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Extracellular pH Dynamically Controls Cell Surface Delivery of Functional TRPV5 Channels

机译：细胞外pH动态控制功能性TRPV5通道的细胞表面递送

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Extracellular pH has long been known to affect the rate and magnitude of ion transport processes among others via regulation of ion channel activity. The Ca2+-selective transient receptor potential vanilloid 5 (TRPV5) channel constitutes the apical entry gate in Ca2+-transporting cells, contributing significantly to the overall Ca2+ balance. Here, we demonstrate that extracellular pH determines the cell surface expression of TRPV5 via a unique mechanism. By a comprehensive approach using total internal reflection fluorescence microscopy, cell surface protein labeling, electrophysiology, 45Ca2+ uptake assays, and functional channel recovery after chemobleaching, this study shows that upon extracellular alkalinization, a pool of TRPV5-containing vesicles is rapidly recruited to the cell surface without collapsing into the plasma membrane. These vesicles contain functional TRPV5 channels since extracellular alkalinization is accompanied by increased TRPV5 activity. Conversely, upon subsequent extracellular acidification, vesicles are retrieved from the plasma membrane, simultaneously resulting in decreased TRPV5 activity. Thus, TRPV5 accesses the extracellular compartment via transient openings of vesicles, suggesting that rapid responses of constitutive active TRP channels to physiological stimuli rely on vesicular “kiss and linger” interactions with the plasma membrane.
机译：长期以来，众所周知，细胞外pH会通过调节离子通道活性来影响离子传输过程的速率和幅度。 Ca 2 + 选择性瞬时受体电位香草酸5（TRPV5）通道构成了Ca 2 + 转运细胞的顶端进入门，对整个Ca 2 + 余额。在这里，我们证明细胞外pH通过独特的机制决定了TRPV5在细胞表面的表达。通过使用全内反射荧光显微镜，细胞表面蛋白标记，电生理， 45 Ca 2 + 吸收测定以及化学漂白后功能通道恢复的综合方法，该研究表明：在细胞外碱化后，含有TRPV5的囊泡会迅速募集到细胞表面，而不会塌陷到质膜中。这些囊泡包含功能性TRPV5通道，因为胞外碱化伴随着TRPV5活性的提高。相反，在随后的细胞外酸化后，从质膜中回收囊泡，同时导致TRPV5活性降低。因此，TRPV5通过短暂的囊泡开口进入细胞外隔室，表明本构性活性TRP通道对生理刺激的快速反应依赖于囊泡与质膜的“亲吻和缠绵”相互作用。

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