Silva J R V, van den Hurk R, van Tol H T A, Roelen B A J, Figueiredo J R
Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
J Endocrinol. 2004 Nov;183(2):405-15. doi: 10.1677/joe.1.05756.
We studied the protein and mRNA expression of activin-A, follistatin and activin receptors in goat ovaries to find evidence of their possible role in ovarian activity, particularly in the various stages of follicle development. Ovaries of cyclic goats were collected and then either fixed in paraformaldehyde for immunohistochemical localisation of activin-A, follistatin, activin receptors IIA/B (ActR-IIA/B) and IA (ActR-IA) proteins or used to obtain samples to demonstrate mRNA expression of activin-A (betaA subunit), follistatin, ActR-IIA, -IIB, -IA and -IB, using RT-PCR. For this latter goal, primordial, primary and secondary follicles were isolated mechanically, washed to remove the stromal cells and then used for RT-PCR. In addition, oocytes, cumulus, mural granulosa and theca cells from small (<3 mm) and large (3-6 mm) antral follicles, luteal cells and surface epithelium were collected to study mRNA expression. Activin-A and follistatin proteins were found in oocytes of all follicle classes, granulosa cells from the primary follicle stage onwards, theca cells of antral follicles, corpora lutea and ovarian surface epithelium. In antral follicles, these proteins were detected both in cumulus and mural granulosa cells. ActR-IIA/B protein was found at the same follicular sites, and also in granulosa cells of primordial follicles onward. The localisation of ActR-IA corresponded with that of ActR-IIA/B, but the former protein was absent in the theca of large antral follicles. The mRNAs for activin-A (betaA subunit), follistatin, and ActR-IIA, -IIB, -IA and -IB were detected at all follicular and cellular types studied, except that ActR-IIB was not found in follicles that had not developed an antrum yet. In conclusion, in goat ovaries, transcripts of activin-A (betaA subunit), its receptors and its binding protein follistatin are expressed and their proteins formed at all follicular stages and in corpora lutea. These findings indicate a role of activin-A in the local regulatory system during the entire follicular development and during luteal activity.
我们研究了山羊卵巢中激活素 -A、卵泡抑素及激活素受体的蛋白质和mRNA表达,以探寻它们在卵巢活动,尤其是卵泡发育各个阶段中可能发挥作用的证据。收集处于发情周期的山羊卵巢,随后一部分用多聚甲醛固定,用于激活素 -A、卵泡抑素、激活素受体IIA/B(ActR-IIA/B)和IA(ActR-IA)蛋白的免疫组织化学定位;另一部分用于获取样本,采用逆转录聚合酶链反应(RT-PCR)来检测激活素 -A(βA亚基)、卵泡抑素、ActR-IIA、-IIB、-IA和 -IB的mRNA表达。为实现后一目标,机械分离原始卵泡、初级卵泡和次级卵泡,冲洗以去除基质细胞,然后用于RT-PCR。此外,收集来自小(<3 mm)和大(3 - 6 mm)窦状卵泡的卵母细胞、卵丘、壁层颗粒细胞和卵泡膜细胞、黄体细胞及表面上皮细胞,以研究mRNA表达。在所有卵泡类型的卵母细胞、初级卵泡阶段起的颗粒细胞、窦状卵泡的卵泡膜细胞、黄体及卵巢表面上皮中均发现了激活素 -A和卵泡抑素蛋白。在窦状卵泡中,这些蛋白在卵丘和壁层颗粒细胞中均有检测到。ActR-IIA/B蛋白在相同的卵泡部位被发现,在原始卵泡及之后阶段的颗粒细胞中也有表达。ActR-IA的定位与ActR-IIA/B一致,但在大的窦状卵泡的卵泡膜中不存在前一种蛋白。在所研究的所有卵泡和细胞类型中均检测到了激活素 -A(βA亚基)、卵泡抑素以及ActR-IIA、-IIB、-IA和 -IB的mRNA,但尚未在未形成窦腔的卵泡中发现ActR-IIB。总之,在山羊卵巢中,激活素 -A(βA亚基)、其受体及其结合蛋白卵泡抑素的转录本均有表达,且其蛋白在所有卵泡阶段及黄体中均有形成。这些发现表明激活素 -A在整个卵泡发育及黄体活动期间的局部调节系统中发挥作用。