Schmidt K F, Nöll G N, Jacobi P, Baumann C
Physiologisches Institut, Justus-Liebig-Universität, Giessen, Germany.
Graefes Arch Clin Exp Ophthalmol. 1994 Mar;232(3):153-61. doi: 10.1007/BF00176785.
The whole-cell patch-clamp technique was employed to investigate the light responses of single retinal rods of the frog (Rana esculenta and R. temporaria). In the majority of experiments, completely isolated cells were studied. Coupling with neighboring cells gave rise to a more complex response configuration. Responses were recorded under voltage-clamp and under current-clamp conditions. Stimulus response curves were measured in experiments with local stimuli illuminating only parts of the outer segment. Metabolic factors such as cGMP, GTP and ATP were also tested and were found to have specific and different influences on the response configurations. When the recording pipette was filled with an intracellular medium devoid of nucleotides, a retardation in the recovery of the light responses was observed during the course of an experiment. Addition of 1 mM ATP to the pipette medium prevented the larger part of the retardation, while 1 mM GTP accelerated the response recovery at the beginning of an experiment but did not prevent a subsequent retardation. Micromolar concentrations of cGMP were sufficient to elicit both a depolarization of the photoreceptor membrane and an increase in the response duration. These results show that, in single photoreceptors, the configuration of light responses not only depends on the stimulus parameters but also on those properties of the cells that are directly controlled by their nucleotide metabolism.
采用全细胞膜片钳技术研究青蛙(食用蛙和林蛙)单个视网膜视杆细胞的光反应。在大多数实验中,研究的是完全分离的细胞。与相邻细胞耦合会产生更复杂的反应模式。在电压钳和电流钳条件下记录反应。在仅照亮外段部分区域的局部刺激实验中测量刺激反应曲线。还测试了诸如环磷酸鸟苷(cGMP)、鸟苷三磷酸(GTP)和三磷酸腺苷(ATP)等代谢因子,发现它们对反应模式有特定且不同的影响。当记录微电极中充满不含核苷酸的细胞内介质时,在实验过程中观察到光反应恢复延迟。向微电极介质中添加1 mM ATP可防止大部分延迟,而1 mM GTP在实验开始时加速反应恢复,但不能防止随后的延迟。微摩尔浓度的cGMP足以引起光感受器膜去极化并延长反应持续时间。这些结果表明,在单个光感受器中,光反应模式不仅取决于刺激参数,还取决于细胞中那些直接受其核苷酸代谢控制的特性。