Activation of CPP32-Like Caspases Contributes to Neuronal Apoptosis and Neurological Dysfunction after Traumatic Brain Injury

We examined the temporal profile of apoptosis after fluid percussion-induced traumatic brain injury (TBI) in rats and investigated the potential pathophysiological role of caspase-3-like proteases in this process. DNA fragmentation was observed in samples from injured cortex and hippocampus, but not from contralateral tissue, beginning 4 hr after TBI and continuing for at least 3 d. Double labeling of brain with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and an antibody directed to neuronal nuclear protein identified apoptotic neurons with high frequency in both traumatized rat cortex and hippocampus. Cytosolic extracts from injured cortex and hippocampus, but not from contralateral or control tissue, induced internucleosomal DNA fragmentation in isolated nuclei with temporal profiles consistent with those of DNA fragmentation observed in vivo. Caspase-3 mRNA levels, estimated by semiquantitative RT-PCR, were elevated fivefold in ipsilateral cortex and twofold in hippocampus by 24 hr after TBI. Caspase-1 mRNA content also was increased after trauma, but to a lesser extent in cortex. Increased caspase-3-like, but not caspase-1-like, enzymatic activity was found in cytosolic extracts from injured cortex. Intracerebroventricular administration of z-DEVD-fmk—a specific tetrapeptide inhibitor of caspase-3—before and after injury markedly reduced post-traumatic apoptosis, as demonstrated by DNA electrophoresis and TUNEL staining, and significantly improved neurological recovery. Together, these results implicate caspase-3-like proteases in neuronal apoptosis induced by TBI and suggest that the blockade of such caspases can reduce post-traumatic apoptosis and associated neurological dysfunction.

[1]  A. Yakovlev,et al.  The Role of CED-3-Related Cysteine Proteases in Apoptosis of Cerebellar Granule Cells , 1997, The Journal of Neuroscience.

[2]  Xiaodong Wang,et al.  DFF, a Heterodimeric Protein That Functions Downstream of Caspase-3 to Trigger DNA Fragmentation during Apoptosis , 1997, Cell.

[3]  M. Moskowitz,et al.  Inhibition of interleukin 1beta converting enzyme family proteases reduces ischemic and excitotoxic neuronal damage. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[4]  G. Salvesen,et al.  FLICE Induced Apoptosis in a Cell-free System , 1997, The Journal of Biological Chemistry.

[5]  T. Miller,et al.  Metabolic and Genetic Analyses of Apoptosis in Potassium/Serum-Deprived Rat Cerebellar Granule Cells , 1996, The Journal of Neuroscience.

[6]  Junying Yuan,et al.  Human ICE/CED-3 Protease Nomenclature , 1996, Cell.

[7]  A. Yakovlev,et al.  A new approach for the electrophoretic detection of apoptosis. , 1996, Nucleic acids research.

[8]  H. Manev,et al.  Pharmacological characterization of apoptotic cell death in a model of photothrombotic brain injury in rats , 1996, Brain Research.

[9]  D. Bredesen,et al.  Bcl-2 Expression in Neural Cells Blocks Activation of ICE/CED-3 Family Proteases during Apoptosis , 1996, The Journal of Neuroscience.

[10]  J. Gybels,et al.  Production of tumor necrosis factor in spinal cord following traumatic injury in rats , 1996, Journal of Neuroimmunology.

[11]  M. Weller,et al.  Potassium Deprivation-Induced Apoptosis of Cerebellar Granule Neurons: A Sequential Requirement for New mRNA and Protein Synthesis, ICE-Like Protease Activity, and Reactive Oxygen Species , 1996, The Journal of Neuroscience.

[12]  S. Srinivasula,et al.  In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[13]  T. Calcaterra,et al.  Regulation of Transforming Growth Factor‐β1‐Mediated Apoptosis in Head and Neck Squamous Cell Carcinoma , 1996, The Laryngoscope.

[14]  David Wallach,et al.  Involvement of MACH, a Novel MORT1/FADD-Interacting Protease, in Fas/APO-1- and TNF Receptor–Induced Cell Death , 1996, Cell.

[15]  Matthias Mann,et al.  FLICE, A Novel FADD-Homologous ICE/CED-3–like Protease, Is Recruited to the CD95 (Fas/APO-1) Death-Inducing Signaling Complex , 1996, Cell.

[16]  S. Nagata,et al.  Sequential activation of ICE-like and CPP32-like proteases during Fas-mediated apoptosis , 1996, Nature.

[17]  A. Sauter,et al.  Expression of tumor necrosis factor alpha after focal cerebral ischaemia in the rat , 1996, Neuroscience.

[18]  P. Young,et al.  Experimental brain injury induces differential expression of tumor necrosis factor-alpha mRNA in the CNS. , 1996, Brain research. Molecular brain research.

