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BMPRIB 和 BMPRII 在山羊卵巢卵泡中的 mRNA 表达水平,以及 BMP-15 对腔前卵泡发育的体外作用。

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

机构信息

Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil.

出版信息

Cell Tissue Res. 2012 Apr;348(1):225-38. doi: 10.1007/s00441-012-1361-4. Epub 2012 Mar 2.

Abstract

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.

摘要

本研究评估了骨形态发生蛋白受体 BMPRIB 和 BMPRII mRNA 在山羊卵泡中的水平,以及骨形态发生蛋白-15(BMP-15)对体外培养的原始卵泡发育的影响。实时聚合酶链反应(PCR)用于分析山羊原始卵泡和小、大腔卵泡中 BMPRIB 和 BMPRII mRNA 的水平。从小山羊卵巢中分离原始卵泡(≥150 μm),并在添加或不添加 BMP-15(10、50 或 100ng/ml)的α-MEM(+)中培养 18 天。培养结束时,一些卵泡固定用于超微结构评估。实时 PCR 显示 BMPRII mRNA 水平从初级卵泡到次级卵泡降低。大腔卵泡的颗粒细胞/膜细胞中 BMPRIB mRNA 水平较高,与小腔卵泡相比。此外,大腔卵泡的卵丘-卵母细胞复合体中 BMPRII mRNA 的表达程度大于其各自的颗粒细胞/膜细胞。在培养中,50ng/ml 的 BMP-15 可在 18 天后积极影响腔形成和卵泡生长,并维持卵泡完整性。因此,BMPRIB 和 BMPRII mRNA 存在于所有卵泡类别中。BMP-15(50ng/ml)刺激 18 天培养后分离山羊原始卵泡的生长、腔形成和超微结构完整性。

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