Gonadotropin-dependent regulation of bovine pituitary adenylate cyclase-activating polypeptide in ovarian follicles prior to ovulation.
Gonadotropin-dependent regulation of bovine pituitary adenylate cyclase-activating polypeptide in ovarian follicles prior to ovulation.
复制标题
排卵前卵泡中牛垂体腺苷酸环化酶激活多肽的促性腺激素依赖性调节。
作者：
K. Sayasith;K. Brown;J. Sirois
为研究牛垂体腺苷环化酶激活多肽(PACAP)在排卵前卵泡中的调控作用，采用RT-PCR方法克隆了PACAP基因。它的开放阅读框(ORF)由531bp组成，编码176个氨基酸的蛋白质，与其他PACAP同源物有76-90%的同源性。以人绒毛膜促性腺激素(HCG)处理后0~24 h的牛排卵前卵泡为研究对象，半定量RT-PCR/Southern印迹分析结果表明，PACAP m RNA水平在0 h较低，6 h和12 h显著升高(P<；0.05)，hCG后18 h和24 h下降。PACAP在牛脑垂体、睾丸、肠道和子宫中的表达水平较高，而在其他组织中的表达水平较低。对分离的颗粒细胞和膜细胞的分析表明，hCG处理后，两种细胞中PACAP的转录均显著增加，而原代颗粒细胞培养显示，Forsklin处理后，PACAP及其受体PAC-1和VPAC-2mRNA水平较高。蛋白激酶A催化亚单位(PKA)的过度表达被刺激，但H89或PKA抑制蛋白处理抑制PACAP的mRNA表达，而PACAP过表达则刺激PGHS-2、PGES、EP2受体、孕激素受体和ADAMTS-1的转录丰度增加，但不促进P450侧链断裂和P450芳香化酶的表达。因此，本研究证实了促性腺激素对牛排卵前卵泡中PACAP基因表达的依赖调节，PKA激活在颗粒细胞PACAP表达中的重要性，以及PACAP在排卵过程中对基因表达的刺激作用。
To study the regulation of bovine pituitary adenylate cyclase-activating polypeptide (PACAP) in preovulatory follicles prior to ovulation, PACAP cDNA was isolated by RT-PCR. Its open reading frame (ORF) is composed of 531 bp, and encodes for a 176-amino acid protein that bears 76-90% identity with other PACAP homologs. Using bovine preovulatory follicles obtained between 0 and 24 h after human chorionic gonadotropin (hCG) and semiquantitative RT-PCR/Southern blot, we demonstrate that levels of PACAP mRNA were low at 0 h, markedly increased at 6 and 12 h (P<0.05), and declined 18 and 24 h after hCG. Levels of PACAP mRNA were high in the bovine pituitary, testis, intestine and uterus, but moderate to low in other tissues. Analyses performed on isolated preparations of granulosa and theca cells showed a significant increase of PACAP transcripts in both cell types after hCG, whereas primary granulosa cell cultures revealed high levels of PACAP as well as its receptors PAC-1 and VPAC-2 mRNA after forskolin treatment. Overexpression of the catalytic subunit of protein kinase A (PKA) in granulosa cells stimulated, but treatment with H89 or PKA inhibitor protein inhibited PACAP mRNA expression, whereas PACAP overexpression stimulated an increase in abundance of transcripts for PGHS-2, PGES, EP2 receptor, progesterone receptor, and ADAMTS-1, but not for P450-side chain cleavage and P450 aromatase. Thus, this study demonstrates the gonadotropin-dependent regulation of PACAP mRNA in bovine preovulatory follicles, the importance of PKA activation in the expression of PACAP in granulosa cells, and stimulating effect of PACAP on gene expression during the ovulatory process.
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DOI：
10.1073/pnas.080073497
发表时间：
2000-04-25
影响因子：
11.1
作者：
Robker, RL;Russell, DL;Richards, JS
通讯作者：
Richards, JS
垂体腺苷酸环化酶激活多肽刺激培养的大鼠颗粒细胞中的类固醇生成和腺苷 3,5-单磷酸积累。
影响因子：
4.8
作者：
Zhong,Y;Kasson,BG
通讯作者：
Kasson,BG
影响因子：
21.1
作者：
D. Vaudry;B. Gonzalez;M. Basille;L. Yon;A. Fournier;H. Vaudry
通讯作者：
D. Vaudry;B. Gonzalez;M. Basille;L. Yon;A. Fournier;H. Vaudry
DOI：
10.1016/0006-291x(89)91757-9
发表时间：
1989-10-16
影响因子：
3.1
作者：
MIYATA, A;ARIMURA, A;COY, DH
通讯作者：
COY, DH
影响因子：
3.5
作者：
Boerboom, D;Russell, DL;Sirois, J
通讯作者：
Sirois, J