Current Concepts in Cell Death

Boris Zhivotovsky1, Sten Orrenius1

1 Karolinska Institutet, Stockholm, Sweden
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 18.1
DOI:  10.1002/0471143030.cb1801s11
Online Posting Date:  August, 2001
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

This unit attempts to define cell toxicity that leads to cellular demise, with a strong emphasis on cell death via both apoptosis and necrosis, by summarizing some of the more recent developments in cellular, molecular, and biochemical studies of the events that govern the induction and execution of cell death. Specific topics pertaining to cell death include structural changes, macromolecular degradation, cellular signaling, the role of mitochondria, and genetic modulation of apoptotic cell death.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Cell Death
  • Structural Changes During Cell Death
  • Macromolecular Degradation During Cell Death
  • Cellular Signaling During Cell Death
  • Role of Mitochondria During Cell Death
  • Genetic Modulation of Apoptotic Cell Death
  • Conclusions
  • Literature Cited
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Alnemri, E.S., Livingston, D.J., Nicholson, D.W., Salvesen, G., Thornberry, N.A., Wong, W.W., and Yuan, J. 1996. Human ICE/CED‐3 protease nomenclature. Cell 87:171.
   Ankarcrona, M., Dypbukt, J.M., Bonfoco, E., Zhivotovsky, B., Orrenius, S., Lipton, S.A., and Nicotera, P. 1995. Glutamate‐induced neuronal death:A succession of necrosis or apoptosis depending on mitochondrial function. Neuron 15:961‐973.
   Arai, A., Kessler, M., Lee, K., and Lynch, G. 1990. Calpain inhibitors improve the recovery of synaptic transmission from hypoxia in hippocampal slices. Brain Res. 532:63‐68.
   Arends, M.J., McGregor, A.H., Toft, N.J., Brown, E.J.H., and Wyllie, A.H. 1993. Susceptibility to apoptosis is differentially regulated by c‐myc and Ha‐ras oncogenes and is associated with endo‐nuclease availability. Br. J. Cancer 68:1127‐1133.
   Armstrong, R.C., Aja, T.J., Hoang, K.D., Gaur, S., Bai, X., Alnemri, E., Litwack, G., Karanewsky, D.S., Fritz, L.C., and Tomaselli, K.J. 1997. Activation of the CED‐3/ICE‐related protease CPP32 in cerebellar granule neurons undergoing apoptosis, but not necrosis. J. Neurosci. 17:553‐562.
   Arora, A.S., de Groen, P.C., Croal, D.E., Emori, Y., and Gores, G.J. 1996. Hepatocellular carcinoma cells resist necrosis during anoxia by preventing phospholipase‐mediated calpain activation. J. Cell. Physiol. 167:434‐442.
   Bakhshi, A., Jensen, J.P., Goldman, P., Wright, J.J., McBride, O.W., Epstein, A.L., and Korsmeyer, S.J. 1985. Cloning the chromosomal breakpoint of t(14;18) human lymphomas:Clustering around JH on chromosome 14 and near a transcriptional unit on 18. Cell 41:899‐906.
   Baxter, G.D. and Lavin, M.F. 1992. Specific protein phosphorylation in apoptosis induced by ionizing radiation and heat shock in human lymphoid tumor lines. J. Immunol. 148:1949‐1954.
   Bernelli‐Zazzera, A. 1975. Ribosomes in dying liver cells. In Pathogenesis and Mechanisms of Liver Cell Necrosis (D. Keppler, ed.)pp. 103‐111. MTP Press, Lancaster.
   Boe, R., Gjertsen, B.T., Vintermyr, O.K., Houge, G., Lanotte, M., and Doskeland, S.O. 1991. The protein phosphatase inhibitor okadaic acid induces morphological changes typical of apoptosis in mammalian cells. Exp. Cell Res. 195:237‐246.
   Bowen, I.D. 1981. Techniques for demonstrating cell death. In Cell Death in Biology and Physiology (I.D. Bowen and R.A. Lockshin, eds.) pp. 379‐444. Chapman & Hall, New York.
   Brown, D.G., Sun, X.M., and Cohen, G.M. 1993. Dexamethasone‐induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation. J. Biol. Chem. 268:3037‐3039.
