Gorr S U
Endocrine Cell Biology Laboratory, Department of Biological and Biophysical Sciences, University of Louisville Health Sciences Center, Louisville, Kentucky 40292, USA.
J Biol Chem. 1996 Feb 16;271(7):3575-80. doi: 10.1074/jbc.271.7.3575.
The rat pituitary cell line GH4C1 secretes granins (chromogranin B and secretogranin II) and prolactin by the regulated secretory pathway. The intracellular storage of prolactin is preferentially induced by hormone treatment with estradiol, insulin, and epidermal growth factor. The goal of this study was to determine the effect of hormone treatment on storage of granins and constitutive secretory markers. The granins were efficiently stored in both hormone-treated and -untreated cells (17% of total secreted in 4 h). Secreted alkaline phosphatase (SEAP), a truncated membrane protein that would not be expected to enter secretory granules, and glycosaminoglycan, a marker for the constitutive secretory pathway, exhibited 70 80% secretion under both conditions. In comparison, the relative prolactin secretion was 31 and 68% from hormone-treated and -untreated cells, respectively. Phorbol ester and KCl stimulated prolactin secretion 2.3-fold from untreated cells and 5. 5-fold from hormone-treated cells. In contrast, SEAP secretion was stimulated 1.5-fold from both treated and untreated cells, consistent with secretion by the constitutive secretory pathway. Stimulated secretion of granins was detected from both hormone-treated and -untreated cells. These results suggest that granin and prolactin storage are differentially regulated and that the constitutive secretory pathway is not affected by hormone treatment.
大鼠垂体细胞系GH4C1通过调节性分泌途径分泌颗粒蛋白(嗜铬粒蛋白B和分泌粒蛋白II)和催乳素。用雌二醇、胰岛素和表皮生长因子进行激素处理可优先诱导催乳素的细胞内储存。本研究的目的是确定激素处理对颗粒蛋白储存和组成型分泌标志物的影响。颗粒蛋白在激素处理和未处理的细胞中均能有效储存(4小时内占总分泌量的17%)。分泌性碱性磷酸酶(SEAP)是一种预计不会进入分泌颗粒的截短膜蛋白,以及组成型分泌途径的标志物糖胺聚糖,在两种条件下的分泌率均为70%至80%。相比之下,激素处理和未处理细胞的催乳素相对分泌率分别为31%和68%。佛波酯和氯化钾刺激未处理细胞的催乳素分泌增加2.3倍,刺激激素处理细胞的催乳素分泌增加5.5倍。相比之下,处理和未处理细胞的SEAP分泌均增加1.5倍,这与组成型分泌途径的分泌情况一致。在激素处理和未处理的细胞中均检测到颗粒蛋白的刺激分泌。这些结果表明,颗粒蛋白和催乳素的储存受到不同的调节,并且组成型分泌途径不受激素处理的影响。