Kotani E, Usuki S, Kubo T
Department of Obstetrics and Gynecology, University of Tsukuba, Ibaraki, Japan.
J Reprod Fertil. 1998 Jan;112(1):107-14. doi: 10.1530/jrf.0.1120107.
The purpose of this study was to detect the presence of mRNA encoding pituitary adenylate cyclase-activating polypeptide (PACAP) type I receptor in granulosa cells from rat ovary and to examine the effect of PACAP on progestin biosynthesis. mRNA was isolated from granulosa cells from the ovaries of immature rats treated with pregnant mares' serum gonadotrophin. The technique of reverse transcription and polymerase chain reaction with primers specific to PACAP type I receptor were used to demonstrate the expression of mRNA encoding PACAP type IA receptor in these cells. Granulosa cells were also cultured in the absence or presence of 100 ng LH ml-1 with various doses of PACAP-38 (10, 100 and 1000 ng ml-1). At the end of the incubation period, the incubation media were collected and concentrations of progesterone, 20 alpha-hydroxypregn-4-en-3-one (20 alpha-OH-P) and cAMP were measured. Increasing concentrations of PACAP-38 significantly stimulated the production of progestins (progesterone and 20 alpha-OH-P) and cAMP accumulation in a dose-dependent manner (P < 0.01; ANOVA). This effect was observed in media cultured for 24 and 48 h in both basal and LH-stimulated states. PACAP-38 did not significantly affect the ratio of progesterone: 20 alpha-OH-P produced by granulosa cells cultured for 24 h in the LH-stimulated state. However, at 1000 ng ml-1, PACAP-38 significantly decreased the ratio of progesterone to 20 alpha-OH-P production in granulosa cells cultured for 48 h (P < 0.01). These results suggest that granulosa cells from rat ovary express mRNA encoding PACAP type IA receptor and that PACAP may regulate granulosa cell differentiation and play an important role in the reproductive process.
本研究的目的是检测大鼠卵巢颗粒细胞中编码I型垂体腺苷酸环化酶激活多肽（PACAP）受体的mRNA的存在情况，并研究PACAP对孕激素生物合成的影响。从未成熟大鼠卵巢中提取颗粒细胞，这些大鼠用孕马血清促性腺激素处理过。采用逆转录和聚合酶链反应技术，使用针对I型PACAP受体的特异性引物，以证明这些细胞中编码IA型PACAP受体的mRNA的表达。颗粒细胞也在不存在或存在100 ng LH ml-1以及不同剂量的PACAP-38（10、100和1000 ng ml-1）的情况下进行培养。在孵育期结束时，收集孵育培养基并测量孕酮、20α-羟基孕-4-烯-3-酮（20α-OH-P）和cAMP的浓度。PACAP-38浓度的增加以剂量依赖的方式显著刺激了孕激素（孕酮和20α-OH-P）的产生以及cAMP的积累（P < 0.01；方差分析）。在基础状态和LH刺激状态下培养24小时和48小时的培养基中均观察到了这种效应。在LH刺激状态下培养24小时的颗粒细胞中，PACAP-38对孕酮与20α-OH-P的产生比例没有显著影响。然而，在1000 ng ml-1时，PACAP-38显著降低了在LH刺激状态下培养48小时的颗粒细胞中孕酮与20α-OH-P的产生比例（P < 0.01）。这些结果表明，大鼠卵巢颗粒细胞表达编码IA型PACAP受体的mRNA，并且PACAP可能调节颗粒细胞分化并在生殖过程中发挥重要作用。
