Transient periovulatory expression of pituitary adenylate cyclase activating peptide in rat ovarian cells
Transient periovulatory expression of pituitary adenylate cyclase activating peptide in rat ovarian cells
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DOI：
10.1210/en.137.11.4779
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发表时间：
1996-11-01
期刊：
影响因子：
4.8
通讯作者：
Fahrenkrug, J
中科院分区：
文献类型：
--
作者：
Gras, S;Hannibal, J;Fahrenkrug, J
垂体腺苷酸环化酶激活多肽（垂体腺苷酸环化酶激活多肽，PACAP）是一种有效的cAMP形成刺激物，广泛分布于中枢神经系统和一些外周器官的神经元元件中。最近，PACAP也被证明在大鼠睾丸的生精细胞中有分期特异性表达。这促使我们通过地高西格素utp标记的RNA探针原位杂交和使用特异性单克隆抗体的免疫组织化学来检测PACAP在大鼠卵巢发情周期中的细胞特异性表达。在PMSG处理后0、12、24、48、54、57、58.5、60、61.5、63、66、72和96 h，对PMS促性腺激素(PMSG)/人促性腺激素（hCG）处理的未成熟动物卵巢进行检测。在发情周期的每天1000 h和发情早晨0400 h检查成年周期大鼠的卵巢。在PMSG诱导周期中hCG注射后6-18 h的排卵期和成体周期动物发情早晨0400 h和1000 h，均观察到PACAP阳性细胞。PACAP信使RNA （mRNA）和PACAP免疫反应性在大排卵泡的颗粒细胞和积云细胞、大部分间质腺组织细胞以及生长和成熟卵泡的孤立膜细胞中均有表达。在PMSG/hCG处理的动物中，注射PMSG后12 h，单独的卵泡膜和间质细胞也观察到阳性信号。所有卵巢均可见PACAP免疫反应性神经纤维，位于间质腺组织或疏松结缔组织。卵巢组织的Northern印迹证实了短暂的排卵期和排卵期前PACAP的表达。观察到三个转录本。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2.4和3.0千碱基对的两个条带，对应于神经组织的转录本。PACAP阳性细胞和神经纤维的复杂分布表明，该肽可能是发生在卵巢排卵期和早期卵泡发生期间的许多事件的局部调节剂。
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 gonadatropin (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.