Caspase-Mediated Suppression of Glutamate (AMPA) Receptor Channel Activity in Hippocampal Neurons in Response to DNA Damage Promotes Apoptosis and Prevents Necrosis: Implications for Neurological Side Effects of Cancer Therapy and Neurodegenerative Disorders

DNA damage in neurons is implicated in the pathogenesis of several neurodegenerative disorders and may also contribute to the often severe neurological complications in cancer patients treated with chemotherapeutic agents. DNA damage can trigger apoptosis, a form of controlled cell death that involves activation of cysteine proteases called caspases. The excitatory neurotransmitter glutamate plays central roles in the activation of neurons and in processes such as learning and memory, but overactivation of ionotropic glutamate receptors can induce either apoptosis or necrosis. Glutamate receptors of the AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate) type mediate such physiological and pathological processes in most neurons. We now report that DNA damage can alter glutamate receptor channel activity by a mechanism involving activation of caspases. Whole-cell patch clamp analyses revealed a marked decrease in AMPA-induced currents after exposure of neurons to camptothecin, a topoisomerase inhibitor that induces DNA damage; N-methyl-d-aspartate (NMDA)-induced currents were unaffected by camptothecin. The decrease in AMPA-induced current was accompanied by a decreased calcium response to AMPA. Pharmacological inhibition of caspases abolished the effects of camptothecin on AMPA-induced current and calcium responses, and promoted excitotoxic necrosis. Combined treatment with glutamate receptor antagonists and a caspase inhibitor prevented camptothecin-induced neuronal death. Caspase-mediated suppression of AMPA currents may allow neurons with damaged DNA to withdraw their participation in excitatory circuits and undergo apoptosis, thereby avoiding widespread necrosis. These findings have important implications for treatment of patients with cancer and neurodegenerative disorders.

[1]  C. Culmsee,et al.  A synthetic inhibitor of p53 protects neurons against death induced by ischemic and excitotoxic insults, and amyloid β‐peptide , 2001 .

[2]  J. Pfeilschifter,et al.  New insights into the mechanism for clearance of apoptotic cells. , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  C. Culmsee,et al.  Homocysteine Elicits a DNA Damage Response in Neurons That Promotes Apoptosis and Hypersensitivity to Excitotoxicity , 2000, The Journal of Neuroscience.

[4]  P. Dash,et al.  Caspase activity plays an essential role in long‐term memory , 2000, Neuroreport.

[5]  M. Mattson,et al.  Caspase-Mediated Degradation of AMPA Receptor Subunits: A Mechanism for Preventing Excitotoxic Necrosis and Ensuring Apoptosis , 2000, The Journal of Neuroscience.

[6]  David S. Park,et al.  Involvement of Retinoblastoma Family Members and E2F/DP Complexes in the Death of Neurons Evoked by DNA Damage , 2000, The Journal of Neuroscience.

[7]  J. Schulz,et al.  Neuroprotection by the Inhibition of Apoptosis , 2000, Brain pathology.

[8]  S. Snyder,et al.  Poly(ADP-ribosyl)ation basally activated by DNA strand breaks reflects glutamate-nitric oxide neurotransmission. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. Murguía,et al.  Sensing and responding to DNA damage. , 2000, Current opinion in genetics & development.

[10]  M. Maki,et al.  Caspases cleave the amino-terminal calpain inhibitory unit of calpastatin during apoptosis in human Jurkat T cells. , 2000, Journal of biochemistry.

[11]  J. Vonsattel,et al.  DNA strand breaks in Alzheimer's disease , 1999, Brain Research.

[12]  T. Sugawara,et al.  Early decrease of XRCC1, a DNA base excision repair protein, may contribute to DNA fragmentation after transient focal cerebral ischemia in mice. , 1999, Stroke.

[13]  M. Mattson,et al.  ALS-Linked Cu/Zn–SOD Mutation Increases Vulnerability of Motor Neurons to Excitotoxicity by a Mechanism Involving Increased Oxidative Stress and Perturbed Calcium Homeostasis , 1999, Experimental Neurology.

