Independent regulation of Piezo1 activity by principal and intercalated cells of the collecting duct.
Independent regulation of Piezo1 activity by principal and intercalated cells of the collecting duct.
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DOI：
10.1016/j.jbc.2023.105524
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发表时间：
2024-01
影响因子：
4.8
通讯作者：
Pochynyuk, Oleh
中科院分区：
文献类型：
--
作者：
Pyrshev, Kyrylo;Atamanchuk-Stavniichuk, Anna;Kordysh, Mariya;Zaika, Oleg;Tomilin, Viktor N;Pochynyuk, Oleh
肾脏集合管持续暴露在各种流体流速和渗透梯度下。机械激活的Piezo1通道在集合管中的表达最显著。然而，Piezo1在集合管的功能不同的主细胞和间质细胞(PC和IC)中的地位和调节仍有待确定。我们使用药物Piezo1激活来定量Piezo1介导的新分离的集合管的PC和ICs的[Ca~(2+)]i内流和单通道活性。我们还采用了各种系统治疗来检查它们对PC和IC中Piezo1功能的影响。PIEZO1选择性激动剂Yoda-1或Jedi-2在PC中诱导的Ca~(2+)内流显著高于IC。在膜片钳分析中，我们记录到Yoda-1激活的非选择性通道，其电导为18.6±0.7pS。短期利尿(注射速尿)可刺激PC而非ICs的PIEZO1活性，而抗利尿(限水24小时)可降低PIEZO1活性。然而，高K+饮食对血流的长时间刺激减少了Yoda-1依赖的钙内流，而Piezo1水平没有变化。用NH4Cl补充水诱导代谢性酸中毒可刺激ICs中Piezo1的活性，但不能刺激PC中的Piezo1活性。总体而言，我们的结果显示Piezo1在集合管PC(较多)和ICs(较少)在顶端和基底外侧均有功能表达。我们还发现，流体流量的急剧变化调节Piezo1介导的[Ca~(2+)]i内流，而ICs的通道活动对全身酸碱刺激做出反应。
The renal collecting duct is continuously exposed to a wide spectrum of fluid flow rates and osmotic gradients. Expression of a mechanoactivated Piezo1 channel is the most prominent in the collecting duct. However, the status and regulation of Piezo1 in functionally distinct principal and intercalated cells (PCs and ICs) of the collecting duct remain to be determined. We used pharmacological Piezo1 activation to quantify Piezo1-mediated [Ca2+]i influx and single-channel activity separately in PCs and ICs of freshly isolated collecting ducts with fluorescence imaging and electrophysiological tools. We also employed a variety of systemic treatments to examine their consequences on Piezo1 function in PCs and ICs. Piezo1 selective agonists, Yoda-1 or Jedi-2, induced a significantly greater Ca2+ influx in PCs than in ICs. Using patch clamp analysis, we recorded a Yoda-1-activated nonselective channel with 18.6 ± 0.7 pS conductance on both apical and basolateral membranes. Piezo1 activity in PCs but not ICs was stimulated by short-term diuresis (injections of furosemide) and reduced by antidiuresis (water restriction for 24 h). However, prolonged stimulation of flow by high K+ diet decreased Yoda-1-dependent Ca2+ influx without changes in Piezo1 levels. Water supplementation with NH4Cl to induce metabolic acidosis stimulated Piezo1 activity in ICs but not in PCs. Overall, our results demonstrate functional Piezo1 expression in collecting duct PCs (more) and ICs (less) on both apical and basolateral sides. We also show that acute changes in fluid flow regulate Piezo1-mediated [Ca2+]i influx in PCs, whereas channel activity in ICs responds to systemic acid–base stimuli.