A plasma membrane-localized polycystin-1/polycystin-2 complex in endothelial cells elicits vasodilation.
A plasma membrane-localized polycystin-1/polycystin-2 complex in endothelial cells elicits vasodilation.
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DOI：
10.7554/elife.74765
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发表时间：
2022-03-01
期刊：
影响因子：
7.7
通讯作者：
Jaggar JH
中科院分区：
文献类型：
--
作者：
MacKay CE;Floen M;Leo MD;Hasan R;Garrud TAC;Fernández-Peña C;Singh P;Malik KU;Jaggar JH
多囊蛋白-1（PC-1，PKD 1）是由Pkd 1基因表达的受体样蛋白，存在于多种细胞类型中，但其细胞定位、信号传导机制和生理功能尚不清楚。在这里，通过研究他莫昔芬诱导的，内皮细胞（EC）特异性Pkd 1基因敲除（Pkd 1 ecKO）小鼠，我们表明，流量激活PC-1介导的，Ca 2+依赖性阳离子电流的EC。EC特异性PC-1敲除可减弱血流介导的动脉超极化和血管舒张PC-1依赖性血管舒张发生在整个功能性剪切应力范围内，并通过激活内皮一氧化氮合酶（eNOS）和中（IK）和小（SK）电导Ca 2+激活的K+通道。EC特异性PC-1敲除增加全身血压而不改变肾脏解剖结构。PC-1与多囊蛋白-2（PC-2，PKD 2）（TRP多囊蛋白通道）共免疫沉淀，两种蛋白质的簇在EC质膜中以纳米级接近定位。敲除PC-1或PC-2（Pkd 2 ecKO小鼠）可消除EC中PC-1和PC-2的表面簇。PC-1或PC-2的单敲除或PC-1和PC-2的双敲除（Pkd 1/Pkd 2 ecKO小鼠）类似地减弱血流介导的血管舒张。流量刺激EC中的非选择性阳离子电流，这些阳离子电流同样受到PC-1或PC-2敲除或PC-1或PC-2中存在的C-末端卷曲螺旋结构域对应的干扰肽的抑制。总之，我们发现PC-1通过与PC-2在EC中形成相互依赖的信号复合物来调节动脉收缩力。血流刺激EC质膜中的PC-1/PC-2簇，导致eNOS、IK通道和SK通道激活、血管舒张和血压降低。
Polycystin-1 (PC-1, PKD1), a receptor-like protein expressed by the Pkd1 gene, is present in a wide variety of cell types, but its cellular location, signaling mechanisms, and physiological functions are poorly understood. Here, by studying tamoxifen-inducible, endothelial cell (EC)-specific Pkd1 knockout (Pkd1 ecKO) mice, we show that flow activates PC-1-mediated, Ca2+-dependent cation currents in ECs. EC-specific PC-1 knockout attenuates flow-mediated arterial hyperpolarization and vasodilation. PC-1-dependent vasodilation occurs over the entire functional shear stress range and via the activation of endothelial nitric oxide synthase (eNOS) and intermediate (IK)- and small (SK)-conductance Ca2+-activated K+ channels. EC-specific PC-1 knockout increases systemic blood pressure without altering kidney anatomy. PC-1 coimmunoprecipitates with polycystin-2 (PC-2, PKD2), a TRP polycystin channel, and clusters of both proteins locate in nanoscale proximity in the EC plasma membrane. Knockout of either PC-1 or PC-2 (Pkd2 ecKO mice) abolishes surface clusters of both PC-1 and PC-2 in ECs. Single knockout of PC-1 or PC-2 or double knockout of PC-1 and PC-2 (Pkd1/Pkd2 ecKO mice) similarly attenuates flow-mediated vasodilation. Flow stimulates nonselective cation currents in ECs that are similarly inhibited by either PC-1 or PC-2 knockout or by interference peptides corresponding to the C-terminus coiled-coil domains present in PC-1 or PC-2. In summary, we show that PC-1 regulates arterial contractility through the formation of an interdependent signaling complex with PC-2 in ECs. Flow stimulates PC-1/PC-2 clusters in the EC plasma membrane, leading to eNOS, IK channel, and SK channel activation, vasodilation, and a reduction in blood pressure.