Gräs S, Hannibal J, Georg B, Fahrenkrug J
Department of Clinical Biochemistry, University of Copenhagen, Bispebjerg Hospital, Denmark.
Endocrinology. 1996 Nov;137(11):4779-85. doi: 10.1210/endo.137.11.8895347.
Pituitary adenylate cyclase activating polypeptide (PACAP), a potent stimulator of cAMP formation, has a widespread distribution in neuronal elements of the central nervous system and a number of peripheral organs. Recently, PACAP has also been shown to be stage specifically expressed in the spermatogenic cells from the rat testis. This prompted us to examine the cell-specific expression of PACAP during the estrous cycle in the rat ovary by in situ hybridization with a digoxiginin UTP-labeled RNA probe and by immunohistochemistry using a specific monoclonal antibody. Ovaries from PMS gonadotropin (PMSG)/human CG (hCG) treated immature animals were examined 0, 12, 24, 48, 54, 57, 58.5, 60, 61.5, 63, 66, 72, and 96 h after PMSG treatment. Ovaries from adult cyclic rats were examined at 1000 h on each day of the estrous cycle and at 0400 h on estrus morning. PACAP positive cells were observed in the preovulatory and ovulatory period, i.e. 6-18 h following hCG injection in the PMSG induced cycle and at 0400 h and 1000 h on estrus morning in adult cyclic animals. Both PACAP messenger RNA (mRNA) and PACAP immunoreactivity were observed in the majority of granulosa and cumulus cells from large preovulatory follicles, in the majority of the cells comprising the interstitial glandular tissue and in solitary theca cells of growing and mature follicles. In PMSG/hCG treated animals, positive signals were also observed in solitary theca and interstitial cells 12 h after PMSG injection. PACAP immunoreactive nerve fibers were observed in all ovaries examined, located in the interstitial glandular tissue or in the loose connective tissue. Northern blotting of ovarian tissue confirmed the transient preovulatory and ovulatory expression of PACAP. Three transcripts were observed. The most predominant band had a size of approximately 1.2 kilobase pairs, corresponding to the transcript observed in the testis. Two bands of 2.4 and 3.0 kilobase pairs, corresponding to the transcripts from neural tissue, were also observed. The complex distribution of PACAP positive cells and nerve fibers suggests that the peptide could be a local regulator of a number of the events that take place in the periovulatory period as well as during early folliculogenesis in the ovary.
垂体腺苷酸环化酶激活多肽（PACAP）是一种有效的环磷酸腺苷（cAMP）形成刺激剂，广泛分布于中枢神经系统的神经元以及许多外周器官中。最近，PACAP还被证明在大鼠睾丸的生精细胞中呈阶段特异性表达。这促使我们通过用地高辛标记的UTP RNA探针进行原位杂交以及使用特异性单克隆抗体进行免疫组织化学，来检测大鼠卵巢发情周期中PACAP的细胞特异性表达。对经孕马血清促性腺激素（PMSG）/人绒毛膜促性腺激素（hCG）处理的未成熟动物的卵巢，在PMSG处理后的0、12、24、48、54、57、58.5、60、61.5、63、66、72和96小时进行检查。对成年周期大鼠的卵巢，在发情周期的每一天的1000时以及发情日早晨的0400时进行检查。在排卵前和排卵期观察到PACAP阳性细胞，即在PMSG诱导周期中注射hCG后的6 - 18小时以及成年周期动物发情日早晨的0400时和1000时。在大多数来自大的排卵前卵泡的颗粒细胞和卵丘细胞、构成间质腺组织的大多数细胞以及生长中和成熟卵泡的单个卵泡膜细胞中，均观察到PACAP信使核糖核酸（mRNA）和PACAP免疫反应性。在PMSG/hCG处理的动物中，在PMSG注射后12小时，在单个卵泡膜细胞和间质细胞中也观察到阳性信号。在所检查的所有卵巢中均观察到PACAP免疫反应性神经纤维，它们位于间质腺组织或疏松结缔组织中。卵巢组织的Northern印迹分析证实了PACAP在排卵前和排卵时的短暂表达。观察到三种转录本。最主要的条带大小约为1.2千碱基对，与在睾丸中观察到的转录本相对应。还观察到两条大小分别为2.4和3.0千碱基对的条带，与神经组织的转录本相对应。PACAP阳性细胞和神经纤维的复杂分布表明，该肽可能是卵巢排卵周期以及早期卵泡发生过程中许多事件的局部调节因子。
