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卵泡大小对青春期前猪卵丘卵母细胞复合体体外成熟的影响

Effect of Follicle Size on In Vitro Maturation of Pre-Pubertal Porcine Cumulus Oocyte Complexes.

作者信息

Töpfer D, Ebeling S, Weitzel J M, Spannbrucker A C

机构信息

Department of Reproductive Biology, University of Veterinary Medicine Hannover Foundation, Hannover, Germany.

Unit of Reproductive Biology, Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

出版信息

Reprod Domest Anim. 2016 Jun;51(3):370-7. doi: 10.1111/rda.12688. Epub 2016 Apr 3.

Abstract

Very small follicles (<3.0 mm diameter) are over-represented on the surface of ovaries of non-cycling pigs, and the oocytes collected from these follicles generally have reduced developmental competence in vitro. This study examined the effect of follicle size on the nuclear maturation (n = 608), the potential of parthenogenetic activation (n = 243) and the cyclic AMP (cAMP) content of pre-pubertal porcine oocytes (n = 480). In addition, the influence of follicle size on steroid hormone synthesis was analysed. Cumulus oocyte complexes (COCs) flushed from small (2.5-4.0 mm) or large (4.5-6.0 mm) ovarian follicles were cultured for 0, 28 and 46 h. After 46 h of IVM, a greater proportion of oocytes from 4.5- to 6.0-mm follicles reach metaphase II (MII) compared with those from follicles with 2.5-4.0 mm of diameter (96.1 vs 77.0%, respectively; p < 0.001). Parthenogenetic activation of oocytes from large follicles produced higher developmental rates than oocytes from large follicles (p < 0.05). At 28 h, the IVM medium with oocytes from large follicles contained significantly more 17ß-oestradiol (E2 ) than the medium with oocytes from small follicles (5.55 vs 3.45 ng/ml, respectively; p < 0.05) and at 46 h, the medium with oocytes from small follicles contained significantly more progesterone (P4 ) than the medium with oocytes from large follicles (276.7 vs 108.2 ng/ml, respectively, p < 0.05). Porcine oocytes from large follicles have higher nuclear and cytoplasmic maturation capacities, but the differences did not appear to be cAMP-mediated. Our findings also suggest that COCs from small follicles undergo more intensive luteinization than COCs from large follicles. The results show that oocytes from follicles with a diameter greater than 4.0 mm are more suitable for in vitro studies.

摘要

在非发情周期的猪卵巢表面,非常小的卵泡(直径<3.0毫米)占比过高,从这些卵泡中采集的卵母细胞通常在体外的发育能力会降低。本研究考察了卵泡大小对青春期前猪卵母细胞的核成熟(n = 608)、孤雌激活潜力(n = 243)以及环磷酸腺苷(cAMP)含量(n = 480)的影响。此外,还分析了卵泡大小对类固醇激素合成的影响。从小(2.5 - 4.0毫米)或大(4.5 - 6.0毫米)卵巢卵泡中冲洗出的卵丘卵母细胞复合体(COCs)分别培养0、28和46小时。体外成熟培养46小时后,直径4.5 - 6.0毫米卵泡中的卵母细胞达到中期II(MII)的比例高于直径2.5 - 4.0毫米卵泡中的卵母细胞(分别为96.1%和77.0%;p < 0.001)。大卵泡来源的卵母细胞孤雌激活后的发育率高于小卵泡来源的卵母细胞(p < 0.05)。在28小时时,含有大卵泡卵母细胞的体外成熟培养液中17β - 雌二醇(E2)含量显著高于含有小卵泡卵母细胞的培养液(分别为5.55和3.45纳克/毫升;p < 0.05),而在46小时时,含有小卵泡卵母细胞的培养液中孕酮(P4)含量显著高于含有大卵泡卵母细胞的培养液(分别为276.7和108.2纳克/毫升,p < 0.05)。大卵泡来源的猪卵母细胞具有更高的核成熟和细胞质成熟能力,但差异似乎不是由cAMP介导的。我们的研究结果还表明,小卵泡来源的COCs比大卵泡来源的COCs经历更强烈的黄体化。结果表明,直径大于4.0毫米卵泡中的卵母细胞更适合用于体外研究。

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