title: Apoptosis of cardiomyocytes in explanted and transplanted hearts. Comparison of results from in situ TUNEL, ISEL, and ISOL reactions.
abstract: We assessed the efficiency of detecting myocyte apoptosis within human hearts using in situ enzymatic reactions in paraffin-embedded tissue samples: in situ end labeling (ISEL), terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), and in situ oligoligation (ISOL). The reactions were carried out in explanted hearts (idiopathic dilatative cardiomyopathy, n = 6; ischemic heart disease, n = 3) and in endomyocardial biopsy specimens (EMBs; n = 32) obtained from transplanted human hearts. The results were verified by DNA laddering. The ISOL reaction led to a significantly (P = .027) smaller number of false-positive results (2/41 [5%]) compared with assessment by ISEL (9/41 [22%]) or TUNEL (9/41 [22%]). Only 1 ISEL+ apoptotic cardiomyocyte was found in specimens from explanted hearts. Among the EMBs, 1 specimens had TUNEL+ apoptotic cardiomyocytes and 1 specimen had ISEL+ apoptotic cardiomyocytes. This implies that verifying results by independent methods must be used for TUNEL and ISEL techniques. A smaller number of false-positive results makes interpretation of ISOL results easier, although the sensitivity of this reaction remains to be established.
authors.affiliation: Institute of Cardiology, Kaunas University of Medicine, Kaunas, Lithuania.
authors.forename: Vaiva
authors.initials: V
authors.lastname: Lesauskaite
authors.forename: Maria Carmela
authors.initials: MC
authors.lastname: Epistolato
authors.forename: Laima
authors.initials: L
authors.lastname: Ivanoviene
authors.forename: Piero
authors.initials: P
authors.lastname: Tanganelli
mesh_terms: D017209:Apoptosis; D002311:Cardiomyopathy, Dilated / Q000473:pathology* / Q000601:surgery; D002452:Cell Count; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation; D016027:Heart Transplantation; D006801:Humans; D007124:Immunoenzyme Techniques; D020287:In Situ Nick-End Labeling; D009206:Myocardium / Q000378:metabolism / Q000473:pathology*; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology*; D019737:Transplants
chemical_list: D004247:DNA