Schmitz Y, Witkovsky P
Dept. of Ophthalmology, New York University Medical Center, NY 10016, USA.
J Neurosci Methods. 1996 Sep;68(1):55-60. doi: 10.1016/0165-0270(96)00070-2.
In order to study glutamate release from light responsive photoreceptors, we used an eyecup preparation treated with detergent and distilled water, which permitted removal of the inner retina. The remaining 'reduced' retina consists mainly of photoreceptors attached to the pigment epithelium. The viability of the preparation was established by exclusion of trypan blue, light and electron microscopic examination of the photoreceptor layer and by intracellular recordings from rods. The 'reduced' retina was superfused at 1 ml/h and overflow samples were analyzed for their glutamate content by a fluorimetric enzyme assay. We tested the response to dark and light adaptation and to treatment with 100 microM CdCl2. We found a baseline glutamate level in light-adapted preparation which was not affected by cadmium. Dark adaptation induced a 2-fold increase of glutamate release, which was completely blocked by cadmium.
为了研究光响应性光感受器释放谷氨酸的情况,我们使用了经去污剂和蒸馏水处理的眼杯标本,这样可以去除视网膜内层。剩余的“简化”视网膜主要由附着于色素上皮的光感受器组成。通过台盼蓝排斥试验、光感受器层的光镜和电镜检查以及对视杆细胞的细胞内记录来确定标本的活力。“简化”视网膜以1毫升/小时的速度进行灌流,通过荧光酶法分析溢流样本中的谷氨酸含量。我们测试了对暗适应和光适应以及用100微摩尔/升氯化镉处理的反应。我们发现光适应标本中的谷氨酸基线水平不受镉的影响。暗适应导致谷氨酸释放增加两倍,这被镉完全阻断。