马卵巢类固醇急性调节蛋白的检测。

Elaine D. Watson, S. R. Thomson, A. Howie
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引用次数: 15

摘要

一种类固醇急性调节蛋白(StAR)已经在一些物种中被鉴定为可能是类固醇形成中重要的限速步骤。这种蛋白质被认为负责将胆固醇从线粒体外膜运输到线粒体内膜。已知马绒毛膜促性腺激素(eCG)刺激早孕母马黄体的甾体生成,并且eCG也上调牛卵巢中的StAR mRNA。本研究用免疫染色法检测了妊娠期和早孕母马卵巢组织中StAR蛋白的分布。进行Western blot分析,然后进行荧光成像,以确定妊娠期eCG分泌的开始是否与StAR蛋白表达增加有关。StAR的免疫染色仅限于生长卵泡内膜和排卵前卵泡,但用hCG处理24 h后,一些颗粒细胞被阳性染色。阳性染色仅限于马黄体的大黄体细胞。妊娠母马心电图分泌开始前后免疫染色分布无差异,但妊娠第40天和第41天与未妊娠母马和妊娠第20-30天的母马相比,黄体中StAR含量增加。
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Detection of steroidogenic acute regulatory protein in equine ovaries.
A steroidogenic acute regulatory (StAR) protein has been identified in several species as a probable important rate-limiting step in steroidogenesis. This protein is believed to be responsible for transporting cholesterol from the outer to the inner mitochondrial membrane. It is known that equine chorionic gonadotrophin (eCG) stimulates steroidogenesis in the corpora lutea of early pregnant mares and that eCG also upregulates StAR mRNA in bovine ovaries. In the present study, ovarian tissue from cyclic and early pregnant mares was immunostained to detect the distribution of the StAR protein. Western blot analysis was performed, followed by phosphor imaging to establish whether the onset of eCG secretion in pregnancy was associated with increased expression of the StAR protein. Immunostaining for StAR was confined to the theca interna of growing and preovulatory follicles, but 24 h after treatment with hCG, some granulosa cells were positively stained. Positive staining was confined to the large luteal cells of the equine corpus luteum. There was no difference in the distribution of immunostaining before or after onset of eCG secretion in pregnant mares, but increased amounts of StAR were detected in corpora lutea from mares at day 40 or day 41 of pregnancy compared with non-pregnant mares and mares at days 20-30 of pregnancy.
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