Morphological and biochemical characterization and analysis of apoptosis
Morphological and biochemical characterization and analysis of apoptosis
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DOI：
10.1016/s1056-8719(97)00033-6
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发表时间：
1997-06-01
影响因子：
1.9
通讯作者：
Agrawal, DK
中科院分区：
文献类型：
--
作者：
Allen, RT;Hunter, WJ;Agrawal, DK
细胞凋亡是细胞程序性死亡的一种形式，具有生理和稳态功能。然而，最近的证据表明，细胞凋亡在已知的人类疾病，如心脏病，癌症，艾滋病，神经退行性疾病和自身免疫性疾病的病因学和病理生理学不断浮出水面。这就需要确定哪些方法是最有效的，并被广泛接受，以破译其在各种研究环境中的存在。因此，我们已经详细的形态学和生物化学特征的凋亡细胞死亡，重点是区分坏死。此外，我们描述了特定的和选择性的技术，这是最佳的目标标志性细胞凋亡的功能，如显微镜，膜联蛋白V标记，原位缺口末端标记（TUNEL），和DNA片段化分析凝胶电泳和ELISA法对DNA的大小。每种技术的优点和缺点进行了讨论，以及他们的实验相对于彼此的重要性。已经以逐步的方式描述了这些方法，并且可以容易地应用于大多数细胞系统。无论是在组织还是单细胞水平上工作，这些方法在定性和定量细胞凋亡方面都非常有效。这些方法与分子生物学技术相结合的应用可以进一步阐明细胞凋亡的潜在机制。(C)1997年爱思唯尔科学公司
Apoptosis is a form of programmed cell death serving physiologic and homeostatic functions. However, recent evidence implicating apoptosis in the etiology and pathophysiology of known human diseases, such as heart diseases, cancer, AIDS, and neurodegenerative and autoimmune diseases are continually surfacing. This has spawned the need for identifying which methods are the most effective and well accepted to decipher its presence in a variety of research settings. We have therefore detailed the morphology and biochemical features of apoptotic cell death, with an emphasis on discriminating it from necrosis. In addition, we describe specific and selective techniques which are optimal to target hallmark apoptotic features, such as microscopy, Annexin V labeling, in situ nick-end labeling (TUNEL), and DNA fragmentation analysis by gel electrophoresis and ELISA for oligonucleosome-sized DNA. The advantages and disadvantages of each technique are discussed, as well as their experimental importance relative to one another. The methods have been described in a stepwise fashion, and can readily be applied in the majority of cell systems. Whether working on the tissue or single cell level, these methods are highly effective in qualifying and quantifying apoptosis. The application of these methods in conjunction with molecular techniques can further delineate the underlying mechanisms of apoptosis. (C) 1997 Elsevier Science Inc.