Young J D, Unkeless J C, Kaback H R, Cohn Z A
Proc Natl Acad Sci U S A. 1983 Mar;80(5):1357-61. doi: 10.1073/pnas.80.5.1357.
We have studied the effects of specific ligands of the receptor for the IgG Fc fragment (FcR) on the membrane potential (delta psi) of the macrophage cell line J774 by the [3H]tetraphenylphosphonium ion equilibration technique. We observe a membrane depolarization with binding of FcR ligands that is dependent on the degree of receptor crosslinking. Binding of the FcR by monovalent ligands is not sufficient to induce a significant drop in delta psi, but a sustained depolarization lasting approximately equal to 20 min occurs with insoluble multivalent ligands. This FcR-mediated depolarization can be inhibited by substitution of Na+ from the cell incubation medium with monovalent choline cation, indicating that depolarization is due to Na+ influx into the cell. The extracellular Ca2+ does not play a significant role in membrane depolarization. The depolarization response is not triggered by monoclonal antibodies directed against three other major macrophage surface antigens. The cell depolarization mediated by FcR ligands is followed by a prolonged hyperpolarization that can be partially blocked by ouabain and quinine, indicating that the hyperpolarization response is a result of a combination of a Na+,K+-ATPase activity and a Ca2+-activated K+ conductance. These data support our hypothesis that the mouse macrophage IgG FcR is a ligand-dependent ion channel.
我们采用[3H]四苯基鏻离子平衡技术,研究了IgG Fc片段受体(FcR)的特异性配体对巨噬细胞系J774膜电位(δψ)的影响。我们观察到FcR配体结合会导致膜去极化,这取决于受体交联程度。单价配体与FcR的结合不足以引起δψ的显著下降,但不溶性多价配体可导致持续约20分钟的去极化。这种FcR介导的去极化可通过用单价胆碱阳离子替代细胞培养液中的Na+来抑制,表明去极化是由于Na+流入细胞所致。细胞外Ca2+在膜去极化中不起重要作用。针对其他三种主要巨噬细胞表面抗原的单克隆抗体不会引发去极化反应。FcR配体介导的细胞去极化之后是长时间的超极化,哇巴因和奎宁可部分阻断这种超极化,表明超极化反应是Na+,K+-ATP酶活性和Ca2+激活的K+电导共同作用的结果。这些数据支持了我们的假设,即小鼠巨噬细胞IgG FcR是一种配体依赖性离子通道。