Localised protein expression changes in different regions of human Alzheimer’s brain correlate with disease severity - Cerebellum

Human brain tissue was obtained from the New Zealand Brain Bank from donors with Alzheimer's disease (AD) or age-matched controls (n=9/group). Mean post mortem delays were ~12h. Six distinct brain regions were dissected, Hippocampus, Cingulate Gyrus, Entorhinal Cortex (severely affected), Motor and Sensory Cortex (less affected) and Cerebellum (spared). Protein was extracted from each sample and, each region was analysed independently. For each region, samples were divided into 3 sets of 3 cases and 3 controls and assigned to an iTRAQ 8 plex, alongside two aliquots of a pooled QC sample. Samples were digested, iTRAQ labelled and analysed. For each region, between 3000 and 4200 proteins were compared between AD and control, identifying a series of known and novel protein expression changes associated with the progression of AD.

本研究所用的人脑组织取自新西兰脑库，供体为阿尔茨海默病（Alzheimer's Disease, AD）患者或年龄匹配的健康对照者，每组样本量为9例。平均死后间隔时间约为12小时。共解剖分离6个不同脑区：海马体（Hippocampus）、扣带回（Cingulate Gyrus）、内嗅皮层（Entorhinal Cortex，受累严重）、运动与感觉皮层（Motor and Sensory Cortex，受累较轻）以及小脑（Cerebellum，未受累）。从每份样本中提取蛋白质，并对每个脑区独立开展分析。针对每个脑区，将样本分为3组，每组包含3例AD患者样本与3例对照样本，并与两份混合质控样品（pooled QC sample）的等分试样一同分配至iTRAQ 8 plex标记体系。随后对样本进行酶解、iTRAQ标记并完成检测分析。针对每个脑区，在AD患者与对照样本间对比了3000至4200种蛋白质，鉴定出一系列与AD进展相关的已知及新型蛋白质表达差异。