Harsanyi K, Mangel S C
Department of Ophthalmology, University of Alabama School of Medicine, Birmingham 35294.
Vis Neurosci. 1993 Jan-Feb;10(1):81-91. doi: 10.1017/s0952523800003242.
The effects of small changes in the calcium and sodium concentrations and in the pH of superfusion medium on the membrane potential and light-evoked responses of cone horizontal cells in the goldfish retina were examined. Conventional intracellular recording, a bicarbonate-based superfusion medium, and a specially designed superfusion apparatus that reduced pressure wave disturbances were used. An increase in the extracellular calcium concentration, [Ca2+]o, from control levels (0.1 mM) to 1.0 mM hyperpolarized cone horizontal cells and reduced the magnitude of their light responses at all stimulus intensities. Addition of 20 mM NaCl to the 1.0 mM Ca2+ Ringer's solution reversed the hyperpolarizing effect of the 1.0 mM Ca2+ but addition of 20 mM choline, a monovalent cation that does not pass through cyclic GMP-activated channels, did not. Reduction of the superfusate pH from 7.6 to 7.2 by switching from a Ringer's solution gassed with 3% CO2 to one gassed with 10% CO2 hyperpolarized horizontal cells and reduced the magnitude of their light responses at all stimulus intensities for both 0.1 and 1.0 mM Ca2+ Ringer's solutions. An increase in pH to 8.2 by gassing the superfusate with 1% CO2 slightly depolarized the cells in 0.1 mM Ca2+ Ringer's solution but slightly hyperpolarized the cells in the 1.0 mM Ca2+ Ringer's solution. Following pharmacological isolation of the horizontal cells from synaptic input with high doses of glutamate (4-5 mM) and/or Co2+ (4 mM) treatment, no effect on horizontal cell membrane potential due to changes in pHo or [Ca2+]o was observed. These findings are discussed with respect to the cellular mechanisms and sites of action in the outer retina that are affected by changes in pHo and [Ca2+]o.
研究了灌注液中钙、钠浓度及pH值的微小变化对金鱼视网膜视锥水平细胞的膜电位和光诱发反应的影响。采用传统的细胞内记录方法、基于碳酸氢盐的灌注液以及一种专门设计的可减少压力波干扰的灌注装置。细胞外钙浓度[Ca2+]o从对照水平(0.1 mM)增加到1.0 mM时,视锥水平细胞发生超极化,并在所有刺激强度下降低其光反应的幅度。向1.0 mM Ca2+林格氏液中添加20 mM NaCl可逆转1.0 mM Ca2+的超极化作用,但添加20 mM胆碱(一种不能通过环鸟苷酸激活通道的单价阳离子)则不能。通过将灌注液从用3% CO2通气的林格氏液切换为用10% CO2通气的林格氏液,使灌注液pH值从7.6降至7.2,对于0.1 mM和1.0 mM Ca2+林格氏液,视锥水平细胞均发生超极化,并在所有刺激强度下降低其光反应的幅度。通过用1% CO2给灌注液通气使pH值升至8.2,在0.1 mM Ca2+林格氏液中细胞略有去极化,而在1.0 mM Ca2+林格氏液中细胞略有超极化。在用高剂量谷氨酸(4 - 5 mM)和/或Co2+(4 mM)处理对视锥水平细胞进行突触输入的药理学隔离后,未观察到pHo或[Ca2+]o变化对视锥水平细胞膜电位的影响。结合细胞机制和外视网膜中受pHo和[Ca2+]o变化影响的作用位点对这些发现进行了讨论。