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碱性成纤维细胞生长因子(bFGF)受体在大鼠卵巢黄体细胞中随黄体年龄增加而减少:黄体细胞中bFGF受体与bFGF的共定位

Basic fibroblast growth factor (bFGF) receptors decrease with luteal age in rat ovarian luteal cells: colocalization of bFGF receptors and bFGF in luteal cells.

作者信息

Asakai R, Tamura K, Eishi Y, Iwamoto M, Kato Y, Okamoto R

机构信息

Division of Functional Disorder Research, School of Medicine, Tokyo Medical and Dental University, Japan.

出版信息

Endocrinology. 1993 Sep;133(3):1074-84. doi: 10.1210/endo.133.3.7689947.

Abstract

Ovarian growth factors have been implicated in the development and differentiation of corpus luteum. We have characterized both high and low affinity receptors for basic fibroblast growth factor (bFGF) in luteal cells and tissue throughout the life span of corpus luteum using gonadotropin-treated rat luteinized ovaries. Additionally, we determined bFGF location in luteal tissue. High affinity (Kd, approximately 0.2 nM) and low capacity (approximately 500-6000 sites/cell, depending on luteal ages) [125I] bFGF-binding sites (mol wt, 140 kilodaltons, determined by affinity labeling) were found on luteal cells. [125I]bFGF binding to luteal cells and corpus luteum membranes progressively decreased in binding capacity without affecting binding affinity as the age of corpus luteum advanced. bFGF receptor (flg) mRNA in luteinized ovaries decreased with the luteal age similar to the [125I]bFGF binding. In contrast, low affinity binding sites, identified as heparan sulfate proteoglycans, which were immunohistochemically visualized around luteal cells, did not appear to change with luteal age. The signals for heparan sulfate proteoglycans that were found only in granulosa, but not theca-interstitial, cells of follicles became intense during folliculogenesis. The functionality of the bFGF receptors was shown by tyrosine phosphorylation of 16.5- and 18-kilodalton proteins in luteal cells with the antiphosphotyrosine antibody. Immunohistochemistry localized bFGF to steroidogenic luteal cells, and immunoblotting displayed larger molecular forms of bFGF in luteal cells. These results suggest that the bFGF receptors may be associated with the development and differentiation of corpus luteum in an autocrine manner.

摘要

卵巢生长因子与黄体的发育和分化有关。我们利用促性腺激素处理的大鼠黄体化卵巢,在黄体整个生命周期的黄体细胞和组织中,对碱性成纤维细胞生长因子(bFGF)的高亲和力和低亲和力受体进行了表征。此外,我们还确定了bFGF在黄体组织中的位置。在黄体细胞上发现了高亲和力(Kd,约0.2 nM)和低容量(约500 - 6000个位点/细胞,取决于黄体年龄)的[125I]bFGF结合位点(通过亲和标记测定分子量为140千道尔顿)。随着黄体年龄的增加,[125I]bFGF与黄体细胞和黄体细胞膜的结合能力逐渐下降,但不影响结合亲和力。黄体化卵巢中的bFGF受体(flg)mRNA随着黄体年龄的增加而减少,与[125I]bFGF结合情况相似。相比之下,被鉴定为硫酸乙酰肝素蛋白聚糖的低亲和力结合位点,通过免疫组织化学在黄体细胞周围可见,其数量似乎不随黄体年龄变化。仅在卵泡的颗粒细胞而非卵泡膜间质细胞中发现的硫酸乙酰肝素蛋白聚糖信号,在卵泡发生过程中增强。用抗磷酸酪氨酸抗体显示,黄体细胞中16.5和18千道尔顿蛋白的酪氨酸磷酸化表明了bFGF受体的功能。免疫组织化学将bFGF定位于类固醇生成黄体细胞,免疫印迹显示黄体细胞中有更大分子形式的bFGF。这些结果表明,bFGF受体可能以自分泌方式参与黄体的发育和分化。

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