Knopp A, Rüppel H
Max Volmer Institut für Biophysikalische and Physikalische Chemie, Technische Universität Berlin, Germany.
J Gen Physiol. 1996 May;107(5):577-95. doi: 10.1085/jgp.107.5.577.
By use of microelectrodes, changes in the receptor current and the Ca2+ concentration were measured in the rod layer of the rat retina after stimulation by flashes or steady light. Thereby light induced Ca2+ sources, and sinks along a rod were determined in dependence of time. Thus, the Ca2+ fluxes across the plasma membrane of a mammalian rod could be studied in detail. By light stimulation, Ca2+ sources are evoked along the outer segment only. Immediately after a saturating flash, a maximum of Ca2+ efflux is observed which decays exponentially with tau = 0.3 s at 37 degrees C (4.2 s at 23 degrees C). During regeneration of the dark current, the outer segment acts as a Ca2+ sink, indicating a restoration of the Ca(2+)-depleted outer segment. These findings agree with earlier reports on amphibian rods. Further experiments showed that the peak Ca2+ efflux and tau are temperature dependent. The peak amplitude also depends on the external Ca2+ concentration. In contrast to the reports on amphibian rods, only a part of the Ca2+ ions extruded from the outer segment is directly restored. Surprisingly, during steady light the Ca2+ efflux approaches a permanent residual value. Therefore, in course of a photoresponse, Ca2+ must be liberated irreversibly from internal Ca2+ stores. There is certain evidence that the inner segment acts as a Ca2+ store. Our results show that the Ca2+ fraction of the ions carrying the dark current is proportional to the extracellular Ca2+ concentration. This indicates that the Ca2+ permeability of the plasma membrane of the rod outer segment is independent of the Ca2+ concentration.
通过使用微电极,在大鼠视网膜的视杆层中,测量了闪光或持续光照刺激后受体电流和Ca2+浓度的变化。据此确定了视杆上光诱导的Ca2+来源和汇随时间的变化。因此,可以详细研究哺乳动物视杆细胞膜上的Ca2+通量。通过光刺激,仅在外段诱发出Ca2+来源。在饱和闪光后立即观察到最大的Ca2+外流,在37℃时其以时间常数τ = 0.3秒呈指数衰减(在23℃时为4.2秒)。在暗电流再生期间,外段充当Ca2+汇,表明Ca2+耗尽的外段得到恢复。这些发现与早期关于两栖动物视杆的报道一致。进一步的实验表明,Ca2+外流峰值和时间常数与温度有关。峰值幅度也取决于外部Ca2+浓度。与关于两栖动物视杆的报道不同,从外段挤出的Ca2+离子只有一部分被直接恢复。令人惊讶的是,在持续光照期间,Ca2+外流接近一个永久的残余值。因此,在光反应过程中,Ca2+必须从内部Ca2+储存库中不可逆地释放出来。有一定证据表明内段充当Ca2+储存库。我们的结果表明,携带暗电流的离子中的Ca2+部分与细胞外Ca2+浓度成正比。这表明视杆外段质膜的Ca2+通透性与Ca2+浓度无关。