Shi Fangxiong, Stewart Robert L, Perez Emerson, Chen Jean Y-H, LaPolt Philip S
Department of Biological Sciences, California State University-Los Angeles, Los Angeles, California 90032, USA.
Biol Reprod. 2004 Jun;70(6):1552-61. doi: 10.1095/biolreprod.103.025510. Epub 2004 Jan 28.
Soluble guanylyl cyclase (sGC) is activated by nitric oxide (NO) and carbon monoxide, resulting in cGMP production. Recent studies indicate that NO and cGMP influence ovarian functions. However, little information is available regarding the ovarian expression of sGC. The present study examined sGC alpha(1) and beta(1) subunit protein levels in the ovary during postnatal development, gonadotropin-induced follicle growth, ovulation, and luteinization as well as in cultured rat granulosa cells. In postnatal rats, sGC alpha(1) subunit immunoreactivity was high in granulosa cells of primordial and primary follicles on Day 5 but low in granulosa cells of larger follicles on Days 10 and 19. Theca cells of developing follicles, but not stromal cells, also demonstrated moderate sGC alpha(1) immunoreactivity. In gonadotropin- treated immature rats, intense sGC alpha(1) subunit staining was similarly observed in granulosa cells of primordial and primary follicles, but such staining was low in granulosa cells of small antral follicles and undetectable in granulosa cells of large antral and preovulatory follicles. Following ovulation, corpora lutea expressed moderate sGC alpha(1) immunoreactivity. Similar ovarian localization and expression patterns were seen for sGC beta(1), indicating regulated coexpression of sGC subunits. Immunoblot analysis revealed no change in total ovarian sGC alpha(1) and beta(1) subunit protein levels during gonadotropin treatment. Similarly, no effect of FSH on sGC subunit protein levels was apparent in cultured granulosa cells. These findings indicate regulated, cell- specific patterns of sGC expression in the ovary and are consistent with roles for cGMP in modulating ovarian functions.
可溶性鸟苷酸环化酶(sGC)可被一氧化氮(NO)和一氧化碳激活,从而产生环磷酸鸟苷(cGMP)。最近的研究表明,NO和cGMP会影响卵巢功能。然而,关于sGC在卵巢中的表达情况,目前所知甚少。本研究检测了出生后发育过程中、促性腺激素诱导的卵泡生长、排卵、黄体化过程中卵巢内sGCα(1)和β(1)亚基的蛋白水平,以及培养的大鼠颗粒细胞中的表达情况。在出生后的大鼠中,出生第5天原始卵泡和初级卵泡的颗粒细胞中sGCα(1)亚基的免疫反应性较高,但在出生第10天和第19天较大卵泡的颗粒细胞中则较低。发育卵泡的膜细胞,而非基质细胞,也显示出中等程度的sGCα(1)免疫反应性。在促性腺激素处理的未成熟大鼠中,原始卵泡和初级卵泡的颗粒细胞中同样观察到强烈的sGCα(1)亚基染色,但在小窦状卵泡的颗粒细胞中这种染色较低,而在大窦状卵泡和排卵前卵泡的颗粒细胞中则无法检测到。排卵后,黄体表达中等程度的sGCα(1)免疫反应性。sGCβ(1)也呈现出类似的卵巢定位和表达模式,表明sGC亚基存在协同表达调控。免疫印迹分析显示,促性腺激素处理期间卵巢中sGCα(1)和β(1)亚基的总蛋白水平没有变化。同样,在培养的颗粒细胞中,促卵泡激素(FSH)对sGC亚基蛋白水平也没有明显影响。这些发现表明卵巢中sGC的表达存在调控的细胞特异性模式,并且与cGMP在调节卵巢功能中的作用相一致。