MacLeish P R, Schwartz E A, Tachibana M
J Physiol. 1984 Mar;348:645-64. doi: 10.1113/jphysiol.1984.sp015131.
THe current suppressed by light, the generator current, was studied in solitary salamander (Ambystoma tigrinum) rod photoreceptors with the single-micropipette voltage-clamp technique. The effects of Ca, cyclic GMP, and voltage were measured while voltage- and Ca-activated currents of the inner segment were blocked with Co, Cs, and TEA (tetraethylammonium). The generator current was increased more than 5-fold by lowering the external Ca concentration from 1.5 mM to 10 microM. The generator current could be decreased approximately 1/2 by injecting Ca into an outer segment. Injection of EGTA quickly increased the generator current approximately 2-fold. After injection ceased, the increase was quickly reversed. The generator current could be increased more than 5-fold by injecting cyclic GMP or 8Br-cyclic GMP. Injection of protons, the pH buffer bicine (N,N-bis[2-hydroxyethyl]glycine), or GMP did not produce a change in the generator current. The current-voltage curve for the generator current was influenced by external Co: in 3 mM-Co the current-voltage curve had a negative resistance between -45 and -90 mV; in 0.1 mM-Co the current-voltage curve paralleled the voltage axis between -45 and -90 mV. The difference is attributed to a voltage-dependent block by Co. Susceptibility to the blocking action of Co was reduced by lowering internal or external Ca concentration, or by injecting cyclic GMP. When rods were bathed in a medium containing 7-100 microM-Ca, a step depolarization produced a time-dependent decline in current. Because the reversal potential remained constant, the decline is attributed to an inactivation. The extent of inactivation was reduced by increasing the concentration of external Ca or injecting cyclic GMP.
利用单微吸管电压钳技术,在蝾螈(虎纹钝口螈)单个视杆光感受器中研究了受光抑制的电流,即发生器电流。在用钴、铯和四乙铵(TEA)阻断内段电压和钙激活电流的同时,测量了钙、环鸟苷酸和电压的影响。将外部钙浓度从1.5 mM降至10 μM时,发生器电流增加了5倍以上。向视杆外段注入钙可使发生器电流降低约1/2。注入乙二醇双四乙酸(EGTA)可迅速使发生器电流增加约2倍。注射停止后,增加的电流迅速恢复原状。注入环鸟苷酸或8-溴环鸟苷酸可使发生器电流增加5倍以上。注入质子、pH缓冲剂N,N-双(2-羟乙基)甘氨酸或鸟苷酸不会使发生器电流发生变化。外部钴会影响发生器电流的电流-电压曲线:在3 mM钴中,电流-电压曲线在-45至-90 mV之间具有负电阻;在0.1 mM钴中,电流-电压曲线在-45至-90 mV之间与电压轴平行。这种差异归因于钴的电压依赖性阻断。降低内部或外部钙浓度或注入环鸟苷酸可降低对钴阻断作用的敏感性。当视杆浸泡在含有7-100 μM钙的介质中时,阶跃去极化会导致电流随时间下降。由于反转电位保持恒定,这种下降归因于失活。增加外部钙浓度或注入环鸟苷酸可降低失活程度。