   Budihardjo, I., Oliver, H., Lutter, M., Luo, X., and Wang, X. 1999. Biochemical pathways of caspase activation during apoptosis. Ann. Rev. Cell Dev. Biol. 15:269‐290.
   Croal, D.E. and DeMartino, G.N. 1991. Calcium‐activated neutral protease (calpain) system:Structure, function and regulation. Physiol. Rev. 71:813‐847.
   Daugas, E., Susin, S.A., Zamzami, N., Ferri, K.F., Irinopoulou, T., Larochette, N., Prevost, M.C., Leber, B., Andrews, D., Penninger, J., and Kroemer, G. 2000. Mitochondrio‐nuclear translocation of AIF in apoptosis and necrosis. FASEB J. 14:729‐739.
   de Duve, C. 1963. The lysosome concept. In Lysosomes (A.V.S. deReuck and M.P. Cameron, eds.) pp. 101‐118. Churchill Livingston, London.
   Dickinson, J.L., Bates, E.J., Ferrante, A., and Antalis, T.M. 1995. Plasminogen activator inhibitor type 2 inhibits tumor necrosis factor alpha–induced apoptosis. Evidence for an alternate biological function. J. Biol. Chem. 270:27894‐27904.
   Duvall, E., Wyllie, A.H., and Morris, R.G. 1985. Macrophage recognition of cells undergoing cell death (apoptosis). Immunology 56:351‐358.
   Evan, G. and Littlewood, T. 1998. A matter of life and cell death. Science 281:1317‐1322.
   Evan, G.I., Wyllie, A.H., Gilbert, C.S., Littlewood, T.D., Hartmut, L., Brooks, M., Waters, C.M., Penn, L.Z., and Hancock, D. 1992. Induction of apoptosis in fibroblasts by c‐myc protein. Cell 69:119‐128.
   Fadeel, B., Zhivotovsky, B., and Orrenius, S. 1999. All along the watchtower:On the regulation of apoptosis regulators. FASEB J. 13:1647‐1657.
   Fadeel, B., Orrenius, S., and Zhivotovsky, B. 2000. The most unkindest cut of all:On the multiple roles of mammalian caspases. Leukemia 14:1514‐1525.
   Fadok, V., Bratton, D.L., Rose, D.M., Pearson, A., Ezekewitz, R.A., and Henson, P.M. 2000. A receptor for phosphatidylserine‐specific clearance of apoptotic cells. Nature 405:85‐90.
   Ferri, K.F. and Kroemer, G. 2001. Mitochondria–the suicide organelles. BioEssays 23:111‐115.
   Fesus, L., Thomazy, V., and Falus, A. 1987. Induction and activation of tissue transglutaminase during programmed cell death. FEBS Lett. 224:104‐108.
   Forbes, I.J., Zalewski, P.D., Gianniakis, C., and Cowled, P.A. 1992. Induction of apoptosis in chronic lymphocytic leukaemia cells and its prevention by phorbol esters. Exp. Cell Res. 198:367‐372.
   Glucksmann, A. 1951. Cell death is normal vertebrate ontology. Biol. Rev. 26:59‐86.
   Gorczyca, W., Bruno, S., Darzynkiewicz, R.J., Gong, J., and Darzynkiewicz, Z. 1992. DNA strand breaks occurring during apoptosis:Their early detection by terminal deoxynucleotide transferase and nick translation assays and prevention by serine protease inhibitors. Int. J. Oncol. 1:639‐648.
   Gross, A., McDonel, J.M., and Korsmeyer, S.J. 1999. Bcl‐2 family members and the mitochondria in apoptosis. Genes Dev. 13:1899‐1911.
   Haslett, C. 1992. Resolution of acute inflammation and the role of apoptosis in tissue fate of granulocytes. Clin. Sci. 83:639‐648.
   Hengartner, M.O. 2000. The biochemistry of apoptosis. Nature 407:770‐776.
   Hockenbery, D.M., Nunez, G., Milliman, C., Schreiber, R.D., and Korsmeyer, S.J. 1990. Bcl‐2 in inner mitochondrial membrane protein that blocks programmed cell death. Nature 348:334‐336.
   Hockenbery, D.M., Oltvai, Z.N., Yin, X.M., Milliman, C.L., and Korsmeyer, S.J. 1993. Bcl‐2 functions in an antioxidant pathway to prevent apoptosis. Cell 75:241‐251.
