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山羊卵巢中编码血小板衍生生长因子(PDGF)受体的mRNA表达水平及PDGF对山羊腔前卵泡体外发育的影响

Expression levels of mRNA-encoding PDGF receptors in goat ovaries and the influence of PDGF on the in vitro development of caprine pre-antral follicles.

作者信息

Brito I R, Lima I M T, Saraiva M V A, Silva C M G, Magalhães-Padilha D M, Araújo V R, Barreto Luz V, Barbalho Silva A W, Campello C C, Silva J R V, Figueiredo J R

机构信息

Faculty of Veterinary, Laboratory of Manipulation of Oocytes and Preantral Follicles, PPGCV, State University of Ceará, Fortaleza, CE, Brazil.

出版信息

Reprod Domest Anim. 2012 Oct;47(5):695-703. doi: 10.1111/j.1439-0531.2011.01946.x. Epub 2011 Dec 3.

Abstract

The aims of this study were to investigate the expression levels of mRNA for platelet-derived growth factor (PDGF) receptors (PDGFR-α and -β) in caprine follicles at different developmental stages and to evaluate the influence of PDGF on the in vitro development of pre-antral follicles. For this, goat primordial, primary and secondary follicles, as well as small (1-3 mm) and large (3-6 mm) antral follicles, were obtained, and PDGFR-α and -β mRNA levels were quantified by real-time PCR. Furthermore, pre-antral follicles (≥ 200 μm) were isolated from goat ovaries and cultured for 18 days in α- minimum essential medium supplemented with PDGF at 50 or 100 ng/ml, containing or not FSH. Real-time PCR showed highest PDGFR-α mRNA levels in secondary follicles, while PDGFR-β mRNA levels were highest in primary follicles onwards. Both receptors showed higher mRNA levels in granulosa/theca cells from small and large antral follicles than in their corresponding cumulus-oocyte complexes. In culture, the percentage of antrum formation was significantly higher in 100 ng/ml PDGF compared with the same PDGF concentration associated with FSH. After 18 days, PDGF in both concentrations associated with FSH promoted follicular growth significantly higher than the control. Moreover, the addition of FSH to 50 ng/ml PDGF positively influenced the follicular growth when compared with the same PDGF concentration in the absence of FSH. In conclusion, PDGF is important for early goat folliculogenesis, because the presence of PDGFR-α and -β mRNA was detected in all follicular categories, and PDGF associated with FSH stimulated the growth of goat pre-antral follicles isolated and cultured in vitro.

摘要

本研究的目的是调查不同发育阶段山羊卵泡中血小板衍生生长因子(PDGF)受体(PDGFR-α和-β)的mRNA表达水平,并评估PDGF对体外前体卵泡发育的影响。为此,获取了山羊原始卵泡、初级卵泡和次级卵泡,以及小(1-3毫米)和大(3-6毫米)的有腔卵泡,并通过实时PCR定量PDGFR-α和-β的mRNA水平。此外,从山羊卵巢中分离出前体卵泡(≥200微米),并在添加了50或100纳克/毫升PDGF的α-最低必需培养基中培养18天,培养基中含有或不含有促卵泡激素(FSH)。实时PCR显示,次级卵泡中PDGFR-α的mRNA水平最高,而从初级卵泡开始,PDGFR-β的mRNA水平最高。两种受体在小和大的有腔卵泡的颗粒细胞/卵泡膜细胞中的mRNA水平均高于其相应的卵丘-卵母细胞复合体。在培养中,与相同PDGF浓度联合FSH相比,100纳克/毫升PDGF组的卵泡腔形成百分比显著更高。18天后,与FSH联合的两种浓度的PDGF均显著促进卵泡生长,高于对照组。此外,与不添加FSH的相同PDGF浓度相比,向50纳克/毫升PDGF中添加FSH对卵泡生长有积极影响。总之,PDGF对山羊早期卵泡发生很重要,因为在所有卵泡类别中均检测到PDGFR-α和-β的mRNA,并且与FSH联合的PDGF刺激了体外分离和培养的山羊前体卵泡的生长。

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