Protein screening and analysis for the retina degeneration

通过蛋白质组学分析筛选与视网膜存活、分化和再生相关的蛋白，鉴定出转录因子、结构蛋白、凋亡因子、分化因子和新蛋白等多种蛋白。其中，我们重点研究了分化蛋白家族中的Wnt14，并对Wnt14的细胞信号传导进行了分析。免疫组化结果表明，Wnt14存在于鸡胚视网膜神经节细胞内。并且，过表达Wnt14可抑制谷氨酸神经元毒性所致大鼠视网膜细胞系R28细胞的细胞死亡。此外，Wnt14通过抑制活化的caspase-3具有神经元保护作用。细胞凋亡诱导物。另一方面，我们也采用新的porteomics方法，利用青光眼模型小鼠视网膜，寻找青光眼中与视网膜存活因子相关的蛋白。结果，确定了青光眼视网膜变性的蛋白靶点。其中一种鉴定蛋白PDGFR位于小鼠视网膜视网膜神经节细胞中，PDGFR特异性配体PDGF-AA对谷氨酸诱导的大鼠视网膜神经节细胞系RGC-5细胞死亡具有神经保护作用。最后，我们成功地发现了新的视网膜神经营养蛋白。此外，我们对青光眼患者的基因组DNA进行了检测，发现tnf - α、血管紧张素- ii受体、内皮素A受体多态性与青光眼相关。另外，在阿尔茨海默病中发现的ApoE多态性与青光眼无关

Proteomics analysis was performed for the screening of proteins, related with retina survival, differentiation and regeneration Many proteins, including transcriptional factors, structural proteins, apoptosis factor, differentiation factors and novel proteins, were identified. Among of them, we focus on the Wnt14, one of the protein family for the differentiation, and the cell signaling of Wnt14 was analyzed. Immunohistochemistry shows that Wnt14 was endogeneously located in retinal ganglion cells of chick embryo retina. And, overexpressed Wnt14 inhibit cell death of R28 cells, rat retinal cell line, induced by glutamate neuronal toxity. Moreover, Wnt14 have the neuronal protective effect through the inhibition of activated caspase-3. apoptosis inducer.On the other hand, we also newly porteomics, using glaucoma model mouse retina, to find out the proteins associated with retinal survival factor in glaucoma. In result, the protein targets for glaucomatous retina degeneration were identified. PDGFR, one of the identified protein, was located in retinal ganglion cells on mouse retina, and PDGF-AA, specific ligand for PDGFR, have the neuro protective effect on the cell death of RGC-5, rat retina ganglion cell line, induced by glutamate. Finally, we succeeded in finding out newly neurotrophic proteins for retina.In addition, we check the genomic DNA of glaucoma patients, and we found polymorphism of TNF-alpha, Angiotensin-II receptor, Endothelin A receptor were associated with glaucoma. Otherwise, ApoE polymorphism, seen in Alzheimer disease, have no relation with glaucoma