Laboratory of Manipulation of Oocytes and Preantral Follicles, State University of Ceará, Fortaleza, CE, Brazil.
Reprod Sci. 2011 Dec;18(12):1222-9. doi: 10.1177/1933719111410709. Epub 2011 Jun 21.
The aims of this study were to evaluate the expression of keratinocyte growth factor (KGF) in goat ovaries and to study its effects on preantral follicle survival and development. The ovaries were used for immunohistochemistry or for in vitro culture for 1 or 7 days with KGF (0, 1, 10, 50, 100, 150, or 200 ng/mL). Noncultured (fresh control) and cultured ovarian slices were processed for histological analysis and transmission electron microscopy (TEM). The results showed that after 7 days of in vitro culture, all treatments had a significant reduction in the percentage of normal follicles compared with the fresh control. After 7 days of culture, the highest KGF concentrations (150 and 200 ng/mL) induced a significant reduction in the percentage of normal follicles compared with the tissues cultured in the absence (α-MEM(+) alone) or presence of 1, 10, and 50 ng/mL KGF. Transmission electron microscopy confirmed follicular integrity after 7 days of culture in 1 ng/mL KGF. In addition, compared with the fresh control, the percentage of growing follicles was significantly increased in all treatments after 1 or 7 days of culture. Immunohistochemical analyses showed the expression of KGF in oocytes and granulosa cells in all follicle developmental stages as well as in thecal and stromal cells. In conclusion, this study demonstrated that, at the lowest concentration (1 ng/mL), KGF maintained the ultrastructure of goat preantral follicles cultured in vitro for up to 7 days. Furthermore, the KGF protein was widely distributed in goat ovaries, especially in ovarian follicles.
本研究旨在评估角质细胞生长因子(KGF)在山羊卵巢中的表达,并研究其对原始卵泡存活和发育的影响。将卵巢用于免疫组织化学或体外培养 1 或 7 天,用 KGF(0、1、10、50、100、150 或 200ng/ml)处理。未经培养(新鲜对照)和培养的卵巢切片进行组织学分析和透射电镜(TEM)处理。结果显示,与新鲜对照组相比,体外培养 7 天后,所有处理组正常卵泡的比例均显著降低。培养 7 天后,与在无 KGF(仅α-MEM(+))或存在 1、10 和 50ng/ml KGF 时培养的组织相比,最高浓度(150 和 200ng/ml)的 KGF 显著降低了正常卵泡的比例。透射电镜证实,在 1ng/ml KGF 培养 7 天后,卵泡仍保持完整。此外,与新鲜对照组相比,培养 1 或 7 天后,所有处理组的生长卵泡比例均显著增加。免疫组织化学分析显示,在所有卵泡发育阶段,包括卵母细胞和颗粒细胞以及间质和基质细胞中均表达 KGF。综上所述,本研究表明,在最低浓度(1ng/ml)下,KGF 可维持山羊原始卵泡在体外培养长达 7 天的超微结构。此外,KGF 蛋白在山羊卵巢中广泛分布,尤其是在卵巢卵泡中。