Rat retina and optic nerve LC-MS/MS

Various retinal disorder such as glaucomatous, retinal ischemia reperfusion, and traumatic optic neuropathy, are involved in the pathogenesis of neurodegeneration via glutamate exicitotoxicity. However, the proteomic characteristics and modulation of the neural-microenvironment with NMDA-induced neurodegeneration in retina and optic nerve remain partly understood. We established a protein sketch of NMDA-induced injury by comparing the proteomes of the PBS-operated, NMDA-operated and control groups. We carried out mass spectrometry-based label-free quantitative proteomics to investigate the exicitotoxic neurodegeneration mechanisms and identify key proteins that regulated neural cell death related signaling pathway in retina and optic nerve spatially. Using LC-MS/MS proteomics analysis, in total, we identified 3532 proteins in retina, 2593 proteins in optic nerve. According to Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), the protein changes and energy metabolism in retina and optic nerve tissue were comprehensively evaluated.

多种视网膜疾病，如青光眼性视网膜病变、视网膜缺血再灌注损伤、创伤性视神经病变等，均可通过谷氨酸兴奋性毒性参与神经退行性病变的发病过程。然而，针对视网膜与视神经中N-甲基-D-天冬氨酸（NMDA）诱导的神经退行性病变，其神经微环境的蛋白质组学特征及调控机制目前仍未完全阐明。本研究通过对比磷酸盐缓冲液（PBS）处理组、NMDA处理组与对照组的蛋白质组，构建了NMDA诱导损伤的蛋白质表达图谱。本研究采用基于质谱的无标记定量蛋白质组学技术，探究谷氨酸兴奋性毒性介导的神经退行性病变机制，并在空间维度上筛选出调控视网膜与视神经中神经细胞死亡相关信号通路的关键蛋白。通过液相色谱-串联质谱（LC-MS/MS）蛋白质组学分析，本研究共在视网膜组织中鉴定出3532种蛋白质，在视神经组织中鉴定出2593种蛋白质。本研究基于基因本体（Gene Ontology，GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）数据库，对视网膜与视神经组织的蛋白质表达变化及能量代谢情况进行了全面评估。