Summary
Cell death by apoptosis is now recognized widely as an important constituent of cell turnover and disease pathology. Characterized by the cleavage of DNA into oligonucleosome-sized DNA fragments, end-labeling of fragmented DNA often is used as an in situ histological marker of apoptosis. The judicious and appropriate use of this technique therefore provides us with an important tool for assessing cell kinetics. Protocols for both terminal transferase-mediated UTP nick end-labeling, so-called TUNEL, and the combination of TUNEL with immunohistochemical staining are presented here, along with a discussion of its significance and interpretation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ueda N. and Shah N. (1994) Apoptosis. J Lab. Clin. Med. 124, 169–177.
Martin D. A. and Elkon K.B. (2004) Mechanisms of apoptosis. Rheum. Dis. Clin. North Am. 30, 441–454.
Arends M. J., Morris R. G., and Wyllie A. H. (1990) Apoptosis; the role of endonuclease. Am J. Pathol. 136, 593–608.
Gavrieli Y., Shermm Y., and Ben-Sasson S. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119, 493–501.
Fekete D. M., Homburger S. A., Waring M. T., Riedl A. E., and Garcia L. F. (1997) Involvement of programmed cell death in morphogenesis of the vertebrate inner ear. Development 124, 2451–2461.
Smith K. G. C., Hewitson T. D., Nossal G. J. V., and Tarlinton D. M. (1996) The phenotype and fate of the antibody-forming cells of the splenic foci. Eur. J. Immunol. 26, 444–448.
Darby I. A., Bisucci T, Hewitson T. D., and MacLellan D. G. (1997) Apoptosis is increased in a model of diabetes-impaired wound healing in genetically diabetic mice. Int. J. Biochem. Cell Biol. 29, 191–200.
Baker A. J., Mooney A., Hughes I., Lombardi D., Johnson R. J., and Savill J. (1994) Mesangial cell apoptosis: the major mechanism for resolution of glomerular hypercellularity in experiments mesangial proliferative nephritis. J. Clin. Invest. 94, 2105–2116.
Ansari B., Coates P. J., Greenstein B. D., and Hall P. A. (1993) In situ end labeling detects DNA strand breaks in apoptosis and other physiological and pathological states. J. Pathol. 170, 1–8.
Hockenbery D. (1995) Defining apoptosis. Am. J. Pathol. 146, 16–19.
Hewitson T. D., Smith K. G. C., and Becker G. J. (1996) Apoptosis and resolution of experimental renal infective tubulointerstitial nephritis. Nephrology 2, 127–132.
Strater J., Gunthem A. R., Bruderlein S., and Mollery. (1995) Microwave irradiation of paraffin-embedded tissue sensitizes the TUNEL method for in situ detection of apoptotic cells. Histochemistry 103, 157–160.
Wijsrnan J. H., Jonker R. R., Keijzer R., Van de Velde C. J. H., Comelisse C. J., and Van Dierendonck J. H. (1993) A new method to detect apoptosis in paraffin sections: in situ end-labeling of fragmented DNA. J. Histochem Cytochem. 41, 7–12.
Gorczym W., Gong J., and Darzynkiewicz Z. (1993) Detection of DNA strand breaks in individual apoptotic cells by the in situ terminal deoxynucleotidyl transferase and nick translation assays. Cancer Res. 53, 1945–1951.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Hewitson, T.D., Bisucci, T., Darby, I.A. (2006). Histochemical Localization of Apoptosis With In Situ Labeling of Fragmented DNA. In: Darby, I.A., Hewitson, T.D. (eds) In Situ Hybridization Protocols. Methods in Molecular Biology™, vol 326. Humana Press. https://doi.org/10.1385/1-59745-007-3:227
Download citation
DOI: https://doi.org/10.1385/1-59745-007-3:227
Publisher Name: Humana Press
Print ISBN: 978-1-58829-402-9
Online ISBN: 978-1-59745-007-2
eBook Packages: Springer Protocols