Niemeyer G, Jurklies B, Kaelin-Lang A, Bittiger H
Neurophysiologie-Labor, Universitäts-Augenklinik Zürich.
Klin Monbl Augenheilkd. 1995 May;206(5):380-3. doi: 10.1055/s-2008-1035469.
Based upon the extensive information from various laboratories on cholinergic enzymes and receptors in mammalian retina as well as cholinergic effects on retinal neurons we became interested in studying (1) retinal binding of a muscarinic antagonist, Quinuclidinyl benzilate (QNB) and (2), effects of the antagonist on retinal information processing.
Eyes from deeply anesthetized cats were used for homogenate of freshly isolated retina in the binding study, or they were arterially perfused for electrophysiology in vitro. The electroretinogram (ERG) and the compound action potential of the optic nerve (optic nerve response, ONR) were recorded under rod- and cone-stimulating conditions. QNB was infused intraarterially for 10-30 min, followed by washout (avoiding recycling or extraocular metabolism).
3H-QNB revealed a high affinity to muscarinic receptors with a dissociation constant KD of 0.27 nM and a relatively high density of muscarinic binding sites of 110 fmol per mg protein. QNB enhanced the amplitude of the ERG b-wave, but decreased dose-dependently and reversibly the components of the ONR. In addition, we observed a moderate vasoconstriction as indicated by a slight, dose-related decrease in flow rate of perfusion.
The biochemical data on binding of 3H-QNB in connection with the marked electrophysiological changes induced by QNB suggest a substantial contribution of muscarinic cholinergic transmission in the cat retina.
基于各实验室提供的关于哺乳动物视网膜中胆碱能酶和受体的大量信息,以及胆碱能对视网膜神经元的作用,我们开始对以下研究感兴趣:(1)毒蕈碱拮抗剂苯甲托品(QNB)与视网膜的结合;(2)该拮抗剂对视网膜信息处理的影响。
在结合研究中,使用深度麻醉猫的眼睛制备新鲜分离视网膜的匀浆,或者将其用于体外电生理学研究的动脉灌注。在视杆和视锥细胞刺激条件下记录视网膜电图(ERG)和视神经复合动作电位(视神经反应,ONR)。通过动脉内注入QNB 10 - 30分钟,然后冲洗(避免循环或眼外代谢)。
³H-QNB显示出对毒蕈碱受体具有高亲和力,解离常数KD为0.27 nM,毒蕈碱结合位点的密度相对较高,为每毫克蛋白质110 fmol。QNB增强了ERG b波的振幅,但剂量依赖性且可逆地降低了ONR的成分。此外,我们观察到适度的血管收缩,表现为灌注流速略有剂量相关的降低。
³H-QNB结合的生化数据以及QNB引起的明显电生理变化表明,毒蕈碱胆碱能传递在猫视网膜中起重要作用。