Differentiation between cellular apoptosis and necrosis by the combined use of in situ tailing and nick translation techniques.

摘要: Background A number of enzymatic techniques have recently been developed to detect DNA fragmentation in apoptosis at the cellular level. However, since also occurs necrosis, we studied which extent use polymerase (nick translation) or terminal transferase (tailing) allows differentiation between internucleosomal degradation, typical for apoptosis, and more random destruction necrosis. Experimental design We compared these on vitro vivo models apoptotic necrotic cell death. Apoptosis thymocytes was induced by gamma-irradiation, necrosis cytotoxic action antibody complement. Cell death examined paraffin-embedded tissue material from animals with autoimmune encephalomyelitis that served as a model kainic acid-induced nerve degeneration Results visualized incorporation labeled nucleotides into nuclei affected cells utilizing tailing nick translation techniques. In early stages vitro, undergoing were preferentially tailing, whereas identified translation. Similarly, detected translation, slightly sensitive detection apoptosis. obtained accord assessment morphologic criteria. Both could be applied samples even after prolonged fixation paraformaldehyde if sections pretreated proteinase K digestion. Conclusions Our studies show both situ are useful detecting suspensions sections. These may help define molecular mechanisms leading experimental conditions eventually human tissue.