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在体外培养分离的山羊腔前卵泡过程中,卵泡生长加速对卵泡存活和卵母细胞减数分裂恢复具有不利影响。

Accelerated follicle growth during the culture of isolated caprine preantral follicles is detrimental to follicular survival and oocyte meiotic resumption.

作者信息

Apolloni Livia Brunetti, Bruno Jamily Bezerra, Alves Benner Geraldo, Ferreira Anna Clara Accioly, Paes Victor Macêdo, Moreno Jesus de Los Reyes Cadenas, de Aguiar Francisco Léo Nascimento, Brandão Felipe Zandonadi, Smitz Johan, Apgar Gary, de Figueiredo José Ricardo

机构信息

Laboratory of Manipulation of Oocyte Enclosed in Preantral Follicles-LAMOFOPA, Faculty of Veterinary, State University of Ceará, Fortaleza, Brazil.

Department of Animal Reproduction, Faculty of Veterinary, Federal University Fluminense, Niterói, Rio de Janeiro, Brazil.

出版信息

Theriogenology. 2016 Oct 1;86(6):1530-1540. doi: 10.1016/j.theriogenology.2016.05.012. Epub 2016 May 24.

Abstract

This study investigated the effect of androstenedione (A4) alone or in association with different concentrations of bovine recombinant FSH on the IVC of isolated goat preantral follicles. Follicles were mechanically isolated from ovarian tissue and cultured for 18 days in α-minimum essential medium supplemented or not with A4 (10 ng/mL) alone or in association with fixed (A4 + FixFSH: 100 ng/mL) or sequential (A4 + SeqFSH: Day 0, 100 ng/mL; Day 6, 500 ng/mL; Day 12, 1000 ng/mL) concentrations of FSH. After 18 days, the oocytes were recovered for IVM and fluorescence analysis. At Day 18 of culture, only A4 + SeqFSH treatment showed a lower (P < 0.05) rate of intact follicles, survival probability, and meiotic resumption, as well as higher (P < 0.05) percentage of degeneration and/or extrusion after antrum formation. Taken together, these results reported a positive correlation between fast-growing follicles and follicles that degenerated and/or extruded after antrum formation. When compared with control, the addition of A4 alone or in association of FSH did not increase (P > 0.05) the estradiol production or androstenedione levels on Day 6. However, on Day 18, the androstenedione levels were significantly lower in A4 + SeqFSH treatment when compared with A4 alone or to A4 + FixFSH treatments, whereas the estradiol production did not differ (P > 0.05). In summary, this study found that accelerated follicle growth negatively impacted the morphology of caprine preantral follicle cultured in vitro. In addition, the association of androstenedione with increasing concentration of FSH was detrimental to follicular survival and oocyte meiotic resumption.

摘要

本研究调查了雄烯二酮(A4)单独使用或与不同浓度的牛重组促卵泡素(FSH)联合使用对分离的山羊腔前卵泡体外成熟培养(IVC)的影响。卵泡从卵巢组织中机械分离出来,在添加或不添加A4(10 ng/mL)的α-最低必需培养基中培养18天,A4单独使用或与固定浓度(A4 + 固定FSH:100 ng/mL)或序贯浓度(A4 + 序贯FSH:第0天,100 ng/mL;第6天,500 ng/mL;第12天,1000 ng/mL)的FSH联合使用。18天后,回收卵母细胞进行体外成熟培养(IVM)和荧光分析。在培养的第18天,只有A4 + 序贯FSH处理显示完整卵泡率、存活概率和减数分裂恢复率较低(P < 0.05),以及在卵泡腔形成后退化和/或排出的百分比更高(P < 0.05)。综上所述,这些结果表明快速生长的卵泡与卵泡腔形成后退化和/或排出的卵泡之间存在正相关。与对照组相比,单独添加A4或与FSH联合添加在第6天时并未增加(P > 0.05)雌二醇产量或雄烯二酮水平。然而,在第18天,与单独使用A4或A4 + 固定FSH处理相比,A4 + 序贯FSH处理的雄烯二酮水平显著降低,而雌二醇产量没有差异(P > 0.05)。总之,本研究发现卵泡生长加速对体外培养的山羊腔前卵泡形态有负面影响。此外,雄烯二酮与FSH浓度增加联合使用对卵泡存活和卵母细胞减数分裂恢复有害。

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