   Holownia, A., Ledig, M., and Menez, J.F. 1997. Ethanol‐induced cell death in cultured rat astroglia. Neurotoxicol. Teratol. 19:141‐146.
   Houge, G. and Doskeland, S.O. 1996. Divergence towards a dead end? Cleavage of the divergent domain of ribosomal RNA in apoptosis. Experientia 52:963‐967.
   Houge, G., Doskeland, S.O., Boe, R., and Lanotte, M. 1993. Selective cleavage of 28S rRNA variable regions V3 and V13 in myeloid leukemia cell apoptosis. FEBS Lett. 315:16‐20.
   Jaattela, M., Wissing, D., Bauer, P., and Li, G.C. 1992. Major heat shock protein hsp70 protects tumor cells from tumor necrosis factor cytotoxicity. EMBO J. 11:3507‐3512.
   Jacobson, M.D., Burne, J.F., King, M.P., Miyashita, T., Reed, J.C., and Raff, M.C. 1993. Bcl‐2 blocks apoptosis in cells lacking mitochondrial DNA. Nature 361:365‐369.
   Jensen, P.H., Cressey, L.I., Gjertsen, B.T., Madsen, P., Mellgren, G., Hokland, P., Gliemann, J., Doskeland, S.O., Lanotte, M., and Vintermyr, O.K. 1994. Cleaved intracellular plasminogen activator inhibitor 2 in human myeloleukaemia cells is a marker of apoptosis. Br. J. Cancer 70:834‐840.
   Johnson, D.E. 2000. Noncaspase proteases in apoptosis. Leukemia 14:1695‐1703.
   Kaiser, N. and Edelman, I.S. 1978. Further studies on the role of calcium in glutocorticoid‐induced lympholysis. Endocrinology 103:936‐942.
   Kass, G.E.N., Eriksson, J.E., Weis, M., Orrenius, S., and Chow, S.C. 1996. Chromatin condensation during apoptosis requires ATP. Biochem. J. 318:749‐752.
   Kastan, M.B., Onyekwere, O., Sidransky, D., Vogelstein, B., and Craig, R.W. 1991. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 51:6304‐6311.
   Kerr, J.F.R. 1971. Shrinkage necrosis:A distinct mode of cellular death. J. Pathol. 105:13‐20.
   Kerr, J.F.R., Wyllie, A.H., and Currie, A.R. 1972. Apoptosis: A basic biological phenomenon with wide ranging implications in tissue kinetics. Br. J. Cancer 26:239‐257.
   Kizaki, H., Tadakuma, T., Odaka, C., Maramatsu, J., and Isimura, Y. 1989. Activation of suicide process of thymocytes through DNA fragmentation by calcium ionophores and phorbol esters. J. Immunol. 143:1790‐1794.
   Koopman, G., Reutelingsperger, C.P.M., Kuijten, G.A.M., Keehnen, R.M.J., Pals, S.T., and van Oers, M.H.J. 1994. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415‐1420.
   Krajewski, S., Krajewska, M., Shabaik, A., and Reed, J.C. 1994. Immunohistochemical determination of in vivo distribution of bax, a dominant inhibitor of bcl‐2. Am. J. Pathol. 145:1323‐1336.
   Krammer, P.H. 2000. CD95's deadly mission in the immune system. Nature 407:789‐795.
   Kroemer, G. 1997. Mitochondrial implication in apoptosis: Towards an endosymbiont hypothesis of apoptosis evolution. Cell Death Differ. 4:443‐456.
   Lavin, M.F., Watters, D., and Song, Q. 1996. Role of protein kinase activity in apoptosis. Experientia 52:979‐994.
   Lazebnik, Y.A., Cole, S., Cooke, C.A., Nelson, W.G., and Earnshaw, W.C. 1993. Nuclear events of apoptosis in vitro in cell‐free mitotic extracts: A model system for analysis of the active phase of apoptosis. J. Cell Biol. 123:7‐22.
   De Levrero, M., Laurenzi, V., Costanzo, A., Sabatini, S., Gong, J., Wang, J.Y.J., and Melino, G. 2000. The p53/p63/p73 family of transcription factors: Overlapping and distinct functions. J. Cell Sci. 113:1661‐1670.
