P2X4 and P2X6 receptors associate with VE-cadherin in human endothelial cells
P2X4 and P2X6 receptors associate with VE-cadherin in human endothelial cells
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DOI：
10.1007/s00018-002-8474-y
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发表时间：
2002-05-01
影响因子：
8
通讯作者：
Burnstock, G
中科院分区：
文献类型：
--
作者：
Glass, R;Loesch, A;Burnstock, G
我们研究了P2X(4)和P2X(6)受体在人脐静脉内皮细胞（HUVECs）上的表达，发现质膜上的两种P2X受体亚型在很大程度上局限于细胞-细胞接触区域。共聚焦和电镜水平的共标记实验显示，P2X(4)和P2X(6)受体与细胞粘附分子VE-cadherin强烈共定位。细胞连接处质膜上的P2X(4)和P2X(6)受体在细胞外[Ca2+]减少后迅速（在5分钟内）特异性内化。微丝、微管的破坏和整合素介导的粘附或ATP刺激P2受体不会改变HUVEC质膜上P2X(4)和P2X(6)受体的表达。膜质P2X(4)和P2X(6)受体抵抗Triton-X 100提取，而细胞质P2X受体可溶于Triton-X 100。P2X(4)受体可与VE-cadherin共免疫沉淀，而P2X(6)受体不能与VE-cadherin共免疫沉淀，反之亦然。我们得出结论，P2X(4)和P2X(6)受体在HUVEC粘附连接处与VE-cadherin相关。
We investigated the expression of P2X(4) and P2X(6) receptors on human umbilical vein endothelial cells (HUVECs) and found that both P2X receptor subtypes on plasma membranes are largely restricted to areas of cell-cell contact. Co-labelling experiments at the confocal and electron microscopy levels revealed that P2X(4) and P2X(6) receptors are strongly co-localised with the cell adhesion molecule VE-cadherin. The P2X(4) and P2X(6) receptors on plasma membranes at cellular junctions are rapidly (within 5 min) internalised specifically after decreasing extracellular [Ca2+]. Disruption of microfilaments, microtubules and integrin-mediated adhesion or stimulation of P2 receptors with ATP did not alter P2X(4) and P2X(6) receptor expression on HUVEC plasma membranes. Membraneous P2X(4) and P2X(6) receptors resisted extraction with Triton-X 100, whereas cytoplasmic P2X receptors were Triton-X 100 soluble. P2X(4) receptors, but not P2X(6) receptors, could be co-immunoprecipitated with VE-cadherin and vice versa. We conclude that P2X(4) and P2X(6) receptors are associated with VE-cadherin at HUVEC adherens junctions.