[19]  C. Faucheu,et al.  Identification of a cysteine protease closely related to interleukin-1 beta-converting enzyme. , 1996, European journal of biochemistry.

[20]  P. Caron,et al.  Identification and Characterization of CPP32/Mch2 Homolog 1, a Novel Cysteine Protease Similar to CPP32 (*) , 1996, The Journal of Biological Chemistry.

[21]  A. Chinnaiyan,et al.  ICE-LAP3, a Novel Mammalian Homologue of the Caenorhabditis elegans Cell Death Protein Ced-3 Is Activated during Fas- and Tumor Necrosis Factor-induced Apoptosis (*) , 1996, The Journal of Biological Chemistry.

[22]  A. Faden.  Pharmacological treatment of central nervous system trauma. , 1996, Pharmacology & toxicology.

[23]  T. Tsuruo,et al.  Identification of actin as a substrate of ICE and an ICE-like protease and involvement of an ICE-like protease but not ICE in VP-16-induced U937 apoptosis. , 1995, Biochemical and biophysical research communications.

[24]  E. Alnemri,et al.  Mch3, a novel human apoptotic cysteine protease highly related to CPP32. , 1995, Cancer research.

[25]  D. Bredesen.  Neural apoptosis , 1995, Annals of neurology.

[26]  A. Rosen,et al.  DNA-dependent protein kinase is one of a subset of autoantigens specifically cleaved early during apoptosis , 1995, The Journal of experimental medicine.

[27]  D. Green,et al.  Cell‐free reconstitution of Fas‐, UV radiation‐ and ceramide‐induced apoptosis. , 1995, The EMBO journal.

[28]  S. Nagata,et al.  Apoptosis by a cytosolic extract from Fas‐activated cells. , 1995, The EMBO journal.

[29]  A. Yakovlev,et al.  Molecular Strategies in CNS Injury , 1995 .

[30]  Seamus J. Martin,et al.  Protease activation during apoptosis: Death by a thousand cuts? , 1995, Cell.

[31]  P. Borst,et al.  Antigenic variation in malaria , 1995, Cell.

[32]  Patrick R. Griffin,et al.  Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis , 1995, Nature.

[33]  E. Alnemri,et al.  Mch2, a new member of the apoptotic Ced-3/Ice cysteine protease gene family. , 1995, Cancer research.

[34]  S. Molineaux,et al.  Molecular Cloning and Pro-apoptotic Activity of ICErelII and ICErelIII, Members of the ICE/CED-3 Family of Cysteine Proteases (*) , 1995, The Journal of Biological Chemistry.

[35]  J. Mankovich,et al.  Identification and Characterization of ICH-2, a Novel Member of the Interleukin-1β-converting Enzyme Family of Cysteine Proteases (*) , 1995, The Journal of Biological Chemistry.

[36]  Muneesh Tewari,et al.  Yama/CPP32β, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase , 1995, Cell.

[37]  W. Fiers,et al.  Requirement of an ICE/CED-3 protease for Fas/APO-1-mediated apoptosis , 1995, Nature.

[38]  M. Su,et al.  A novel human protease similar to the interleukin‐1 beta converting enzyme induces apoptosis in transfected cells. , 1995, The EMBO journal.

[39]  M. Chopp,et al.  Temporal Profile of in situ DNA Fragmentation after Transient Middle Cerebral Artery Occlusion in the Rat , 1995, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[40]  U. Tuor,et al.  DNA fragmentation indicative of apoptosis following unilateral cerebral hypoxia-ischemia in the neonatal rat , 1995, Brain Research.

[41]  M. Su,et al.  Altered cytokine export and apoptosis in mice deficient in interleukin-1 beta converting enzyme. , 1995, Science.

[42]  M. Aftabuddin,et al.  Detection of DNA damage induced by apoptosis in the rat brain following incomplete ischemia , 1995, Neuroscience Letters.

[43]  P. Gluckman,et al.  Mechanisms of delayed cell death following hypoxic-ischemic injury in the immature rat: evidence for apoptosis during selective neuronal loss. , 1995, Brain research. Molecular brain research.

[44]  A. Faden,et al.  Neuroprotective effects of 619C89, a use-dependent sodium channel blocker, in rat traumatic brain injury , 1995, Brain Research.

[45]  A. Hara,et al.  Temporal profile of nuclear DNA fragmentation in situ in gerbil hippocampus following transient forebrain ischemia , 1995, Brain Research.

[46]  Z. Werb,et al.  Suppression of ICE and apoptosis in mammary epithelial cells by extracellular matrix , 1995, Science.

[47]  D. Giegel,et al.  Cloning, tissue expression and regulation of rat interleukin 1 beta converting enzyme. , 1995, Cytokine.

