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卵母细胞切除术不影响猪卵丘细胞透明质酸的合成:在无血清培养基中用胰岛素样生长因子-I处理后透明质酸的保留情况

Oocytectomy does not influence synthesis of hyaluronic acid by pig cumulus cells: retention of hyaluronic acid after insulin-like growth factor-I treatment in serum-free medium.

作者信息

Nagyová E, Procházka R, Vanderhyden B C

机构信息

Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov, Czech Republic.

出版信息

Biol Reprod. 1999 Sep;61(3):569-74. doi: 10.1095/biolreprod61.3.569.

Abstract

Mouse oocytes secrete a factor that enables cumulus cells to undergo expansion in response to FSH (1 microg/ml), whereas expansion of the porcine cumulus oophorus has been shown to be independent of the oocyte. The aim of this study was to assess FSH-induced synthesis of hyaluronic acid (HA) by porcine cumulus cells before and after oocytectomy. In addition, we studied the effect of insulin-like growth factor-I (IGF-I) on the ability of cumulus cells to synthesize and retain HA in response to FSH in serum-free medium. Porcine oocyte-cumulus complexes and complexes from which the oocytes had been removed by oocytectomy were cultured for 24 h in the presence of 2.5 microCi of D-[6-(3)H]glucosamine hydrochloride, fetal calf serum (FCS, 5%), and FSH. After 24 h, incorporation of [(3)H]glucosamine into HA was measured either in complexes alone (retained HA) or in medium plus complexes (total HA). Specificity of incorporation of radioactivity into HA was confirmed by the sensitivity to highly specific Streptomyces hyaluronidase. Our results suggest that 1) the synthesis of HA by pig cumulus cells in vitro is stimulated by FSH and that oocytectomy does not change this synthesis; 2) oocytes do not influence retention of HA within the complex; 3) FSH-induced synthesis of HA by cumulus cells is decreased in medium with polyvinylpyrrolidone (PVP)-supplemented (total and retained HA) compared to FCS-supplemented medium; 4) IGF-I enabled cumulus cells to synthesize HA in response to FSH in PVP-supplemented medium in a manner similar to that observed when serum is present in the medium.

摘要

小鼠卵母细胞分泌一种因子,该因子能使卵丘细胞在促卵泡激素(FSH,1微克/毫升)作用下发生扩张,而猪卵丘-卵母细胞复合体的扩张已被证明与卵母细胞无关。本研究的目的是评估卵母细胞切除前后,FSH诱导猪卵丘细胞合成透明质酸(HA)的情况。此外,我们还研究了胰岛素样生长因子-I(IGF-I)对卵丘细胞在无血清培养基中响应FSH合成和保留HA能力的影响。将猪卵母细胞-卵丘复合体以及通过卵母细胞切除术去除卵母细胞后的复合体,在含有2.5微居里D-[6-(3)H]盐酸葡萄糖胺、胎牛血清(FCS,5%)和FSH的条件下培养24小时。24小时后,测量[(3)H]葡萄糖胺掺入HA的情况,可单独在复合体中测量(保留的HA),也可在培养基加复合体中测量(总HA)。通过对高特异性透明质酸酶的敏感性,证实了放射性掺入HA的特异性。我们的结果表明:1)FSH刺激猪卵丘细胞在体外合成HA,卵母细胞切除不改变这种合成;2)卵母细胞不影响HA在复合体内的保留;3)与补充FCS的培养基相比,在补充聚乙烯吡咯烷酮(PVP)的培养基中,FSH诱导卵丘细胞合成HA减少(总HA和保留的HA);4)IGF-I使卵丘细胞在补充PVP的培养基中能够响应FSH合成HA,其方式与培养基中存在血清时观察到的相似。

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