多巴胺系统在慢性高眼压视网膜Müller细胞激活和神经节细胞损伤中的作用及机制研究

多巴胺(dopamine, DA)系统在调控视网膜视觉信息处理过程中发挥有重要的作用，该系统也与多种视网膜疾病的发生发展密切相关，但其在青光眼病理中的作用和机制还是亟待研究的重要问题。我们在慢性高眼压动物模型上的初步实验结果显示，视网膜DA含量和D1受体的表达上调，并通过调控外向钾电流可能参与Müller胶质细胞的激活及神经节细胞的损伤过程。本课题拟在慢性高眼压模型，利用膜片钳电生理、免疫组织(细胞)化学、Western blot、玻璃体显微注射和分子生物学等多学科研究手段，在视网膜切片、完整视网膜铺片、急性分离的视网膜细胞、纯化培养的Müller细胞等上，研究DA系统在慢性高眼压大鼠(或小鼠)视网膜的动态变化，分析其在Müller细胞激活和神经节细胞损伤中的作用及机制。本课题的研究成果将为阐明内源性DA系统在青光眼视网膜细胞损伤的作用及机制，以及有效地防治青光眼提供理论参考依据。

Dopamine (DA) and its receptor signaling play important roles in the processing of retinal visual information. Dopaminergic system is also involved in the pathology and pathphysiology of diverse retinal diseases. However, the effects and mechanisms of dopaminergic system in glaucoma are largely unknown. Our preliminary results obtained from a rat chronic ocular hypertension model showed that DA contents and D1 receptor expression were up-regulated, which may modulate outward K+ currents of retinal cells, thus involving in retinal Müller cell gliosis and ganglion cell injury. In the present proposal, by using patch-clamp recordings, immunohistochemistry (immunocytochemistry), Western blot, intravitreal microinjection and molecular biology techniques, we aim to explore the changes in DA contents and DA receptor expression in the retinal cells in a rat chronic ocular hypertension (COH) model, and to investigate the modulatory effects dopaminergic signaling system on Müller cell gliosis and ganglion cell injury, and the underlying mechanisms. The experiments will be done in retinal slices, whole-mounted retinal preprations, purified cultured retinal Müller cells, acutely isolated Müller cells and ganglion cells. Our prospective research achievements would provide a scientific basis for illuminating the mechanisms of retinal ganglion cell injury and for effctively prevention and cure of glaucoma.