Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceara, Fortaleza, Brazil.
Cells Tissues Organs. 2010;191(5):414-21. doi: 10.1159/000272317. Epub 2009 Dec 24.
The aim of this study was to evaluate the effect of vasoactive intestinal peptide (VIP)on the survival, activation and growth of goat preantral follicles after in vitro culture. The ovarian cortex was divided into small pieces and one fragment was immediately fixed (control). The remaining fragments were cultured in vitro for 1 or 7 days at 39 degrees C and 5% CO(2), in supplemented minimum essential medium (MEM(+)) with or without different concentrations of VIP (1, 10, 50, 100 or 200 ng/ml). Noncultured (fresh control) and cultured ovarian fragments were processed for histological analysis and transmission electron microscopy. Follicles were classified as primordial or developing, and as normal or degenerated. Our findings indicate that when compared with control, addition of all concentrations of VIP except 200 ng/ml resulted in similar percentages of normal preantral follicles after 1 and 7 days of culture. Culture of ovarian cortex tissue for 1 and 7 days increased the percentage of follicular activation in all treatments when compared with control, except with 1 ng/ml of VIP after 1 day. However, no difference was observed between VIP-treated and MEM(+)-treated follicles. In addition, after 7 days of culture, the highest follicular and oocyte diameters were observed in follicles cultured with 10 ng/ml VIP relative to MEM(+) alone. Transmission electron microscopy showed ultrastructural integrity of follicles after 7 days of culture in 10 ng/ml VIP. In conclusion, this study demonstrates that VIP maintains follicular integrity and stimulates caprine preantral follicle growth.
本研究旨在评估血管活性肠肽(VIP)对体外培养山羊腔前卵泡存活、激活和生长的影响。将卵巢皮质切成小块,其中一个片段立即固定(对照)。其余片段在 39°C 和 5%CO2 下,于添加有或无不同浓度 VIP(1、10、50、100 或 200ng/ml)的补充基本最小培养基(MEM(+))中体外培养 1 或 7 天。非培养(新鲜对照)和培养的卵巢片段进行组织学分析和透射电镜检查。卵泡被分类为原始卵泡或发育卵泡,并分为正常卵泡或退化卵泡。我们的研究结果表明,与对照相比,除 200ng/ml 外,所有浓度的 VIP 均导致培养 1 和 7 天后正常腔前卵泡的比例相似。与对照相比,培养卵巢皮质组织 1 和 7 天可增加所有处理中卵泡激活的百分比,除了 1 天的 1ng/ml VIP。然而,在 VIP 处理和 MEM(+)处理的卵泡之间未观察到差异。此外,在培养 7 天后,与单独使用 MEM(+)相比,在 10ng/ml VIP 中培养的卵泡中观察到最高的卵泡和卵母细胞直径。透射电镜显示,在 10ng/ml VIP 中培养 7 天后,卵泡具有完整的超微结构。总之,本研究表明 VIP 维持卵泡的完整性并刺激山羊腔前卵泡的生长。