   Lezoulac'h, F., Rupprecht, R., Holsboer, F., and Behl, C. 1996. Bcl‐2 prevents hippocampal cell death induced by the neuroleptic drug haloperidol. Brain Res. 738:176‐179.
   Lindquist, S. and Craig, E.A. 1988. The heat shock proteins. Annu. Rev. Genet. 22:631‐677.
   Liu, X., Kim, N.C., Yang, J., Jemmerson, R., and Wang, X. 1996. Induction of apoptotic program in cell‐free extracts: Requirement for dATP and cytochrome c. Cell 86:147‐157.
   Liu, X., Zou, H., Widlak, P., Garrard, W., and Wang, X. 1999. Activation of the apoptotic endonuclease DFF40 (caspase‐activated DNase or nuclease). Oligomerization with histone H1. J. Biol. Chem. 274:13836‐13840.
   Lockshin, R.A. 1981. Cell death in metamorphosis. In Cell Death in Biology and Physiology (I.D. Bowen and R.A. Lockshin, eds.) pp. 79‐121. Chapman & Hall, New York.
   Lowe, S.W., Schmitt, E.M., Smith, S.W., Osborne, B.A., and Jacks, T. 1993. p53 is required for radiation‐induced apoptosis in mouse thymocytes. Nature 362:847‐849.
   Mailhos, C., Howard, M.K., and Latchman, D.S. 1993. Heat shock protects neuronal cells from programmed cell death by apoptosis. Neuroscience 55:621‐627.
   Martin, S.J., Lennon, S.V., Bonham, A.M., and Cotter, A.M. 1990. Induction of apoptosis (programmed cell death) in human leukemic HL‐60 cells by inhibition of RNA or protein synthesis. J. Immunol. 145:1859‐1867.
   McClean, S. and Hill, B.T. 1992. An overview of membrane, cytosolic and nuclear proteins associated with the expression of resistance to multiple drugs in vitro. Biochem. Biophys. Acta 1114:107‐127.
   McConkey, D.J., Zhivotovsky, B., and Orrenius, S. 1996. Apoptosis: Molecular mechanisms and biomedical implications. Mol. Aspects Med. 17:1‐110.
   McIlroy, D., Tanaka, M., Sakahira, H., Fukuyama, H., Suzuki, M., Yamamura, K.‐I., Uchiyama, Y., and Nagata, S. 2000. An auxiliary mode of apoptotic DNA fragmentation provided by phagocytes. Genes Dev. 14:549‐558.
   Mirabelli, F., Salis, A., Vairetti, M., Bellomo, G., Thor, H., and Orrenius, S. 1989. Cytoskeletal alterations in human platelets exposed to oxidative stress are mediated by oxidative and Ca2+‐dependent mechanisms. Arch. Biochem. Biophys. 270:478‐488.
   Mosser, D.D. and Martin, L.H. 1992. Induced thermotolerance to apoptosis in a human T lymphocyte cell line. J. Cell. Physiol. 151:561‐570.
   Nagata, S. 2000. Apoptotic DNA fragmentation. Exp. Cell Res. 256:12‐18.
   Newmeyer, D.D., Farston, D.M., and Reed, J.C. 1994. Cell‐free apoptosis in Xenopus egg extracts: Inhibition by Bcl‐2 and requirement for an organelle fraction enriched in mitochondria. Cell 79:353‐364.
   Nicholson, D.W. and Thornberry, N.A. 1997. Caspases: Killer proteases. Trends Biochem. Sci. 22:299‐306.
   Nicotera, P. and Leist, M. 1997. Energy supply and the shape of cell death in neurones and lymphoid cells. Cell Death Differ. 4:435‐442.
   Nicotera, P. and Orrenius, S. 1998. The role of calcium in apoptosis. Cell Calcium 23:173‐180.
   Nicotera, P., Hartzell, P., Davis, G., and Orrenius, S. 1986. The formation of plasma membrane blebs in hepatocytes exposed to agents that increased cytosolic Ca2+ is mediated by the activation of a non‐lysosomal proteolytic system. FEBS Lett. 209:139‐144.
   Nishizuka, Y. 1984. The role of PKC in cell surface signal transduction and tumour promotion. Nature 308:693‐698.
   Nobel, S. 1997. Thiol redox state in apoptosis: Physiological and toxicant modulation. Ph.D. Thesis, Karolinska Institutet, Stockholm.
