Yang Dun-Sheng, Lee Ju-Hyun, Nixon Ralph A
Center for Dementia Research, Nathan S. Kline Institute, Orangeburg, New York, USA.
Methods Enzymol. 2009;453:111-44. doi: 10.1016/S0076-6879(08)04006-8.
This chapter describes detailed methods to monitor autophagy in neurodegenerative disorders, especially in Alzheimer's disease. Strategies to assess the competence of autophagy-related mechanisms in disease states ideally incorporate analyses of human disease and control tissues, which may include brain, fibroblasts, or other peripheral cells, in addition to animal and cell models of the neurodegenerative disease pathology and pathobiology. Cross-validation of pathophysiological mechanisms in the diseased tissues is always critical. Because of the cellular heterogeneity of the brain and the differential vulnerability of the neural cells in a given disease state, analyses focus on regional comparisons of affected and unaffected regions or cell populations within a particular brain region and include ultrastructural, immunological, and cell and molecular biological approaches.
本章介绍了监测神经退行性疾病,尤其是阿尔茨海默病中自噬的详细方法。评估疾病状态下自噬相关机制能力的策略理想情况下应纳入对人类疾病组织和对照组织的分析,这些组织可能包括大脑、成纤维细胞或其他外周细胞,此外还包括神经退行性疾病病理和病理生物学的动物和细胞模型。对患病组织中病理生理机制的交叉验证始终至关重要。由于大脑的细胞异质性以及特定疾病状态下神经细胞的不同易损性,分析重点在于特定脑区内受影响和未受影响区域或细胞群体的区域比较,并包括超微结构、免疫学以及细胞和分子生物学方法。