False-positive apoptosis signal in mouse kidney and liver detected with TUNEL assay.
Apoptosis is a physiological, programmed process for the elimination of cells from living organisms. Currently, one of the most frequently used methods to detect apoptosis is TUNEL assay. It has provided valuable information about apoptosis in various tissues. However, the sensitivity and the specificity of TUNEL technique have also been criticized. We detected an intense false-positive apoptotic signal in nude and Balb/c mice kidney and liver. In kidney the signal was confined to the proximal, distal and collecting tubular cells, and in liver to hepatocytes. Both tissues appeared normal in light microscopy, and no DNA ladder formation or increase in caspase-3 enzyme activity was detected. BrdU labelling and Ki-67 immunostaining did not reveal increased cell proliferation in these tissues. On the other hand, false-positive signal was not detected in testis, spleen, pancreas or renal cell carcinoma from the same animals. Also, no false-positive signal was seen in human liver or kidney samples. Although factors known to produce false-positive staining related to sample harvesting, preparation and staining protocols were eliminated, the cause of the false- positive apoptotic signal remains unknown. We conclude that caution must be exercised when examining apoptosis in mouse tissues with TUNEL assay.
Department of Molecular Medicine, A.I.Virtanen Institute, Kuopio University Hospital, University of Kuopio, Finland.
K J
KJ
Pulkkanen
M O
MO
Laukkanen
J
J
Naarala
S
S
Yla-Herttuala
D000818:Animals; D017209:Apoptosis; D001973:Bromodeoxyuridine / Q000378:metabolism; D002292:Carcinoma, Renal Cell / Q000473:pathology; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002455:Cell Division; D004247:DNA / Q000378:metabolism; D053938:DNA Fragmentation; D005189:False Positive Reactions; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D007680:Kidney Neoplasms / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D012680:Sensitivity and Specificity; D013154:Spleen / Q000166:cytology; D013737:Testis / Q000166:cytology; D014407:Tumor Cells, Cultured / Q000637:transplantation
D004247:DNA; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D001973:Bromodeoxyuridine