10
TAKEHIKO KOJI
that of single-stranded DNA. That is why we have been using TdT
to label synthetic oligodeoxynucleotides as detailed in the protocol of in situ hybridization (Koji and Brenner 1993; Chapter 12
and 13). In fact, a specific nuclear staining was obtained by TUNEL in the frozen sections of CC14 treated liver, where almost all
of degenerating hepatocytes were observed with typical necrotic
features by electron microscopy (Hashimoto et al. 1995). In addition, Gold et al. (1994) reported that early stages of necrosis
were preferentially detected by ISNT, whereas TUNEL was
slightly more sensitive for the detection of apoptosis. In our
hands, it has turned out that TUNEL is specific enough to detect
apoptotic germ cells in mouse testis (Wang et al. 1998) and apopto tic renal tubular cells in ischemia-reperfusion mouse kidney
(Nogae et al. 1998), as verified by combining use of electron microscopy. Consequently, we should realize that the usefulness of
TUNEL in identifying apoptotic cells seems highly dependent
upon the tissue-type or cell-type to be analyzed. An example
of TUNEL staining of apoptotic spermatogenic cells in adult
mouse testis was shown in Fig. 4 (Koji and Wang 1998).
References
Appleby DW, Modak SP (1977) DNA degradation in terminally differentiating lens fiber cells from chick embryos. Proc Nat! Acad Sci USA
74:5579-5583
Boerrigter MET! (1991) DNA strand break metabolism in human lymphocytes: A reevaluation. Exp Cell Res 196:1-5
Bortner CD, Oldenburg NBE, Cidlowski JA (1995) The role of DNA fragmentation in apoptosis. Trends Cell BioI 5:21-26
Collins RJ, Harmon BV, Gobe GC, Kerr JFR (1992) Internucleosomal DNA
cleavage should not be the sole criterion for identifying apoptosis. Int J
Radiat Res 61:451-453
Dawson BA, Lough J (1988) Immunocytochemical localization of transient
DNA strand breaks in differentiating myotubes using in situ nick-translation. Dev BioI 127:362-367
Farzaneh F, Zalin R, Brill D, Shall S (1982) DNA strand breaks and ADPribosyl transferase activity in eukaryotic differentiation. Nature 300:362366
Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed
cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell
BioI 119:493-501
Gold R, Schmied M, Breitschopf H, Hartung HP, Toyka KV, Lassmann H
(1994) Differentiation between cellular apoptosis and necrosis by the
TAKEHIKO KOJI
that of single-stranded DNA. That is why we have been using TdT
to label synthetic oligodeoxynucleotides as detailed in the protocol of in situ hybridization (Koji and Brenner 1993; Chapter 12
and 13). In fact, a specific nuclear staining was obtained by TUNEL in the frozen sections of CC14 treated liver, where almost all
of degenerating hepatocytes were observed with typical necrotic
features by electron microscopy (Hashimoto et al. 1995). In addition, Gold et al. (1994) reported that early stages of necrosis
were preferentially detected by ISNT, whereas TUNEL was
slightly more sensitive for the detection of apoptosis. In our
hands, it has turned out that TUNEL is specific enough to detect
apoptotic germ cells in mouse testis (Wang et al. 1998) and apopto tic renal tubular cells in ischemia-reperfusion mouse kidney
(Nogae et al. 1998), as verified by combining use of electron microscopy. Consequently, we should realize that the usefulness of
TUNEL in identifying apoptotic cells seems highly dependent
upon the tissue-type or cell-type to be analyzed. An example
of TUNEL staining of apoptotic spermatogenic cells in adult
mouse testis was shown in Fig. 4 (Koji and Wang 1998).
References
Appleby DW, Modak SP (1977) DNA degradation in terminally differentiating lens fiber cells from chick embryos. Proc Nat! Acad Sci USA
74:5579-5583
Boerrigter MET! (1991) DNA strand break metabolism in human lymphocytes: A reevaluation. Exp Cell Res 196:1-5
Bortner CD, Oldenburg NBE, Cidlowski JA (1995) The role of DNA fragmentation in apoptosis. Trends Cell BioI 5:21-26
Collins RJ, Harmon BV, Gobe GC, Kerr JFR (1992) Internucleosomal DNA
cleavage should not be the sole criterion for identifying apoptosis. Int J
Radiat Res 61:451-453
Dawson BA, Lough J (1988) Immunocytochemical localization of transient
DNA strand breaks in differentiating myotubes using in situ nick-translation. Dev BioI 127:362-367
Farzaneh F, Zalin R, Brill D, Shall S (1982) DNA strand breaks and ADPribosyl transferase activity in eukaryotic differentiation. Nature 300:362366
Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed
cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell
BioI 119:493-501
Gold R, Schmied M, Breitschopf H, Hartung HP, Toyka KV, Lassmann H
(1994) Differentiation between cellular apoptosis and necrosis by the