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卵母细胞分泌的因子可提高小卵泡卵母细胞体外成熟培养体系中原代卵母细胞的胚胎发育能力。

Oocyte secreted factors improve embryo developmental competence of COCs from small follicles in prepubertal goats.

机构信息

Departament de Ciència Animal i dels Aliments, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.

出版信息

Theriogenology. 2010 Oct 1;74(6):1050-9. doi: 10.1016/j.theriogenology.2010.04.035. Epub 2010 Jun 9.

Abstract

Oocytes secrete soluble paracrine factors called Oocyte Secreted Factors (OSFs) which regulate the cumulus cell phenotype. Follicle populations in ovaries from prepubertal females have smaller diameters than their adult counterparts. Oocytes from small follicles are less competent than those from large follicles. The aim of this study was to investigate, in prepubertal goats, the effect of OSFs secreted by denuded oocytes (DOs) from small (<3 mm) or large (>or=3 mm) follicles during IVM on embryo development and the blastocyst quality of cumulus-oocyte complexes (COCs) from small follicles and to determine if GDF9 participates in this process. Treatment groups were: (A) COCs non selected by their follicle size (control group); (B) cumulus oocytes complexes from small follicles (SFCOCs), (C) cumulus oocytes complexes from small follicles co-cultured with denuded oocytes from small follicles (SFCOCs + SFDOs), and (D) cumulus oocytes complexes from small follicles co-cultured with denuded oocytes from large follicles (SFCOCs + LFDOs). The effect of the addition of kinase inhibitor SB-431542, which antagonizes GDF9, was tested in A, C, and D treatment groups. Co-cultured SFCOCs with SFDOs or LFDOs significantly augmented the blastocyst rate in comparison to SFCOCs alone (15.77%, 17.39% vs. 10.31%, respectively). Blastocysts from SFCOCs + LFDOs group showed higher rates of tetraploid nuclei than blastocysts from SFCOCs and the control group (14.43% vs. 5.45% and 5.24%, respectively; P < 0.05). However, we did not observe differences in the hatching rate, mean cell number or embryo cryotolerance (P > 0.05) between the four treatment groups. The addition of SB-431542 during IVM did not have any effect on blastocyst rate (P > 0.05). In conclusion, in prepubertal goats, COCs with a low embryo developmental competence as a consequence of follicle size can be improved by coculturing them with denuded oocytes from both small and large follicles. GDF9 does not seem play a role in this improvement.

摘要

卵母细胞分泌可溶性旁分泌因子,称为卵母细胞分泌因子 (OSFs),它们调节卵丘细胞表型。青春期前女性卵巢中的卵泡群直径小于成年女性。小卵泡中的卵母细胞不如大卵泡中的卵母细胞有能力。本研究旨在探讨青春期前山羊中,在 IVM 期间,来自小 (<3mm) 或大 (>或=3mm) 卵泡的去卵丘卵母细胞 (DOs) 分泌的 OSFs 对小卵泡卵母细胞的胚胎发育和囊胚质量的影响,并确定 GDF9 是否参与该过程。处理组为:(A) 未按卵泡大小选择的卵母细胞复合物 (对照组);(B) 小卵泡卵母细胞复合物 (SFCOCs);(C) 与小卵泡去卵丘卵母细胞共培养的小卵泡卵母细胞复合物 (SFCOCs+SFDOs);(D) 与大卵泡去卵丘卵母细胞共培养的小卵泡卵母细胞复合物 (SFCOCs+LFDOs)。在处理组 A、C 和 D 中测试了添加激酶抑制剂 SB-431542 的效果,该抑制剂拮抗 GDF9。与单独的 SFCOCs 相比,共培养的 SFCOCs 与 SFDOs 或 LFDOs 显著提高了囊胚率(分别为 15.77%、17.39% 和 10.31%)。来自 SFCOCs+LFDOs 组的囊胚核四倍体率高于 SFCOCs 和对照组的囊胚核(分别为 14.43%、5.45% 和 5.24%;P<0.05)。然而,在四个处理组之间,孵化率、平均细胞数或胚胎冷冻耐受性没有差异(P>0.05)。在 IVM 期间添加 SB-431542 对囊胚率没有影响(P>0.05)。综上所述,在青春期前的山羊中,由于卵泡大小导致胚胎发育能力低的卵母细胞复合物可以通过与小卵泡和大卵泡的去卵丘卵母细胞共培养来提高。GDF9 在这种改善中似乎没有发挥作用。

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