Nicol G D, Schnetkamp P P, Saimi Y, Cragoe E J, Bownds M D
Laboratory of Molecular Biology, University of Wisconsin, Madison 53706.
J Gen Physiol. 1987 Nov;90(5):651-69. doi: 10.1085/jgp.90.5.651.
Vertebrate rod photoreceptors in the dark maintain an inward current across the outer segment membrane. The photoresponse results from a light-induced suppression of this dark current. The light-regulated current is not sensitive to either tetrodotoxin or amiloride, potent blockers of Na+ channels. Here, we report that a derivative of amiloride, 3',4'-dichlorobenzamil (DCPA), completely suppresses the dark current and light response recorded from rod photoreceptors. DCPA also blocks a cyclic GMP-activated current in excised patches of rod plasma membrane and a cGMP-induced Ca++ flux from rod disk membranes. These results are consistent with the notion that the Ca++ flux mechanism in the disk membrane and the light-regulated conductance in the plasma membrane are identical. DCPA also inhibits the Na/Ca exchange mechanism in intact rods, but at a 5-10-fold-higher concentration than is required to block the cGMP-activated flux and current. The blocking action of DCPA in 10 nM Ca++ is different from that in 1 mM Ca++, which suggests either that the conductance state of the light-regulated channel may be modified in high and low concentrations of Ca++, or that there may be two ionic channels in the rod outer segment membrane.
在黑暗中,脊椎动物的视杆光感受器维持着跨外段膜的内向电流。光反应是由光诱导抑制这种暗电流引起的。光调节电流对河豚毒素或氨氯地平(两种有效的Na⁺通道阻滞剂)均不敏感。在此,我们报告氨氯地平的衍生物3',4'-二氯苯甲酰胺(DCPA)完全抑制了视杆光感受器记录到的暗电流和光反应。DCPA还阻断了视杆细胞质膜分离片膜中的环鸟苷酸激活电流以及视杆盘膜中的cGMP诱导的Ca⁺⁺通量。这些结果与盘膜中的Ca⁺⁺通量机制和质膜中的光调节电导相同这一观点一致。DCPA还抑制完整视杆中的Na/Ca交换机制,但所需浓度比阻断cGMP激活通量和电流所需浓度高5至10倍。DCPA在10 nM Ca⁺⁺中的阻断作用与在1 mM Ca⁺⁺中的不同,这表明要么光调节通道的电导状态可能在高浓度和低浓度Ca⁺⁺中发生改变,要么视杆外段膜中可能存在两个离子通道。