   Oesterreich, S., Weng, C., Qiu, M., Hilsenbeck, S.G., Osborne, C.K., and Fuqua, S.A.W. 1993. The small heat shock protein hsp27 is correlated with growth and drug resistance in human breast cancer cell lines. Cancer Res. 53:4443‐4448.
   Orlowski, R. Z. 1999. The role of the ubiquitin‐proteasome pathway in apoptosis. Cell Death Differ. 6:303‐313.
   Robertson, J.D., Orrenius, S., and Zhivotovsky, B. 2000. Nuclear events in apoptosis. J. Struct. Biol. 129:346‐358.
   Ryan, K.M. and Birnie, G.D. 1996. Myc oncogenes: The enigmatic family. Biochem. J. 314:713‐721.
   Salvesen, G.S. 2001. A lysosomal protease enters the death scene. J. Clin. Invest. 107:21‐22.
   Samali, A. and Cotter, T.G. 1996. Heat shock proteins increase resistance to apoptosis. Exp. Cell Res. 223:163‐170.
   Samali, A., Gilje, B., Doskeland, S.O., Cotter, T.G., and Houge, G. 1997. The ability to cleave 28S rRNA during apoptosis is a cell type dependent trait unrelated to DNA fragmentation. Cell Death Differ. 4:289‐293.
   Samali, A., Nordgren, H., Zhivotovsky, B., Peterson, E. and Orrenius, S. 1999a. A comparative study of apoptosis and necrosis in HepG2 cells. Oxidant‐induced caspase activation leads to necrosis. Biochem. Biophys. Res. Commun. 256:6‐11.
   Samali, A., Cai, J., Zhivotovsky, B., Jones, D.P., and Orrenius, S. 1999b. Presence of a pre‐apoptotic complex of procaspase‐3, Hsp60 and Hsp10 in the mitochondria fraction of Jurkat cells. EMBO J. 18:2040‐2048.
   Saunderson, C.J. 1982. Morphological aspects of lymphocyte mediated cytotoxicity. In Mechanisms of Cell Mediated Cytotoxicity (W.R. Clark and P. Golstein, eds.) pp. 3‐21. Plenum, New York.
   Savill, J. and Fadok, V. 2000. Corpse clearance defines the meaning of cell death. Nature 407:784‐788.
   Shi, L., Kam, C.M., Powers, J.C., Aebersold, R., and Greenberg, A. 1992. Purification of three cytotoxic lymphocyte granule serine proteases that induce apoptosis through distinct substrate and target cell interaction. J. Exp. Med. 176:1521‐1529.
   Shimizu, S., Eguchi, Y., Kamiike, W., Itoh, Y., Hasegawa, J., Yamabe, K., Otsuki, Y., Matsuda, H., and Tsujimoto, Y. 1996. Induction of apoptosis as well as necrosis by hypoxia and predominant prevention of apoptosis by Bcl‐2 and Bcl‐xL. Cancer Res. 56:2161‐2166.
   Shinohara, K., Tomioka, M., Nakano, H., Tone, S., Ito, H., and Kawashima, S. 1996. Apoptosis induction resulting from proteasome inhibition. Biochem. J. 317:385‐388.
   Sims, K.D. and Robinson, M.B. 1999. Expression patterns and regulation of glutamate transporters in the developing and adult nervous system. Crit. Rev. Neurobiol. 13:169‐197.
   Somasundaram, K. and El‐Deiry, W.S. 2000. Tumor suppressor p53: Regulation and function. Frontiers Biosci. 5:D424‐D437.
   Sperandio, S., de Belle, I., and Bredesen, D.E. 2000. An alternative, non‐apoptotic form of programmed cell death. Proc. Natl. Acad. Sci. U.S.A. 97:14376‐14381.
   Stennicke, H.R. and Salvesen, G.S. 2000. Caspases–controlling intracellular signals by protease zymogen activation. Biochim. Biophys. Acta 1477:299‐306.
   Stoka, V., Turk, B., Schendel, S.L., Kim, T.‐H., Cirman, T., Snipas, S.J., Ellerby, L.M., Bredesen, D., Freeze, H., Abrahamson, M., Brömme, D., Krajewski, S., Reed, J.C., Yin, X.‐M., Turk, V., and Salvesen, G.S. 2001. Lysosomal protease pathways to apoptosis. Cleavage of Bid, not procaspases, is the most likely route. J. Biol. Chem. 276:3149‐3157.
