Abstract
Apoptosis is an active process of selective cell death that occurs during development and has been implicated in the pathogenesis of a variety of human diseases (Thompson, 1995). Apoptosis can be distinguished from the other major form of cell death, necrosis, on the basis of both morphological and biochemical features; these include chromatin condensation with nuclear pyknosis, and cytoplasmic shrinkage (Thompson, 1995). One prominent hallmark of apoptosis is a characteristic pattern of DNA cleavage into oligonucleosome-sized fragments that, under most circumstances, can be visualized by agarose gel electrophoresis. However, when relatively few apoptotic cells are present, fragmented DNA can be labeled in situ by more sensitive techniques. Using this approach, apoptotic cell death has been documented in the rodent central nervous system (CNS) following cerebral ischemia (Heron et al., 1993; MacManus et al, 1994), status epilepticus (Filipowski et al., 1994; Pollard et al., 1994; Sakhi et al., 1994) and adrenalectomy (Schreiber et al., 1994). Similar methods have been used to demonstrate apoptotic cells in the brain of patients with Alzheimer’s (Su et al., 1994; Lassamn et al., 1995) and Huntington’s diseases (Portera-Cailliau et al., 1995).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Cohen J. J. (1993) Apoptosis Immunol Today 14, 126–130.
Colotta F, Polentarutti N., Sironi M., and Mantovani A (1992) Expression and involvement of c-fos and c-jun protooncogenes in programmed cell death induced by growth factor deprivation in lymphoid cell lines J Biol Chem 267, 18,278–18,283.
Crumrine R C, Thomas A L, and Morgan P. F. (1994) Attenuation of p53 expression protects against focal ischemic damage in transgenic mice J. Cereb Blood Flow Metab 14, 887–891
Donehower L A., Harvey M, Slagle B. L., McArthur M. J, Montgomery C. A. Jr, Butel J. S., and Bradley A. (1992) Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature 356, 215–221.
Dragunow M, Beilharz E, Sirimanne E., Lawlor P, Williams C, Bravo R., and Gluckman P (1994) Immediate-early gene protein expression in neurons undergoing delayed death but not necrosis following hypoxic-ischemic injury to the young rat brain. Mol Brain Res 25, 19–33.
El-Deiry W. S., Harper J W., O’Connor P M, Velculescu V E, Canman C. E., Jackman J, Pietenpol J A., Burrell M., Hill D E, Wang Y., Wiman K. G, Mercer W. E., Kastan M B., Kohn K. W., Elledge S J, Kinzler K W, and Vogelstein B. (1994) WAF1/CIP1 is induced in p53-mediated Gl arrest and apoptosis. Cancer Res 54, 1169–1174
Farmer G., Bargonetti J, Zhu H, Friedman P, Prywes R., and Prives C (1992) Wild-type p53 activates transcription in vitro Nature 358, 83–86.
Filipowski R K, Hetman M, Kaminska B, and Kaczmarek L. (1994) DNA fragmentation in rat brain after intraperitoneal administration of kainate NeuroReport 5, 1538–1540.
Freeman R S., Estus S., and Johnson E M (1994) Analysis of cell cycle-related gene expression in postmitotic neurons: selective induction of cyclin D1 during programmed cell death. Neuron 12, 343–355
Ham J., Babij C, Whitfield J, Pfarr C. M., Lallemand D., Yaniv M., and Rubin L. L (1995) A c-Jun dominant negative mutant protects sympathetic neurons against programmed cell death Neuron 14, 927–939
Heron A, Pollard H., Dessi F, Moreau J, Lasbennes F, Ben-Ari Y., and Charriaut-Marlangue C (1993) Regional variability in DNA fragmentation after global ischemia evidenced by combined histological and gel electrophoresis observations in the rat brain J Neurochem 61, 1973–1976
Hollstein M, Sidransky D, Vogelstein B., and Harris C C. (1991) p53 mutations in human cancers Science 253, 49–53
Kastan M B, Onyerkwere O., Sidransky D, Vogelstein B, and Craig R W (1991) Participation of p53 protein in the cellular response to DNA damage Cancer Res 53, 6304–6311
Kern S E., Kinzler K. W, Bruskin A, Jarosz D, Friedman P., Prives C, and Vogelstein B. (1991) Identification of p53 as a sequence-specific DNA-binding protein Science 252, 1708–1711.
Kuerbitz S J, Plunkett B S., Walsh W. V, and Kastan M. B. (1992) Wild-type p53 is a cell cycle checkpoint determinant following irradiation Proc Natl Acad Sci USA 89, 7491–7495.
Lane D. P. (1992) p53, guardian of the genome Nature 358, 15–16.
Lassman H., Bancher C, Breitschopf H., Wegiel J., Bobmski M., Jellinger K., and Wismewski H M (1995) Cell death in Alzheimer’s disease evaluated by DNA fragmentation in situ Acta Neuropathol 89, 35–41
Levine A. J., Momand J, and Finlay C. A. (1991) The p53 tumor suppressor gene. Nature 351, 453–456
Li Y., Chopp M., Zhang Z. G., Zaloga C, Niewenhuis L., and Gautam S. (1994) p53-immunoreactive protein and p53 mRNA expression after transient middle cerebral artery occlusion in rats. Stroke 25, 849–856
MacManus J P, Hill I E, Huang Z-G., Rasquinha I., Xue D, and Buchan A. M. (1994) DNA damage consistent with apoptosis in transient focal ischaemic neocortex. NeuroReport 5, 493–496.
