Human melanopsin-AAV2/8 transfection to retina transiently restores visual function in rd1 mice  

作　　者：Ming-Ming Liu Jia-Man Dai Wen-Yi Liu Cong-Jian Zhao Bin Lin Zheng-Qin Yin 

机构地区：[1]Southwest Hospital, Southwest Eye Hospital, Third Military Medical University, Chongqing 400038, China [2]Key Lab of Visual Damage and Regeneration and Restoration of Chongqing, Chongqing 400038, China [3]Departments of Anatomy and Ophthalmology, Li Ka Shing Faculty of Medicine, the University of Hong Kong, Hong Kong 200131, China

出　　处：《International Journal of Ophthalmology(English edition)》2016年第5期655-661,共7页国际眼科杂志（英文版）

基　　金：Supported by the Chongqing Internationa Cooperation Key Projects(No.CSTC2013GJHZ10004);National Basic Research Program of China(973 Program No.2013CB967002)

摘　　要：AIM： To explore whether ectopic expression of human melanopsin can effectively and safely restore visual function in rd1 mice.· METHODS： Hematoxylin-eosin staining of retinal sections from rd1 mice was used to detect the thickness of the outer nuclear layer to determine the timing of surgery. We constructed a human melanopsinAAV2/8 viral vector and injected it into the subretinal space of rd1 mice. The Phoenix Micron IV system was used to exclude the aborted injections, and immunohistochemistry was used to validate the ectopic expression of human melanopsin. Furthermore, visual electrophysiology and behavioral tests were used to detect visual function 30 and 45 d after the injection. The structure of the retina was compared between the human melanopsin-injected group and phosphate buffer saline（PBS）-injected group.·RESULTS： Retinas of rd1 mice lost almost all of their photoreceptors on postnatal day 28（P28）. We therefore injected the human melanopsin-adeno-associated virus（AAV） 2/8 viral vector into P30 rd1 mice. After excluding aborted injections, we used immunohistochemistry of the whole mount retina to confirm the ectopic expression of human melanopsin by co-expression of human melanopsin and YFP that was carried by a viral vector. At30 d post-injection, visual electrophysiology and the behavioral test significantly improved. However,restoration of vision disappeared 45 d after human melanopsin injection. Notably, human melanopsin-injected mice did not show any structural differences in their retinas compared with PBS-injected mice.·CONCLUSION： Ectopic expression of human melanopsin effectively and safely restores visual function in rd1AIM： To explore whether ectopic expression of human melanopsin can effectively and safely restore visual function in rd1 mice.· METHODS： Hematoxylin-eosin staining of retinal sections from rd1 mice was used to detect the thickness of the outer nuclear layer to determine the timing of surgery. We constructed a human melanopsinAAV2/8 viral vector and injected it into the subretinal space of rd1 mice. The Phoenix Micron IV system was used to exclude the aborted injections, and immunohistochemistry was used to validate the ectopic expression of human melanopsin. Furthermore, visual electrophysiology and behavioral tests were used to detect visual function 30 and 45 d after the injection. The structure of the retina was compared between the human melanopsin-injected group and phosphate buffer saline（PBS）-injected group.·RESULTS： Retinas of rd1 mice lost almost all of their photoreceptors on postnatal day 28（P28）. We therefore injected the human melanopsin-adeno-associated virus（AAV） 2/8 viral vector into P30 rd1 mice. After excluding aborted injections, we used immunohistochemistry of the whole mount retina to confirm the ectopic expression of human melanopsin by co-expression of human melanopsin and YFP that was carried by a viral vector. At30 d post-injection, visual electrophysiology and the behavioral test significantly improved. However,restoration of vision disappeared 45 d after human melanopsin injection. Notably, human melanopsin-injected mice did not show any structural differences in their retinas compared with PBS-injected mice.·CONCLUSION： Ectopic expression of human melanopsin effectively and safely restores visual function in rd1

关 键 词：human melanopsin retinal degenerative diseases visual restoration 

分 类 号：R774[医药卫生—眼科]