Karolinska Institutet and Dainippon Sumitomo Pharma Alzheimer Center, Department of Neurobiology, Care Sciences and Society, NVS, Karolinska Institutet, Huddinge, Sweden.
J Cell Mol Med. 2012 Aug;16(8):1686-700. doi: 10.1111/j.1582-4934.2011.01441.x.
It is evident that the symptoms of Alzheimer's disease (AD) are derived from severe neuronal damage, and especially pyramidal neurons in the hippocampus are affected pathologically. Here, we analysed the proteome of hippocampal neurons, isolated from post-mortem brains by laser capture microdissection. By using (18)O labelling and mass spectrometry, the relative expression levels of 150 proteins in AD and controls were estimated. Many of the identified proteins are involved in transcription and nucleotide binding, glycolysis, heat-shock response, microtubule stabilization, axonal transport or inflammation. The proteins showing the most altered expression in AD were selected for immunohistochemical analysis. These analyses confirmed the altered expression levels, and showed in many AD cases a pathological pattern. For comparison, we also analysed hippocampal sections by Western blot. The expression levels found by this method showed poor correlation with the neuron-specific analysis. Hence, we conclude that cell-specific proteome analysis reveals differences in the proteome that cannot be detected by bulk analysis.
很明显,阿尔茨海默病(AD)的症状源自严重的神经元损伤,特别是海马体中的锥体细胞受到病理影响。在这里,我们通过激光捕获显微切割分析了来自死后大脑的海马神经元的蛋白质组。通过使用(18)O 标记和质谱法,估计了 AD 和对照组中 150 种蛋白质的相对表达水平。许多鉴定出的蛋白质参与转录和核苷酸结合、糖酵解、热休克反应、微管稳定、轴突运输或炎症。在 AD 中表达变化最明显的蛋白质被选择进行免疫组织化学分析。这些分析证实了表达水平的改变,并在许多 AD 病例中显示出病理模式。为了比较,我们还通过 Western blot 分析了海马切片。通过这种方法发现的表达水平与神经元特异性分析相关性较差。因此,我们得出结论,细胞特异性蛋白质组分析揭示了通过批量分析无法检测到的蛋白质组差异。