Abdel-Ghani M A, El-Sherry T M, Abdelhafeez H H
Department of Theriogenology, Faculty of Veterinary Medicine, Assuit University, Assuit, Egypt.
Department of Anatomy and Histology, Faculty of Veterinary Medicine, Assuit University, Assuit, Egypt.
Reprod Domest Anim. 2016 Oct;51(5):795-803. doi: 10.1111/rda.12753. Epub 2016 Aug 23.
To improve the reproductive performance of water buffalo to level can satisfy our needs, the mechanisms controlling ovarian follicular growth and development should be thoroughly investigated. Therefore, in this study, the expressions of growth differentiation factor-9 (GDF-9) in buffalo ovaries were examined by immunohistochemistry, and the effects of GDF-9 treatment on follicle progression were investigated using a buffalo ovary organ culture system. Frozen-thawed buffalo ovarian follicles within slices of ovarian cortical tissue were cultured for 14 days in the presence or absence of GDF-9. After culture, ovarian slices were fixed, sectioned and stained. The follicles were morphologically analysed and counted. Expression pattern of GDF-9 was detected in oocytes from primordial follicles onwards, besides, also presented in granulosa cells. Moreover, GDF-9 was detected in mural granulosa cells and theca cells of pre-antral follicles. In antral follicles, cumulus cells and theca cells displayed positive expression of GDF-9. In corpora lutea, GDF-9 was expressed in both granulosa and theca lutein cells. After in vitro culture, there was no difference in the number of primordial follicles between cultured plus GDF-9 and cultured control that indicated the GDF-9 treatment has no effect on the primordial to primary follicle transition. GDF-9 treatment caused a significant decrease in the number of primary and secondary follicles compared with controls accompanied with a significant increase in pre-antral and antral follicles. These results suggest that a larger number of primary and secondary follicles were stimulated to progress to later developmental stages when treated with GDF-9. Vitrification/warming of buffalo ovarian tissue had a little remarkable effect, in contrast to culturing for 14 days, on the expression of GDF-9. In conclusion, treatment with GDF-9 was found to promote progression of primary follicle that could provide an alternative approach to stimulate early follicle development and to improve therapies for the most common infertility problem in buffaloes (ovarian inactivity).
为了将水牛的繁殖性能提高到能满足我们需求的水平，必须深入研究控制卵巢卵泡生长和发育的机制。因此，在本研究中，通过免疫组织化学检测了水牛卵巢中生长分化因子9（GDF-9）的表达，并使用水牛卵巢器官培养系统研究了GDF-9处理对卵泡发育进程的影响。将冷冻解冻的水牛卵巢皮质组织切片内的卵泡在有或没有GDF-9的情况下培养14天。培养后，将卵巢切片固定、切片并染色。对卵泡进行形态学分析和计数。从原始卵泡开始，在卵母细胞中检测到GDF-9的表达模式，此外，在颗粒细胞中也有表达。此外，在窦前卵泡的壁颗粒细胞和膜细胞中检测到GDF-9。在窦卵泡中，卵丘细胞和膜细胞显示出GDF-9的阳性表达。在黄体中，GDF-9在颗粒黄体细胞和膜黄体细胞中均有表达。体外培养后，添加GDF-9培养组和对照培养组的原始卵泡数量没有差异，这表明GDF-9处理对原始卵泡向初级卵泡的转变没有影响。与对照组相比，GDF-9处理导致初级和次级卵泡数量显著减少，同时窦前卵泡和窦卵泡数量显著增加。这些结果表明，用GDF-9处理时，大量的初级和次级卵泡被刺激进入后期发育阶段。与培养14天相比，水牛卵巢组织的玻璃化/复温对GDF-9的表达影响不大。总之，发现GDF-9处理可促进初级卵泡的发育进程，这可为刺激早期卵泡发育和改善水牛最常见的不育问题（卵巢无活性）的治疗提供一种替代方法。
