Ripps H, Shakib M, MacDonald E D
J Cell Biol. 1976 Jul;70(1):86-96. doi: 10.1083/jcb.70.1.86.
The photoreceptors of dark-adapted skate retinas bathed in a Ringer solution containing horseradish peroxidase (HRP) incorporate the tracer into membrane-bound compartments within the synaptic terminal of the cell; after 1 or 2 h of incubation, approx. 10-38% of the synaptic vesicles were labeled. The receptors appeared to be functioning normally throughout the incubation period, since electrical potentials of normal amplitude could be elicited in response to dimphotic stimuli. However, it was possible to block the uptake of peroxidase by a regimen of light adaptation that effectively suppressed light-induced activity in the electroretinogram. If, during incubation with peroxidase, retinas were exposed at 10-min intervals to an intense 1-ms flash from a xenon discharge tube, the receptor terminals were almost completely devoid of peroxidase; fewer than 2% of the vesicles were labeled. The suppression of HRP uptake could also be achieved in dark-adapted retinas by adding magnesium to the bathing solution, suggesting that calcium is necessary for transmitter release from vesicles in the receptor terminals. These findings are consistent with the view that vertebrate photoreceptors discharge a neurotransmitter in darkness, and that light decreases the release of this substance. It seems likely that the incorporation of peroxidase into vesicles of physiologically active receptor terminals reflects a mechanism for the retrieval of vesicle membrane after exocytosis.
将暗适应的鳐鱼视网膜的光感受器置于含有辣根过氧化物酶(HRP)的林格氏液中,该酶会被细胞突触终末内的膜结合区室摄取;孵育1或2小时后,约10% - 38%的突触小泡被标记。在整个孵育期间,感受器似乎功能正常,因为对弱光刺激能引发正常幅度的电位。然而,通过一种能有效抑制视网膜电图中光诱导活动的光适应方案,可以阻断过氧化物酶的摄取。如果在与过氧化物酶孵育期间,视网膜每隔10分钟暴露于氙放电管发出的1毫秒强光闪光下,感受器终末几乎完全没有过氧化物酶;标记的小泡不到2%。通过向浴液中添加镁,在暗适应的视网膜中也能实现对HRP摄取的抑制,这表明钙对于感受器终末小泡释放神经递质是必需的。这些发现与脊椎动物光感受器在黑暗中释放神经递质以及光会减少这种物质释放的观点一致。过氧化物酶掺入生理活性感受器终末的小泡中,似乎反映了胞吐作用后小泡膜回收的一种机制。