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 ...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 rd1展开更多
Background Glaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project ...Background Glaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats. Methods Chronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas. Results Compared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation （（2317.41±29.96）/mm^2 vs （1815.82±24.25）/mm^2; （26.20±2.10）/mm^2 vs （20.62±1.52）/mm^2, respectively）. The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells. Conclusion Both melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.展开更多
基金Supported by the Chongqing Internationa Cooperation Key Projects(No.CSTC2013GJHZ10004)National Basic Research Program of China(973 Program No.2013CB967002)
文摘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 rd1
基金This study was supported by a grant from the National Natural Science Foundation of China （No. 30571991）.
文摘Background Glaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats. Methods Chronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas. Results Compared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation （（2317.41±29.96）/mm^2 vs （1815.82±24.25）/mm^2; （26.20±2.10）/mm^2 vs （20.62±1.52）/mm^2, respectively）. The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells. Conclusion Both melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.