Manev H., Kharlamov A, and Armstrong D. M (1994) Photochemical brain injury in rats triggers DNA fragmentation p53 and HSP72 NeuroReport 5, 2661–2664.
Nitecka L, Tremblay E., Charton G, Bouillot J. P., Berger M. L, and Ben-Ari Y (1984) Maturation of kainic acid seizure-brain damage syndrome in the rat II Histo-pathological sequelae Neuroscience 13, 1073–1094
Pollard H., Charriaut-Marlangue C, Cantagrel S, Represa A, Robain O., Moreau J., and Ben-Ari Y (1994) Kainate-induced apoptotic cell death in hippocampal neurons Neuroscience 63, 7–18
Portera-Cailliau C, Hedreen J C, Price D L., and Koliatsos V. E. (1995) Evidence for apoptotic cell death in Huntington disease and excitotoxic animal models J Neurosci. 15, 3775–3787
Ryan J. J., Danish R., Gottlieb C A., and Clarke M. F (1993) Cell cycle analysis of p53-mduced death in murine erythroleukemia cells Mol Cell Biol 13, 711–719.
Sakhi S, Bruce A., Sun N, Tocco G, Baudry M, and Schreiber S S (1994) p53 induction is associated with neuronal damage in the central nervous system Proc Natl Acad Sci USA 91, 7525–7529.
Sakhi S, Bruce A., Sun N, Tocco G., Baudry M, and Schreiber S. S. (1995) Excitotoxin-mduced apoptosis in organotypic hippocampal cultures p53 upregulation and DNA fragmentation Exp Neurol, in press.
Schreiber S. S., Sakhi S., Millicent M. Dugich-Djordjevic and Nichols N. R (1994) Tumor suppressor p53 induction and DNA damage in hippocampal granule cells after adrenalectomy. Exp. Neurol 130, 368–376.
Schreiber S. S, Tocco G., Najm I, Thompson R. F., and Baudry M. (1993) Cycloheximide prevents kainate-induced neuronal death and c-fos expression in adult rat brain J Mol Neurosci 4, 149–159.
Schwob J. E., Fuller T., Price J. L, and Olney J. W. (1980) Widespread patterns of neuronal damage following systemic or intracerebral injections of kainic acid: a histological study. Neuroscience 5, 991–1014
Shaw P., Bovey R, Tardy S., Sahli R, Sordat B, and Costa J. (1992) Induction of apoptosis by wild-type p53 in a human colon tumor-derived cell line. Proc Natl Acad Sci USA 89, 4495–4499
Shi Y., Glynn J M., Guilbert L J., Cotter T. G., Bissonnette R P., and Green D R. (1992) Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas Science 257, 212–214
Sloviter R S, Dean E, and Neubort S. (1993a) Electron microscopic analysis of adrenalectomy-induced hippocampal granule cell degeneration in the rat: apoptosis in the adult central nervous system J Comp Neurol 330, 337–351
Sloviter R. S., Sollas A L, Dean E., and Neubort S (1993b) Adrenalectomy-induced granule cell degeneration in the rat hippocampal dentate gyrus: characterization of an in vivo model of controlled neuronal death J Comp Neurol 330, 324–336
Smeyne R. J., Vendrell M, Hayward M., Baker S J, Miao G. G, Schilling K., Robertson L. M., Curran T., and Morgan J I. (1993) Continuous c-fos expression precedes programmed cell death in vivo Nature 363, 166–169
Steller H. (1995) Mechanisms and genes of cellular suicide Science 267, 1445–1449.
Su J H., Anderson A J, Cummings B J, and Cotman C. W. (1994) Immunohistochemical evidence for apoptosis in Alzheimer’s disease NeuroReport 5, 2529–2533
Thompson C B (1995) Apoptosis in the pathogenesis and treatment of disease Science 267, 1456–1462.
Wood K. A and Youle R J. (1995) The role of free radicals and p53 in neuron apoptosis in vivo, J Neurosci 15, 5851–5857.
Yonish-Rouach E, Resnitzky D, Lotem J, Sachs L, Kimchi A, and Oren M. (1991) Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6. Nature 352, 345–347
Zhan Q., Carrier F., and Fornace A. J., Jr (1993) Induction of cellular p53 activity by DNA-damaging agents and growth arrest Mol Cell Biol 13, 4242–4250.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1997 Humana Press Inc.
About this protocol
Cite this protocol
Sakhi, S., Schreiber, S.S. (1997). p53 Induction Is a Marker of Neuronal Apoptosis in the Central Nervous System. In: Poirier, J. (eds) Apoptosis Techniques and Protocols. Neuromethods, vol 29. Humana Press. https://doi.org/10.1385/0-89603-451-8:85
Download citation
DOI: https://doi.org/10.1385/0-89603-451-8:85
Publisher Name: Humana Press
Print ISBN: 978-0-89603-451-8
Online ISBN: 978-1-59259-634-8
eBook Packages: Springer Protocols