Bruno J B, Celestino J J H, Lima-Verde I B, Lima L F, Matos M H T, Araújo V R, Saraiva M V A, Martins F S, Name K P O, Campello C C, Báo S N, Silva J R V, Figueiredo J R
Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceara, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60.740-000, Brazil.
Reprod Fertil Dev. 2009;21(5):679-87. doi: 10.1071/RD08181.
The aim of the present study was to evaluate the effect of vascular endothelial growth factor (VEGF) on the survival and growth of goat preantral follicles after in vitro culture and to verify the expression of VEGF receptor (VEGFR)-2 in goat ovaries. Ovarian fragments were cultured for 1 or 7 days in minimal essential medium (MEM) with different concentrations of VEGF (1, 10, 50, 100 or 200 ng mL(-1)). Non-cultured (fresh control) and cultured tissues were processed for histological and ultrastructural studies. The results showed that 200 ng mL(-1) VEGF resulted in a similar percentage of normal preantral follicles after 1 and 7 days of culture compared with control. Compared with basic culture medium alone, an increase in follicular and oocyte diameters was observed in the presence of 10 ng mL(-1) VEGF after 7 days culture. Ultrastructural analysis confirmed follicular integrity after 7 days culture in the presence of 200 ng mL(-1) VEGF. Immunohistochemical studies demonstrated the expression of VEGFR-2 in oocytes and granulosa cells of all follicular stages, except in granulosa cells of primordial follicles. In conclusion, the present study has shown that VEGF maintains follicular ultrastructural integrity and promotes follicular growth. In addition, VEGFR-2 is expressed in oocytes of caprine ovarian follicles at all developmental stages and in granulosa cells of developing follicles.
本研究的目的是评估血管内皮生长因子(VEGF)对体外培养后山羊腔前卵泡存活和生长的影响,并验证VEGF受体(VEGFR)-2在山羊卵巢中的表达。将卵巢组织块在含有不同浓度VEGF(1、10、50、100或200 ng mL⁻¹)的最低必需培养基(MEM)中培养1天或7天。对未培养(新鲜对照)和培养后的组织进行组织学和超微结构研究。结果显示,与对照组相比,200 ng mL⁻¹ VEGF在培养1天和7天后导致正常腔前卵泡的比例相似。与单独的基础培养基相比,培养7天后,在10 ng mL⁻¹ VEGF存在的情况下,卵泡和卵母细胞直径增加。超微结构分析证实,在200 ng mL⁻¹ VEGF存在的情况下培养7天后卵泡完整性良好。免疫组织化学研究表明,除原始卵泡的颗粒细胞外,VEGFR-2在所有卵泡阶段的卵母细胞和颗粒细胞中均有表达。总之,本研究表明VEGF维持卵泡超微结构的完整性并促进卵泡生长。此外,VEGFR-2在山羊卵巢卵泡的所有发育阶段的卵母细胞以及发育中卵泡的颗粒细胞中均有表达。