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Increasing the expression of calcium-permeable TRPC3 and TRPC7 channels enhances constitutive secretion

机译:增加钙可渗透的TRPC3和TRPC7通道的表达可增强本构分泌

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摘要

The hTRPC [human TRPC (canonical transient receptor potential)] family of non-selective cation channels is proposed to mediate calcium influx across the plasma membrane via PLC (phospholipase C)-coupled receptors. Heterologously expressed hTRPC3 and hTRPC7 have been localized at the cell surface; however, a large intracellular component has also been noted but not characterized. In the present study, we have investigated the intracellular pool in COS-7 cells and have shown co-localization with markers for both the TGN (trans-Golgi network) and the cis-Golgi cisternae by immunofluorescence microscopy. Addition of BFA (Brefeldin A) to cells expressing hTRPC3 or hTRPC7 resulted in the redistribution of the Golgi component to the endoplasmic reticulum, indicating that this pool is present in both the Golgi stack and the TGN. Expression of either TRPC3 or TRPC7, but not TRPC1 or the cell surface marker CD8, resulted in a 2–4-fold increase in secreted alkaline phosphatase in the extracellular medium. Based on these results, we propose that an additional function of these members of the hTRPC family may be to enhance secretion either by affecting transport through the Golgi stack or by increasing fusion at the plasma membrane.
机译:提出了非选择性阳离子通道的hTRPC [人类TRPC(规范瞬时受体电位)]家族,可通过PLC(磷脂酶C)偶联的受体介导跨质膜的钙流入。异源表达的hTRPC3和hTRPC7已定位在细胞表面。然而,也已经注意到了大的细胞内成分,但是没有被表征。在本研究中,我们研究了COS-7细胞的细胞内池,并通过免疫荧光显微镜显示了与TGN(反式高尔基体网络)和顺式高尔基体池的标志物共定位。在表达hTRPC3或hTRPC7的细胞中添加BFA(布雷菲德菌素A)导致高尔基体成分重新分布到内质网,表明该池同时存在于高尔基体和TGN中。 TRPC3或TRPC7的表达而不是TRPC1或细胞表面标记CD8的表达导致细胞外培养基中分泌的碱性磷酸酶增加2-4倍。基于这些结果,我们建议hTRPC家族的这些成员的另一个功能可能是通过影响通过高尔基体的运输或通过增加质膜的融合来增强分泌。

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