Sakura H, Ashcroft F M
University Laboratory of Physiology, Oxford, UK.
Diabetologia. 1997 May;40(5):528-32. doi: 10.1007/s001250050711.
Insulin secretion is stimulated by glucose, hormones and neurotransmitters. Both activation of a non-selective cation current and activation of a Ca2+ current in response to depletion of intracellular Ca2+ stores have been suggested to play a role in this stimulation. The properties of these currents resemble those reported for the Drosophila genes trp and trpl. Using the reverse transcription polymerase chain reaction and Northern blot analysis we found that of the six mammalian trp-related genes (trp1-6), only trp1 was expressed at high levels in the mouse insulinoma cell line MIN6. We cloned the murine homologue of human trp1 from MIN6 cells and identified four variants (alpha, beta, gamma and delta), generated by alternative splicing near the N-terminus of the protein. In vitro translation showed that only the alpha and beta splice variants are efficiently expressed. The beta variant is the dominant form in MIN6 cells (and probably in mouse pancreatic islets), whereas the alpha variant is the major type in the mouse brain. The beta variant showed 99% identity to the human homologue at the amino acid level.
胰岛素分泌受葡萄糖、激素和神经递质的刺激。有研究表明,响应细胞内钙库耗竭时,非选择性阳离子电流的激活以及钙电流的激活在这种刺激过程中发挥作用。这些电流的特性与果蝇基因trp和trpl所报道的特性相似。通过逆转录聚合酶链反应和Northern印迹分析,我们发现,在六个哺乳动物trp相关基因(trp1 - 6)中,只有trp1在小鼠胰岛素瘤细胞系MIN6中高水平表达。我们从MIN6细胞中克隆了人trp1的小鼠同源物,并鉴定出四种变体(α、β、γ和δ),它们是由蛋白质N端附近的可变剪接产生的。体外翻译表明,只有α和β剪接变体能够有效表达。β变体是MIN6细胞(可能还有小鼠胰岛)中的主要形式,而α变体是小鼠脑中的主要类型。β变体在氨基酸水平上与人同源物有99%的同一性。