Department of Neuroanatomy, Max-Planck-Institute for Brain Research, D-60528 Frankfurt/Main, Germany.
J Neurosci. 2010 Jun 30;30(26):8745-58. doi: 10.1523/JNEUROSCI.4417-09.2010.
The progression of rod and cone degeneration in retinally degenerate (rd) mice ultimately results in a complete loss of photoreceptors and blindness. The inner retinal neurons survive and several recent studies using genetically targeted, light activated channels have made these neurons intrinsically light sensitive. We crossbred a transgenic mouse line expressing channelrhodopsin2 (ChR2) under the control of the Thy1 promoter with the Pde6b(rd1) mouse, a model for retinal degeneration (rd1/rd1). Approximately 30-40% of the ganglion cells of the offspring expressed ChR2. Extracellular recordings from ChR2-expressing ganglion cells in degenerated retinas revealed their intrinsic light sensitivity which was approximately 7 log U less sensitive than the scotopic threshold and approximately 2 log U less sensitive than photopic responses of normal mice. All ChR2-expressing ganglion cells were excited at light ON. The visual performance of rd1/rd1 mice and ChR2 rd1/rd1 mice was compared. Behavioral tests showed that both mouse strains had a pupil light reflex and they were able to discriminate light fields from dark fields in the visual water task. Cortical activity maps were recorded with optical imaging. The ChR2rd1/rd1 mice did not show a better visual performance than rd1/rd1 mice. In both strains the residual vision was correlated with the density of cones surviving in the peripheral retina. The expression of ChR2 under the control of the Thy1 promoter in retinal ganglion cells does not rescue vision.
Rod 和 cone 变性在视网膜变性(rd)小鼠中的进展最终导致光感受器完全丧失和失明。内视网膜神经元存活下来,最近的几项使用基因靶向、光激活通道的研究使这些神经元具有内在的光敏感性。我们将表达通道视紫红质 2(ChR2)的转基因小鼠与 Pde6b(rd1)小鼠(视网膜变性模型)杂交,后者在 Thy1 启动子的控制下表达 ChR2。大约 30-40%的后代神经节细胞表达 ChR2。从变性视网膜中表达 ChR2 的神经节细胞进行的细胞外记录显示了它们的内在光敏感性,大约比暗适应阈值低 7 个对数单位,比正常小鼠的明适应反应低 2 个对数单位。所有表达 ChR2 的神经节细胞在光 ON 时都会兴奋。比较了 rd1/rd1 小鼠和 ChR2 rd1/rd1 小鼠的视觉性能。行为测试表明,两种小鼠品系都有瞳孔光反射,并且能够在视觉水任务中区分明场和暗场。用光学成像记录皮质活动图。ChR2rd1/rd1 小鼠的视觉性能并不优于 rd1/rd1 小鼠。在两种品系中,残余视力与周边视网膜中存活的 cone 密度相关。视网膜神经节细胞中 Thy1 启动子控制下的 ChR2 表达不能挽救视力。