Multiple P2X and P2Y receptor subtypes in mouse J774, spleen and peritoneal macrophages
Multiple P2X and P2Y receptor subtypes in mouse J774, spleen and peritoneal macrophages
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DOI：
10.1016/j.bcp.2004.11.012
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发表时间：
2005-02-15
影响因子：
5.8
通讯作者：
Dunn, PM
中科院分区：
文献类型：
--
作者：
Coutinho-Silva, R;Ojcius, DM;Dunn, PM
我们研究了小鼠巨噬细胞和J774细胞系中P2受体的表达和功能，并揭示了比以前认识到的更大的P2受体亚型谱。核苷酸三磷酸腺苷（ATP），二磷酸腺苷，三磷酸尿苷和二磷酸尿苷诱发细胞内钙的增加和钾电流的激活。这些反应对拮抗剂苏拉明、PPADS、MRS 2179和汽巴蓝的敏感性表明存在至少三种功能性P2 Y受体亚型，最可能的是P2 Y（2）、P2 Y（4）和P2 Y（6）。ATP还激活P2 X受体，引起快速激活的阳离子电导。这种反应对拮抗剂苏拉明和汽巴蓝不敏感，被Zn（2+）增强，被酸化抑制，表明P2 X（4）受体参与。在低二价阳离子溶液中，对ATP的反应变得更大，二苯甲酰-ATP变得比ATP更有效，表明存在P2 X（7）受体。免疫荧光、流式细胞术、Western印迹和RT-PCR显示P2 X（4）和P2 X（7）受体在两种巨噬细胞类型中是最突出的，而其他P2 X亚基的表达是可变的，有时很弱或检测不到。这些技术还证实了P2 Y（1）、P2 Y（2）、P2 Y（4）和P2 Y（6）受体的mRNA的存在沿着我们研究的三种亚型（即P2 Y（1）、P2 Y（2）和P2 Y（4））的蛋白质表达。(C)2004年由Elsevier Inc.出版
We investigated P2 receptor expression and function in macrophages from mouse, and in the J774 cell line, and revealed a larger spectrum of P2 receptor subtypes than previously recognised. The nucleotides adenosine triphosphate (ATP), adenosine diphosphate, uridine triphosphate and uridine diphosphate evoked an increase in intracellular calcium and the activation of a potassium current. The sensitivity of these responses to the antagonists suramin, PPADS, MRS 2179 and Cibacron blue suggest the presence of at least three functional P2Y receptor subtypes, most probably P2Y(2), P2Y(4) and P2Y(6). ATP also activated P2X receptors, giving rise to a rapidly activating cation conductance. This response was insensitive to the antagonists suramin and Cibacron blue, was potentiated by Zn(2+) and inhibited by acidification suggesting involvement of P2X(4) receptors. In low divalent cation solution, responses to ATP became larger, and dibenzoyl-ATP became more potent than ATP, indicating the presence of P2X(7) receptors. Immunofluorescence, flow cytometry, Western blots and RT-PCR show that P2X(4) and P2X(7) receptors are the most prominent in both macrophage types, while the expression of the other P2X subunits is variable and sometimes weak or undetectable. These techniques also demonstrated the presence of mRNA for P2Y(1), P2Y(2), P2Y(4) and P2Y(6) receptors along with protein expression for the three subtypes we investigated, namely, P2Y(1), P2Y(2) and P2Y(4). (C) 2004 Published by Elsevier Inc.