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Activin promotes oocyte development in ovine preantral follicles in vitro

机译:激活素体外促进绵羊前窦卵泡的卵母细胞发育

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摘要

Activins have been implicated as important regulating factors for many reproductive processes. The aim of this study was to determine the effect of activin A on the development of ovine preantral follicles in vitro. Mechanically isolated preantral follicles (161 ± 2 microm) were cultured for 6 days in the presence of human recombinant activin A (0, 10 and 100 ng/ml). Half of the medium was replaced every second day and follicle diameters were measured. Conditioned medium was subsequently analysed for oestradiol content using a delayed enhancement lanthanide fluorometric immunoassay (DELFIA). At the end of the culture period, follicles were fixed and processed for histology, after which oocyte diameter and granulosa cell death were measured. There was significant follicle growth over 6 days in all groups (p < 0.001). Activin, at both concentrations, increased follicle growth over control levels by Day 6 (p < 0.05). Oocyte diameters were also significantly increased by Day 6 of culture in all groups (p < 0.05), with 100 ng/ml activin increasing oocyte diameter over control levels (p < 0.05). Activin, at both concentrations, increased oestradiol production on Day 2 of culture, but this increase was not sustained during the culture period. Moreover, activin did not have any effect on antrum formation or follicle survival. In conclusion, activin promoted ovine preantral follicle and oocyte growth in vitro, but did not accelerate follicle differentiation over a six-day culture period. These results support a paracrine role for activin A during early oocyte and follicular development.
机译:激活素已被认为是许多生殖过程的重要调节因子。这项研究的目的是确定激活素A在体外对绵羊窦前卵泡发育的影响。在人重组激活素A(0、10和100 ng / ml)的存在下,将机械分离的窦前卵泡(161±2微米)培养6天。每隔两天更换一半的培养基,并测量卵泡直径。随后使用延迟增强镧系元素荧光免疫分析(DELFIA)分析条件培养基中的雌二醇含量。培养结束时,将卵泡固定并进行组织学处理,然后测量卵母细胞直径和颗粒细胞死亡。所有组在6天内都有明显的卵泡生长(p <0.001)。到第6天,两种浓度的激活素都比对照水平增加了卵泡生长(p <0.05)。到培养的第6天,所有组的卵母细胞直径也显着增加(p <0.05),其中100 ng / ml激活素使卵母细胞直径超过对照水平(p <0.05)。两种浓度下的激活素在培养的第2天都增加了雌二醇的产生,但在培养期间并未持续这种增加。此外,激活素对胃窦的形成或卵泡的存活没有任何影响。总之,激活素在体外可促进绵羊窦前卵泡和卵母细胞的生长,但在六天的培养期内并未促进卵泡的分化。这些结果支持了早期卵母细胞和卵泡发育过程中激活素A的旁分泌作用。

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