INSITU END-LABELING DETECTS DNA STRAND BREAKS IN APOPTOSIS AND OTHER PHYSIOLOGICAL AND PATHOLOGICAL STATES
INSITU END-LABELING DETECTS DNA STRAND BREAKS IN APOPTOSIS AND OTHER PHYSIOLOGICAL AND PATHOLOGICAL STATES
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DOI：
10.1002/path.1711700102
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发表时间：
1993-05-01
影响因子：
7.3
通讯作者：
HALL, PA
中科院分区：
文献类型：
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作者：
ANSARI, B;COATES, PJ;HALL, PA
我们已经研究了使用一种新的技术，原位末端标记，作为一种手段，在福尔马林固定，石蜡处理的组织切片中的凋亡细胞的特异性识别。该技术依赖于凋亡细胞中DNA链断裂的存在，这是由细胞死亡过程中内源性核酸酶活性的激活引起的。在DNA聚合酶存在下，用非同位素报告分子标记这些链，并以化学方法鉴定标记的DNA。我们发现，原位末端标记染色细胞的形态特征的凋亡，并大大简化了他们的鉴定。此外，在两个模型系统中，标记细胞的数量与通过替代技术测量的经历凋亡的细胞的数量平行。DNA聚合酶的Klenow片段标记凋亡细胞核的能力表明，在此过程中观察到的特征性DNA片段化涉及形成具有5'突出端的DNA断裂。原位末端标记将是有价值的，在一系列正常组织和各种病理状态的细胞凋亡的识别和定量。然而，该技术并不特异于程序性细胞死亡，结果必须谨慎解释，并与凋亡的形态学标准相关。
We have investigated the use of a novel technique, in situ end-labelling, as a means of the specific identification of apoptotic cells in formalin-fixed, paraffin-processed tissue sections. The technique relies on the presence of DNA strand breaks in apoptotic cells, caused by activation of endogenous nuclease activity during the process of cell death. These strands are labelled with a non-isotopic reporter molecule in the presence of a DNA polymerase, and labelled DNA is identified immunohistochemically. We show that in situ end-labelling stains cells with the morphological characteristics of apoptosis, and greatly simplifies their identification. Furthermore, in two model systems, the number of labelled cells parallels the number of cells undergoing apoptosis as measured by alternative techniques. The ability of the Klenow fragment of DNA polymerase to label apoptotic nuclei suggests that the characteristic DNA fragmentation seen during this process involves the formation of DNA breaks with a 5' overhang. In situ end-labelling will be valuable for the identification and quantitation of apoptosis in a range of normal tissues and in a variety of pathological states. However, the technique is not specific for programmed cell death, and results must be interpreted with caution and correlated with morphological criteria of apoptosis.