Thomas L B, Gates D J, Richfield E K, O'Brien T F, Schweitzer J B, Steindler D A
Department of Anatomy and Neurobiology, University of Tennessee, Memphis 38163, USA.
Exp Neurol. 1995 Jun;133(2):265-72. doi: 10.1006/exnr.1995.1029.
Deoxyribonucleic acid of cells undergoing apoptosis is cleaved by a calcium-dependent endonuclease into oligonucleosomal-sized fragments. These fragments can be labeled using the enzyme terminal deoxynucleotidyl transferase so that the cells can be visualized immunohistochemically. Few investigators have evaluated this method in disease processes of the human central nervous system. The Tdt-mediated dUTP-biotin nick end labeling (TUNEL) technique has been investigated in preliminary studies of a variety of pathologic conditions of the human brain (e.g., gliomas, traumatic brain injury, Parkinson's disease, Parkinson's-Alzheimer's complex, multisystem atrophy, striatonigral degeneration). We focus, however, on Huntington's disease (HD) because of the availability of well-characterized pathological stages for study, and also because of the neurodegenerative diseases studied to date, only Huntington's disease revealed significant and consistent labeling with this method. This implies a possibly unique nature to the mechanism of cell death in Huntington's disease compared to the other neurodegenerative diseases studied. TUNEL+ neurons were found in Grade 1-4 HD neostriatum, while labeled astrocytes were found predominantly in the Grade 1 and 2 cases studied to date. TUNEL+ cells were also found in glioblastoma multiforme and traumatic brain injury. We conclude that while there appear to be several limitations associated with this technique, it may be useful for identifying both apoptosis and necrosis in certain neuropathological conditions.
正在经历凋亡的细胞的脱氧核糖核酸被一种钙依赖性核酸内切酶切割成寡核小体大小的片段。这些片段可以用末端脱氧核苷酸转移酶进行标记，以便能够通过免疫组织化学方法观察到这些细胞。很少有研究者在人类中枢神经系统疾病过程中评估这种方法。在对人类大脑的多种病理状况（如神经胶质瘤、创伤性脑损伤、帕金森病、帕金森 - 阿尔茨海默综合征、多系统萎缩、纹状体黑质变性）的初步研究中，已经对Tdt介导的dUTP - 生物素缺口末端标记（TUNEL）技术进行了研究。然而，我们关注亨廷顿舞蹈病（HD），因为有特征明确的病理阶段可供研究，而且在迄今为止所研究的神经退行性疾病中，只有亨廷顿舞蹈病用这种方法显示出显著且一致的标记。这意味着与其他所研究的神经退行性疾病相比，亨廷顿舞蹈病细胞死亡机制可能具有独特的性质。在1 - 4级HD新纹状体中发现了TUNEL + 神经元，而到目前为止，标记的星形胶质细胞主要在1级和2级病例中发现。在多形性胶质母细胞瘤和创伤性脑损伤中也发现了TUNEL + 细胞。我们得出结论，虽然这种技术似乎存在一些局限性，但它可能有助于在某些神经病理状况下识别凋亡和坏死。
