Study Of CD3ζ Gene On Retinal Ganglion Cells In Mice Of Chronic High Intraocular Pressure Model

BackgroundGlaucoma is one of the world’s one of the major irreversible blinding eye disease,caused by peripheral visual field defect until central vision disappeared. Retinal ganglion cells(retinal ganglion cell, RGC) apoptosis is one of the main features of glaucoma, but the specific mechanism which RGC injury is unknown. RGC damage is a worldwide problem, how to reduce glaucoma observation RGC lost speed and the change of the form to reveal glaucoma progression has important clinical significance. Traditional research methods of RGC damage cannot live dynamic monitoring the change of the RGC, current international in vivo confocal laser microscope to observe the transgenic mice with spontaneous fluorescence RGC change is a new trend, but mainly in the observation of RGC quantity changes, recent studies have shown that the possibility of living watch monkey and mouse RGC dendritic [1,2], but there’s no research involves the pathological state, the observation of the RGC morphological changes. Elevated intraocular pressure, intraocular pressure, IOP) is one of the main reasons for the onset of glaucoma, but can not explain all the characteristics that the onset of glaucoma [3-6]. Immune system involved in the pathogenesis of glaucoma in recent years, research has become more and more attention, previous research has show that CD3is the key factor of RGC dendritic morphology and function of dendritic CD3mice density was significantly greater than age [7] wild type mice. CD3knockout mice RGC dendritic cells increased obviously, CD3affected the RGC dendritic morphology and function of physiological condition, but is pathological condition caused the RGC injury needs further research.PurposesTo establish animal model of glaucoma vivo observation RGC change to choose the appropriate model, then observe the changes of RGC ischemia-reperfusion injury cases, the CD3relationship with the RGC damage. Methods1Three kinds of glaucoma model:1:1) the optic nerve contusion model can use automatic closing tweezers task of optic nerve;2) burning on venous vascular network of limbus of cornea and sclera in mice induced by chronic high intraocular pressure model;3) laser photocoagulation limbal induced chronic high intraocular pressure model in mice.2RGC damage observation:in vivo confocal laser microscope dynamic observation Thyl-CFP, the number of RGC mice with spontaneous fluorescence changes.3dual-channel excitation by confocal laser microscope observation ischemia-reperfusion injury cases Thyl-CFP RGC in mice.4Brn3b positive RGC detection:glaucoma model is set up, take out the whole retina retina shop, counting Brn3b positive RGC.Results1set up three kinds of glaucoma model:1) model of optic nerve injury;2) burning on venous vascular network of limbus of cornea and sclera in mice induced by chronic high intraocular pressure model;3) laser photocoagulation limbal induced chronic high intraocular pressure model.2optic nerve contusion model RGC reduce concentrated on postoperative week,7days RGC decreased by96.8%(P<0.001), burning on the vascular network of limbus of cornea and sclera in mice induced by intravenous RGC reduce concentrated on postoperative day,24hours RGC decreased by94.7%(P<0.001), decrease of RGC chronic high intraocular pressure continues to slow,4weeks RGC decreased by25.0%(P<0.001).3with Thyl-CFP chronic high intraocular pressure model in mice, live observation results show that the mice Thyl-CFP RGC reduce22%plus or minus4.8%after3weeks (p<0.001), six weeks after RGC reduce30%plus or minus4.7%(p<0.001).4ischemia-reperfusion injury causes Thyl-CFP RGC continued death in mice, CFP fluorescent protein is to observe the early RGC change effective tool. 5CD3zeta knockout in the chronic high intraocular pressure model in mice had no significant effect of RGC apoptosis and reduce and role.Conclusion1optic nerve contusion model caused by RGC reduce mainly concentrated in the postoperative week, burning on the vascular network of limbus of cornea and sclera in mice induced by chronic high intraocular pressure model induced by intravenous RGC reduce concentrated on postoperative24hours, laser photocoagulation induced corneal limbus RGC reduce chronic high intraocular pressure model is continuous slow, more fitting the clinical state of glaucoma.2available vivo confocal laser microscope observation of chronic high intraocular pressure model has been established Thyl-CFP the number of RGC mice decrease loss.3ischemia-reperfusion injury model is a kind of mature experimental animal models of neurodegenerative, in this study by looking at different time points after reperfusion RGCs of CFP expression can clearly see RGC apoptosis and reduces the number of trend. By observing Thyl-CFP autofluorescence protein compared to other methods of color tag may be observed in neurodegenerative diseases RGC’s early death is more advantage, and more sensitive.4compared with wild type mice, CD3zeta knockout mice of RGC damage in the chronic high intraocular pressure conditions did not significantly reduce, prompt CD3zeta may has nothing to do with the RGC damage.