximately 2 hr. Papaverine, theophylline, and cyclic AMP inhibit amino acid incorporation into oocyte proteins by 50% or more but do not inhibit amino acid uptake. The capacity of theophylline to block maturation and protein synthesis is reversed in a parallel fashion by addition of 1-5 mM calcium ion to the medium. Addition of papaverine, theophylline, and cycloheximide to oocytes at different times after hormonal treatment shows that the step sensitive to blockage by the three drugs is coincident and precedes germinal vesicle breakdown by about 1.5 hr. Theophylline and papaverine do not increase endogenous cyclic AMP levels in oocytes but do block the decrease of cyclic AMP levels observed 3 hr after progesterone treatment. Both drugs inhibit oocyte cyclic AMP phosphodiesterase measured in vivo and severely inhibit the stimulus of calcium uptake caused by progesterone and human chorionic gonadotropin. These results suggest that cyclic AMP, theophylline, and papaverine may block oocyte maturation by inhibiting protein synthesis, possibly via a cyclic AMP-dependent protein kinase as shown in reticulocytes [Datta, A., De Haro, C., Sierra, J. & Ochoa, S. (1977) Proc. Natl. Acad. Sci., USA 74, 1463-1467].
两种环磷酸腺苷磷酸二酯酶(3':5'-环磷酸腺苷5'-核苷酸水解酶,EC 3.1.4.17)抑制剂,即茶碱和罂粟碱,可抑制由四种不同刺激诱导的非洲爪蟾卵母细胞成熟,这四种刺激分别是人绒毛膜促性腺激素、孕酮、睾酮和镧离子。向培养基中添加1 mM环磷酸腺苷可使成熟延迟约2小时。罂粟碱、茶碱和环磷酸腺苷可使氨基酸掺入卵母细胞蛋白质的量减少50%或更多,但不抑制氨基酸摄取。向培养基中添加1 - 5 mM钙离子可平行逆转茶碱阻断成熟和蛋白质合成的能力。在激素处理后的不同时间向卵母细胞中添加罂粟碱、茶碱和环己酰亚胺,结果表明这三种药物敏感的阻断步骤是一致的,且在生发泡破裂前约1.5小时。茶碱和罂粟碱不会增加卵母细胞内源性环磷酸腺苷水平,但会阻断孕酮处理3小时后观察到的环磷酸腺苷水平的下降。这两种药物均可抑制体内测得的卵母细胞环磷酸腺苷磷酸二酯酶,并严重抑制孕酮和人绒毛膜促性腺激素引起的钙摄取刺激。这些结果表明,环磷酸腺苷、茶碱和罂粟碱可能通过抑制蛋白质合成来阻断卵母细胞成熟,可能是通过如网织红细胞中所示的依赖环磷酸腺苷的蛋白激酶[达塔,A.,德哈罗,C.,西拉,J. & 奥乔亚,S.(1977年)美国国家科学院院刊74,1463 - 1467]。