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BMPRIB and BMPRII mRNA expression levels in goat ovarian follicles and the in vitro effects of BMP-15 on preantral follicle development

机译:山羊卵巢卵泡中BMPRIB和BMPRII mRNA的表达水平以及BMP-15对窦前卵泡发育的体外影响

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This study evaluated the levels of bone morphogenetic protein receptors BMPRIB and BMPRII mRNA in goat follicles and the effects of bone morphogenetic protein-15 (BMP-15) on the in vitro development of cultured preantral follicles. Real-time polymerase chain reaction (PCR) was used to analyze the levels of BMPRIB and BMPRII mRNA in caprine preantral follicles and in small and large antral follicles. Preantral follicles (≥150 μm) were also isolated from goat ovaries and cultured for 18 days in α-MEM+ supplemented with or without BMP-15 (10, 50, or 100 ng/ml). At the end of culture, some follicles were fixed for ultrastructural evaluation. Real-time PCR showed a reduction in BMPRII mRNA levels from the primary to secondary follicles. Higher levels of BMPRIB mRNA were observed in granulosa/theca cells from large antral follicles compared with small antral follicles. Moreover, BMPRII mRNA was expressed to a greater extent in cumulus–oocyte complexes from large antral follicles than in their respective granulosa/theca cells. In culture, 50 ng/ml BMP-15 positively influenced antral cavity formation and follicle growth after 18 days and also maintained follicular integrity. Thus, BMPRIB and BMPRII mRNAs are present in all follicular categories. BMP-15 (50 ng/ml) stimulates growth, antrum formation and the ultrastructural integrity of isolated caprine preantral follicles after 18 days of culture.
机译:这项研究评估了山羊卵泡中骨形态发生蛋白受体BMPRIB和BMPRII mRNA的水平,以及骨形态发生蛋白15(BMP-15)对体外培养的腔前卵泡发育的影响。实时聚合酶链反应(PCR)用于分析山羊窦前卵泡和大,小肛门窦卵泡中BMPRIB和BMPRII mRNA的水平。还从山羊卵巢中分离出窦前卵泡(≥150μm),并在补充或不补充BMP-15(10、50或100 ng / ml)的α-MEM + 中培养18天。培养结束时,固定了一些卵泡以进行超微结构评估。实时PCR显示从初级卵泡到次级卵泡的BMPRII mRNA水平降低。与小窦卵泡相比,在大窦卵泡的颗粒/皮膜细胞中观察到更高水平的BMPRIB mRNA。此外,相比于它们各自的颗粒/卵泡膜细胞,来自大型肛门卵泡的卵-卵母细胞复合物中BMPRII mRNA的表达程度更高。在培养中,50 ng / ml BMP-15对18天后的肛门腔形成和卵泡生长有积极影响,并保持卵泡完整性。因此,BMPRIB和BMPRII mRNA存在于所有卵泡类别中。培养18天后,BMP-15(50 ng / ml)刺激生长,窦腔形成和分离的山羊窦前卵泡超微结构完整性。

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