[48]  K O'Rourke,et al.  Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. , 1995, Cell.

[49]  M. Chopp,et al.  In situ detection of DNA fragmentation after focal cerebral ischemia in mice. , 1995, Brain research. Molecular brain research.

[50]  E. A. Miranda,et al.  Poly(ADP-ribose) polymerase: structure-function relationship. , 1995, Biochimie.

[51]  J. Trojanowski,et al.  Evidence of apoptotic cell death after experimental traumatic brain injury in the rat. , 1995, The American journal of pathology.

[52]  E. Alnemri,et al.  CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme. , 1994, The Journal of biological chemistry.

[53]  A. Yakovlev,et al.  Sequential expression of c-fos protooncogene, TNF-alpha, and dynorphin genes in spinal cord following experimental traumatic injury. , 1994, Molecular and chemical neuropathology.

[54]  L. Wang,et al.  Ich-1, an Ice/ced-3-related gene, encodes both positive and negative regulators of programmed cell death , 1994, Cell.

[55]  N. Copeland,et al.  Induction of apoptosis by the mouse Nedd2 gene, which encodes a protein similar to the product of the Caenorhabditis elegans cell death gene ced-3 and the mammalian IL-1 beta-converting enzyme. , 1994, Genes & development.

[56]  T. Shiraishi,et al.  Visualization of DNA double strand breaks in the gerbil hippocampal CA1 following transient ischemia , 1994, Neuroscience Letters.

[57]  E. Shohami,et al.  Closed Head Injury Triggers Early Production of TNFα and IL-6 by Brain Tissue , 1994 .

[58]  C. Anderson,et al.  DNA-dependent protein kinase is activated by nicks and larger single-stranded gaps. , 1994, The Journal of biological chemistry.

[59]  R. Lin,et al.  Internucleosomal DNA fragmentation in gerbil hippocampus following forebrain ischemia , 1994, Neuroscience Letters.

[60]  D. Green,et al.  Role of DNA fragmentation in T cell activation-induced apoptosis in vitro and in vivo. , 1994, Journal of immunology.

[61]  M. Fishman,et al.  Prevention of vertebrate neuronal death by the crmA gene. , 1994, Science.

[62]  E. Preston,et al.  Global ischemia can cause DNA fragmentation indicative of apoptosis in rat brain , 1993, Neuroscience Letters.

[63]  M. Linnik,et al.  Evidence Supporting a Role for Programmed Cell Death in Focal Cerebral Ischemia in Rats , 1993, Stroke.

[64]  Junying Yuan,et al.  Induction of apoptosis in fibroblasts by IL-1β-converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3 , 1993, Cell.

[65]  Shai Shaham,et al.  The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1β-converting enzyme , 1993, Cell.

[66]  Y. Lazebnik,et al.  Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis , 1993, The Journal of cell biology.

[67]  J. Cidlowski,et al.  Apoptosis: the biochemistry and molecular biology of programmed cell death. , 1993, Endocrine reviews.

[68]  A. Faden.  Comparison of single and combination drug treatment strategies in experimental brain trauma. , 1993, Journal of neurotrauma.

[69]  K. O. Elliston,et al.  A novel heterodimeric cysteine protease is required for interleukin-1βprocessing in monocytes , 1992, Nature.

[70]  Masahiko S. Satoh,et al.  Role of poly(ADP-ribose) formation in DNA repair , 1992, Nature.

[71]  R. J. Clem,et al.  Prevention of apoptosis by a baculovirus gene during infection of insect cells. , 1991, Science.

[72]  D. Newmeyer,et al.  Egg extracts for nuclear import and nuclear assembly reactions. , 1991, Methods in cell biology.

[73]  W. Bursch,et al.  The biochemistry of cell death by apoptosis. , 1990, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[74]  L. Noble,et al.  Traumatic brain injury in the rat: Characterization of a lateral fluid-percussion model , 1989, Neuroscience.

[75]  R. Vink,et al.  The role of excitatory amino acids and NMDA receptors in traumatic brain injury. , 1989, Science.

[76]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[77]  A. Wyllie,et al.  Cell death: the significance of apoptosis. , 1980, International review of cytology.

[78]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.

[79]  Trump Bf,et al.  Studies of cellular injury in isolated flounder tubules. I. Correlation between morphology and function of control tubules and observations of autophagocytosis and mechanical cell damage. , 1967 .

[80]  B. Trump,et al.  Studies of cellular injury in isolated flounder tubules. I. Correlation between morphology and function of control tubules and observations of autophagocytosis and mechanical cell damage. , 1967, Laboratory investigation; a journal of technical methods and pathology.

[81]  L. Iversen,et al.  REGIONAL STUDIES OF CATECHOLAMINES IN THE RAT BRAIN‐I , 1966, Journal of neurochemistry.