P2X currents in peritoneal macrophages of wild type and P2X4-/- mice
P2X currents in peritoneal macrophages of wild type and P2X4-/- mice
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DOI：
10.1016/j.imlet.2007.07.015
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发表时间：
2007-11-15
影响因子：
4.4
通讯作者：
Meert, Theo
中科院分区：
文献类型：
--
作者：
Brone, Bert;Moechars, Diederik;Meert, Theo
本研究采用全细胞膜片钳技术研究了野生型（WT）和P2X(4)敲除型(P2X（4)(-/-)）小鼠腹膜巨噬细胞中atp诱导的（P2X）电流，目的是：(1)研究天然巨噬细胞的P2X电流，(2)研究P2X(4)样电流在野生型（WT）和P2X(4)（-/-）小鼠腹膜巨噬细胞中的表达。在分离的巨噬细胞中观察到三种类型的电流：(1)在大约10%的WT和P2X4-/-巨噬细胞中，低浓度(0。1-1 μ M)的ATP；(2) 85%的野生型和100%的P2X(4)（-/-）巨噬细胞在高浓度ATP （10 mM）激活下表现出非脱敏的P2X(7)样电流。使用特定阻断剂A-740003确认P2X(7)电流的身份；(3) 88.6%的WT巨噬细胞（P2X4-/-）在中等浓度（3-300 μ M）的ATP作用下表现出小的P2X4样电流，缓慢脱敏。多次观察表明，WT巨噬细胞的缓慢脱敏电流为P2X4。在P2X4-/-小鼠中，5.3 μ M ATP的EC50值与预期一致，3-300 μ M ATP诱导的电流在P2X4-/-小鼠中缺失。应用3 μ M ivermectin（一种P2X(4)选择性调制剂）后，该电流的振幅增加，并且在WT细胞中抑制脱敏。此外，该电流被10 μ M Zn2+促进，但被Cu2+抑制（与P2X(2)相反）。我们得出结论，P2X4和P2X7电流在WT小鼠募集的腹膜巨噬细胞中功能性表达，而P2X(4)（-/-）小鼠中不存在P2X4样电流。(C) 2007 Elsevier B.V.版权所有
In this study the ATP-induced (P2X) currents in isolated peritoneal macrophages of wild type (WT) and P2X(4) knockout (P2X(4)(-/-)) mice were studied by means of whole-cell patch clamp in order to (1) survey the P2X currents of native macrophages and (2) to investigate the expression of P2X(4)-like currents in the WT versus P2X(4)(-/-) mice. Three types of currents were observed in the isolated macrophages: (1) in similar to 10% of both WT and P2X4-/- macrophages a fast activating and inactivating P2X1-like current was recorded with low concentrations (0. 1-1 mu M) of ATP; (2) 85% of wild type and 100% of P2X(4) (-/-) macrophages exhibited a non-desensitizing P2X(7)-like current activated at high concentrations of ATP (10 mM). The identity of the P2X(7) current was confirmed using the specific blocker A-740003; (3) 88.6% of the WT but none of the P2X4-/- macrophages showed a small P2X4-like current that desensitized slowly upon ATP application at intermediate concentrations (3-300 mu M). Several observations indicated that the slowly desensitizing current in WT macrophages was P2X4. The EC50 value of 5.3 mu M ATP was as expected for P2X4 and the current induced by 3-300 mu M ATP was absent in P2X4-/- mice. Upon application of 3 mu M ivermectin, a P2X(4)-selective modulator, the amplitude of this current was increased and the desensitization was inhibited in WT cells. In addition, this current was facilitated by 10 mu M Zn2+ but inhibited by Cu2+ (in contrast to P2X(2)). We conclude that the P2X4 and P2X7 currents are functionally expressed in recruited peritoneal macrophages of WT mice and that the P2X4-like current is absent in P2X(4)(-/-) mice. (C) 2007 Elsevier B.V. All rights reserved.