   Sun, X., Shimizu, H., and Yamamoto, K. 1995. Identification of a novel p53 promoter element involved in genotoxic stress‐inducible p53 gene expression. Mol. Cell. Biol. 15:4489‐4496.
   Trump, B.F. and Berezesky, I.K. 1995. Calcium‐mediated cell injury and cell death. FASEB J. 9:219‐228.
   Trump, B.F. and Ginn, F.L. 1969. The pathogenesis of subcellular reaction to lethal injury. In Methods and Achievements in Experimental Pathology, vol. 4 (E. Bajusz and G. Jasmin, eds.) p. 1. Karger, Basel.
   Tsujimoto, Y. 1997. Apoptosis and necrosis: Intracellular ATP level as a determinant for cell death modes. Cell Death Differ. 4:429‐434.
   Vanags, D.M., Pörn‐Ares, M.A., Coppola, S., Burgess, D.H., and Orrenius, S. 1996. Protease involvement in fodrin cleavage and phosphatidylserine exposure in apoptosis. J. Biol. Chem. 271:31075‐31085.
   Van Lancker, J.L. 1975. Hydrolases and cellular death. In Pathogenesis and Mechanisms of Liver Cell Necrosis (D. Keppler, ed.) pp. 25‐35. MTP Press, Lancaster.
   Wolf, C.M. and Eastman, A. 1999. Intracellular acidification during apoptosis can occur in the absence of a nucleus. Biochem. Biophys. Res. Commun. 254:821‐827.
   Wu, Y.‐C., Stanfiels, G.M., and Horvitz, H.R. 2000. Nuc‐1, a Caenorhabditis elegans DNase II homolog, functions in an intermediate step of DNA degradation during apoptosis. Genes Dev. 14:536‐548.
   Wyllie, A.H. 1980. Glutocorticoid‐induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555‐556.
   Wyllie, A. and Golstein, P. 2001. More than one way to go. Proc. Natl. Acad. Sci. U.S.A. 98:11‐13.
   Wyllie, A.H., Rose, K.A., Morris, R.G., Steel, C.M., Forster, E., and Spandidos, D.A. 1987. Rodent fibroblast tumors expressing human myc and ras genes: Growth, metastasis and endogenous oncogene expression. Br. J. Cancer 56:251‐259.
   Xanthoudakis, S. and Nicholson, D.W. 2000. Heat‐shock proteins as death determinants. Nature Cell Biol. 2:E163‐E165.
   Yang, A. and McKeon, F. 2000. p63 and p73: p53 mimics, menaces and more. Nature Rev. Mol. Cell Biol. 1:199‐207.
   Yasuhara, N., Eguchi, Y.M., Tachibana, T., Imamoto, N., Yoneda, Y., and Tsujimoto, Y. 1997. Essential role of active nuclear transport in apoptosis. Genes Cells 2:55‐64.
   Yonish‐Rouach, E., Resnitzky, D., Lotem, J., Sachs, L., Kim‐Chi, A., and Oren, M. 1991. Wild‐type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interlukin‐6. Nature 353:345‐347.
   Yu, S.P. and Choi, D.W. 2000. Ions, cell volume, and apoptosis. Proc. Natl. Acad. Sci. U.S.A. 97:9360‐9362.
   Yuan, J., Shaham, S., Ledoux, S., Ellis, H.M., and Horwitz, R.H. 1993. The C. elegans cell death gene ced‐3 encodes a protein similar to the mammalian interleukin‐1β‐converting enzyme. Cell 75:641‐652.
   Zamzami, N., Marchetti, P., Castedo, M., Decaudin, D., Macho, A., Hirsch, T., Susin, S.‐A., Petit, P.X., Mignotte, B., and Kroemer, G. 1995. Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death. J. Exp. Med. 182:367‐377.
   Zhivotovsky, B., Wade, D., Nicotera, P., and Orrenius, S. 1994. Role of nucleases in apoptosis. Int. Arch. Allergy Immunol. 105:333‐338.
   Zhivotovsky, B., Burgess, D.H., Vanags, D.M., and Orrenius, S. 1997. Involvement of cellular proteolytic machinery in apoptosis. Biochem. Biophys. Res. Commun. 230:481‐488.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library