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目的建立适合研究膜质分离后海马组织不同分区亚蛋白质组的实验方法。方法显微切割获得成年大鼠海马不同解剖分区,然后按溶解性不同使用顺序抽提法细胞质和细胞膜蛋白,双向电泳进行分离蛋白后,采用与质谱兼容的高敏感银染显示蛋白斑点,经专业图像软件分析处理评价电泳效果。结果海马各个分区组织中膜蛋白得到有效富集,其中海马齿状回富集效率达8倍,图谱中可以显现的总蛋白斑点数目比传统方法增多30%。结论建立了理想的海马组织分区亚蛋白质组图谱,对膜蛋白实现了有效的富集,可以用于研究不同解剖分区在蛋白水平上的差异,为学习记忆、中枢神经缺血缺氧性损伤等多种后继研究提供了平台。
Objective To establish a suitable experimental method for the study of subfamilies in different subdivisions of hippocampus after membranous separation. Methods Different regions of the hippocampus of adult rats were obtained by microdissection. Then, the cytoplasm and the cell membrane proteins were extracted by different order of use. The proteins were separated by two-dimensional electrophoresis. The protein spots were displayed by high sensitivity silver stain compatible with mass spectrometry. Image software analysis and evaluation of electrophoresis. Results The membrane proteins were effectively enriched in each sub-regional tissue of hippocampus. The efficiency of dentate gyrus enrichment in hippocampus reached 8 times. The number of total protein spots in the map increased by 30% compared with the traditional method. Conclusion The ideal sub-proteome map of hippocampal tissue subdivision has been established and the membrane protein has been effectively enriched, which can be used to study the differences in protein levels between different anatomical subdivisions. It is also useful for learning and memory, hypoxic ischemic injury of central nervous system A variety of follow-up research provides a platform.