Location and function of voltage-sensitive conductances in retinal rods of the salamander Ambystoma tigrinum.

摘要

The functional role and spatial location of voltage-sensitive conductances that modify the light-evoked electrical signals were studied in retinal rods of the tiger salamander. An isolated rod was drawn into a suction electrode for recording membrane current and impaled with an intracellular electrode for recording membrane potential and passing current. A bright flash gave a rapid initial hyperpolarization that relaxed to a smaller plateau. Simultaneously the dark current of the outer segment was shut off with the time course of a rounded step function. This characteristic difference between the wave forms demonstrates that the voltage relaxation does not result from reopening of light-sensitive channels. The voltage relaxation in (2) did not require light or interruption of the dark current, as the wave form was duplicated by suddenly switching off a depolarizing current injected during steady saturating light. This is explained if the relaxation depends purely on voltage-sensitive conductances. The voltage response to a dim flash reached its peak value before the current response. The voltage wave form was predicted assuming that the recorded photo-current drove a linear high-pass filter with parameters derived from analysis of the voltage response to injection of a current step. When the intracellular voltage was changed by current injection the slope resistance of the outer segment slowly declined to a lower level, indicating that the outer segment contains a voltage-sensitive conductance. When a current step was injected in bright steady light, the current recorded from the outer segment consisted of a capacity component proportional to dV/dt and a small extracellular leakage current but no detectable ionic current. This supports other evidence indicating that light-sensitive channels comprise the main or exclusive ionic conductance of the outer segment. The behaviour in (5) is explained if the light-sensitive channels themselves are slowly opened by hyperpolarization and closed by depolarization. Analysis of the current-injection experiments suggests that most of the high-pass filtering in a rod results from the action of voltage-sensitive conductances located in the inner segment. Addition of 10 mM-CsCl to the Ringer solution abolished the relaxation in the voltage response to a bright flash but left intact the high-pass filtering of small signals. This would be explained by a selective block of one of two sets of voltage-sensitive channels in the inner segment or by a voltage-sensitive block of one kind of channel.