Tachibanaki S, Tsushima S, Kawamura S
Department of Biology, Graduate School of Science, Osaka University, Machikane-yama 1-1, Toyonaka, Osaka 560-0043, Japan.
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):14044-9. doi: 10.1073/pnas.241396898. Epub 2001 Nov 13.
Vertebrate cone photoreceptors are known to show lower light sensitivity and briefer photoresponses than rod photoreceptors. To understand the molecular mechanisms characterizing cone photoresponses, we compared some of the reactions in the phototransduction cascade between rods and cones. For this purpose, rods and cones were obtained in quantities large enough to do biochemical studies. The cells were purified from the retina of carp (Cyprinus carpio) with a stepwise Percoll gradient. The purified rod fraction contained almost no other kinds of cells besides rods, and the purified cone fraction contained a mixture of red-, green-, and blue-sensitive cones in the ratio 3: approximately 1: approximately 1. We prepared membrane preparations from the rod and the cone fraction, and in these membranes, we measured activation efficiencies of the reactions in the phototransduction cascade. The results showed that the signal amplification is lower in the cone membranes, which accounts for the lower light sensitivity in cones. Furthermore, we measured the time courses of visual pigment phosphorylation. The result showed that the phosphorylation is much faster in the cone membranes, which also explains the lower light sensitivity and, in addition, the briefer photoresponse in cones.
脊椎动物的视锥光感受器的光敏感度低于视杆光感受器,且光反应持续时间更短。为了解视锥光反应的分子机制,我们比较了视杆和视锥光转导级联反应中的一些反应。为此,获取了足够数量的视杆和视锥细胞以进行生化研究。通过连续的Percoll梯度从鲤鱼(Cyprinus carpio)视网膜中纯化细胞。纯化后的视杆部分除视杆细胞外几乎不包含其他类型的细胞,纯化后的视锥部分包含红敏、绿敏和蓝敏视锥细胞的混合物,比例约为3:1:1。我们从视杆和视锥部分制备了膜制剂,并在这些膜中测量了光转导级联反应中各反应的激活效率。结果表明,视锥细胞膜中的信号放大较低,这解释了视锥细胞光敏感度较低的原因。此外,我们测量了视色素磷酸化的时间进程。结果表明,视锥细胞膜中的磷酸化速度要快得多,这也解释了视锥细胞光敏感度较低以及光反应持续时间更短的原因。