Suzuki S, Endo Y, Fujiwara T, Tanaka S, Iizuka R
Fertil Steril. 1983 May;39(5):683-9.
The aim of this study was to determine whether steroidogenesis occurs in human immature oocytes aspirated from follicles during gynecologic laparotomy. delta 5-3 beta-Hydroxysteroid dehydrogenase (delta 5-3 beta-HSD) and 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) activities were detected by Dickmann and Dey's reaction medium consisting of 1.8 mg substrate (pregnenolone or 17 beta-estradiol [E2]), 4 mg nicotinamide-adenine dinucleotide, 2 mg nitro-blue tetrazolium, 10 ml 0.1 M phosphate buffer. The activity of adenylate cyclase was examined by ultrastructural-cytochemical study using 5'-adenylyl-imidodiphosphate (AMP-PNP) as a substrate in Vorbrodt's medium of 0.005 M AMP-PNP, 0.001 M MgCl2, 0.02 M SrCl2, 0.01 M NaF, 0.002 M theophylline, 0.01 M Tris-HCl. Furthermore, in indirect immunofluorescence study, the presence of endogenous progesterone and E2 and the activity of delta 5-3 beta-HSD were demonstrated. The results suggest that steroidogenesis, through the adenylate cyclase-cyclic adenosine 3':5'-monophosphate system, in human oocytes may play some important role in oocyte maturation, fertilization, and early embryonic development. The implication of steroid-producing activities of the human oocytes for cytoplasmic maturation is discussed.
本研究的目的是确定在妇科剖腹手术中从卵泡吸出的人未成熟卵母细胞中是否发生类固醇生成。通过由1.8毫克底物(孕烯醇酮或17β-雌二醇[E2])、4毫克烟酰胺腺嘌呤二核苷酸、2毫克硝基蓝四唑、10毫升0.1M磷酸盐缓冲液组成的迪克曼和戴伊反应介质检测δ5-3β-羟基类固醇脱氢酶(δ5-3β-HSD)和17β-羟基类固醇脱氢酶(17β-HSD)活性。在由0.005M AMP-PNP、0.001M MgCl2、0.02M SrCl2、0.01M NaF、0.002M茶碱、0.01M Tris-HCl组成的沃布罗特介质中,以5'-腺苷酰-亚氨二磷酸(AMP-PNP)为底物,通过超微结构细胞化学研究检测腺苷酸环化酶的活性。此外,在间接免疫荧光研究中,证实了内源性孕酮和E2的存在以及δ5-3β-HSD的活性。结果表明,通过腺苷酸环化酶-环磷酸腺苷系统的类固醇生成在人卵母细胞中可能在卵母细胞成熟、受精和早期胚胎发育中发挥一些重要作用。讨论了人卵母细胞产生类固醇活性对细胞质成熟的意义。