Harbeck M C, Louie D C, Howland J, Wolf B A, Rothenberg P L
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, 19104, USA.
Diabetes. 1996 Jun;45(6):711-7. doi: 10.2337/diab.45.6.711.
The expression of insulin receptor mRNA was examined in rat pancreatic islet cells by single-cell reverse transcriptase (RT)-polymerase chain reaction (PCR). Single cells from disaggregated islets were individually isolated in a microcapillary pipet, and the beta-cells were identified by amplification of the mRNA for insulin. We found that in single beta-cells, the mRNA for the insulin receptor was also expressed. The fraction of single islet cells expressing both insulin receptor and insulin mRNAs corresponds closely to the fraction of beta-cells in the disaggregated islet cell preparation. These results indicate that normal beta-cells have the potential to express authentic insulin receptors. Immunohistochemical analysis was insufficiently sensitive for assaying insulin receptor protein; however, insulin receptor substrate 1 (IRS-1) was readily immunolocalized in islet beta-cells. Since IRS-1 links several cell surface receptors, including those for insulin and IGF-I, to distal signal transduction pathways, our observations indicate that hormonal regulation of islet beta-cells potentially involves the same signal transduction pathway that mediates insulin and growth factor signaling in peripheral insulin target tissue cell types.
采用单细胞逆转录酶(RT)-聚合酶链反应(PCR)技术检测大鼠胰岛细胞中胰岛素受体mRNA的表达。将分散的胰岛中的单细胞逐个分离到微量毛细管移液管中,通过胰岛素mRNA的扩增来鉴定β细胞。我们发现,在单个β细胞中,胰岛素受体mRNA也有表达。同时表达胰岛素受体和胰岛素mRNA的单个胰岛细胞的比例与分散的胰岛细胞制备物中β细胞的比例密切相关。这些结果表明,正常β细胞具有表达真实胰岛素受体的潜力。免疫组织化学分析检测胰岛素受体蛋白的敏感性不足;然而,胰岛素受体底物1(IRS-1)很容易在胰岛β细胞中进行免疫定位。由于IRS-1将包括胰岛素和IGF-I受体在内的几种细胞表面受体与远端信号转导途径联系起来,我们的观察结果表明,胰岛β细胞的激素调节可能涉及与外周胰岛素靶组织细胞类型中介导胰岛素和生长因子信号传导相同的信号转导途径。