Centre for Integrative Physiology, Hugh Robson Building, University of Edinburgh, George Square, Edinburgh EH8 9XD, UK.
Mol Hum Reprod. 2010 Sep;16(9):644-53. doi: 10.1093/molehr/gaq021. Epub 2010 Mar 4.
The aim of this study was to determine the individual and combined effect of activin and follicle stimulating hormone (FSH) on somatic and germ cell development in cultured pre-antral follicles. Pre-antral bovine follicles (mean diameter 157 +/- 3, range 132-199 microm) were cultured for 8 days in serum-free medium in the presence of either 100 ng/ml of recombinant human activin A (rhAct A), 100 ng/ml rhAct A combined with a high (100 ng/ml) or low (50 ng/ml) concentration of recombinant FSH (rFSH) or 50 ng/ml rFSH alone. Intrafollicular connexin 43 expression and actin-based cell adhesion were assessed on Day 2 and 4 of culture. Steroidogenesis was evaluated after Day 4 and 8. Follicles exposed to 100 ng/ml activin maintained expression of connexin 43 at the follicular periphery. In the presence of activin, with or without 100 ng/ml or 50 ng/ml FSH, follicles were steroidogenic undergoing significant growth (P < 0.01), granulosa cell proliferation (P < 0.01) and antral cavity formation (P < 0.05) compared with cultured controls. Maximum oocyte growth occurred in the presence of 100 ng/ml activin alone with a significant percentage of these oocytes maintaining normal morphology over controls (P < 0.05). These results are consistent with a role for activin in maintaining oocyte granulosa cell interactions due to increased peripheral granulosa cell adhesion to the basement membrane and retention of adhesion at the surface of the zona pellucida. Thus, the polarized expression of cell contact interactions promoted by activin supports ongoing folliculogenesis.
本研究旨在确定激活素和卵泡刺激素 (FSH) 对体外培养的前腔前卵泡的体和生殖细胞发育的单独和联合作用。在无血清培养基中培养前腔前牛卵泡(平均直径 157 +/- 3,范围 132-199 微米)8 天,在 100ng/ml 重组人激活素 A (rhAct A)、100ng/ml rhAct A 与高(100ng/ml)或低(50ng/ml)浓度重组 FSH (rFSH) 或 50ng/ml rFSH 单独存在的情况下培养。在培养的第 2 天和第 4 天评估卵泡内连接蛋白 43 的表达和基于肌动蛋白的细胞粘附。在第 4 天和第 8 天评估类固醇生成。暴露于 100ng/ml 激活素的卵泡在卵泡周围维持连接蛋白 43 的表达。在激活素存在的情况下,无论是否存在 100ng/ml 或 50ng/ml FSH,与培养对照组相比,卵泡均具有类固醇生成能力,发生显著生长(P < 0.01)、颗粒细胞增殖(P < 0.01)和腔形成(P < 0.05)。在单独存在 100ng/ml 激活素的情况下,最大卵子生长发生,与对照组相比,这些卵子中有很大比例保持正常形态(P < 0.05)。这些结果与激活素在维持卵母细胞颗粒细胞相互作用中的作用一致,这是由于外围颗粒细胞与基膜的粘附增加以及在透明带表面保持粘附。因此,激活素促进的细胞接触相互作用的极化表达支持持续的卵泡发生。