[14]  M. Mattson,et al.  Caspase and calpain substrates: Roles in synaptic plasticity and cell death , 1999, Journal of neuroscience research.

[15]  M V Chernov,et al.  A chemical inhibitor of p53 that protects mice from the side effects of cancer therapy. , 1999, Science.

[16]  David S. Park,et al.  Caspase-Dependent and -Independent Death of Camptothecin-Treated Embryonic Cortical Neurons , 1999, The Journal of Neuroscience.

[17]  M. Mattson,et al.  Evidence for caspase‐mediated cleavage of AMPA receptor subunits in neuronal apoptosis and Alzheimer's disease , 1999 .

[18]  P. Brundin,et al.  Excitotoxicity Plays a Role in the Death of Tyrosine Hydroxylase- Immunopositive Nigral Neurons Cultured in Serum-Free Medium , 1999, Experimental Neurology.

[19]  S. Snyder,et al.  Poly (ADP-ribose) polymerase, nitric oxide and cell death. , 1999, Trends in pharmacological sciences.

[20]  T. Whiteside,et al.  Caspase-mediated Degradation of T-Cell Receptor z-Chain1 , 1999 .

[21]  G. Ross.  Induction of cell death by radiotherapy. , 1999, Endocrine-related cancer.

[22]  Y. Pommier,et al.  Role of oxygen radicals generated by NADPH oxidase in apoptosis induced in human leukemia cells. , 1998, The Journal of clinical investigation.

[23]  Shahrooz Rabizadeh,et al.  The DCC gene product induces apoptosis by a mechanism requiring receptor proteolysis , 1998, Nature.

[24]  J. Dichgans,et al.  Extended therapeutic window for caspase inhibition and synergy with MK-801 in the treatment of cerebral histotoxic hypoxia , 1998, Cell Death and Differentiation.

[25]  C. Cotman,et al.  GADD45Is Induced in Alzheimer's Disease, and Protects against Apoptosisin Vitro , 1998, Neurobiology of Disease.

[26]  S. Kaufmann.  Cell death induced by topoisomerase-targeted drugs: more questions than answers. , 1998, Biochimica et biophysica acta.

[27]  S. Lipton,et al.  Role of Caspases in N‐Methyl‐d‐Aspartate‐Induced Apoptosis in Cerebrocortical Neurons , 1998, Journal of neurochemistry.

[28]  M. Mattson,et al.  Evidence for Synaptic Apoptosis , 1998, Experimental Neurology.

[29]  M. Mattson,et al.  The Transcription Factor NF‐κB Mediates Increases in Calcium Currents and Decreases in NMDA‐ and AMPA/Kainate‐Induced Currents Induced by Tumor Necrosis Factor‐α in Hippocampal Neurons , 1998, Journal of neurochemistry.

[30]  S. Love,et al.  Apoptosis and expression of DNA repair proteins in ischaemic brain injury in man , 1998, Neuroreport.

[31]  E. Schmitt,et al.  Bcl-xL modulates apoptosis induced by anticancer drugs and delays DEVDase and DNA fragmentation-promoting activities. , 1998, Experimental cell research.

[32]  E. Michaelis.  Molecular biology of glutamate receptors in the central nervous system and their role in excitotoxicity, oxidative stress and aging , 1998, Progress in Neurobiology.

[33]  David S. Park,et al.  Multiple Pathways of Neuronal Death Induced by DNA-Damaging Agents, NGF Deprivation, and Oxidative Stress , 1998, The Journal of Neuroscience.

[34]  A. Khar,et al.  Caspase-mediated apoptosis in AK-5 tumor cells: a cell-free study using peptide inhibitors and antisense strategy. , 1997, Experimental cell research.

[35]  M. Mattson,et al.  The Actin-Severing Protein Gelsolin Modulates Calcium Channel and NMDA Receptor Activities and Vulnerability to Excitotoxicity in Hippocampal Neurons , 1997, The Journal of Neuroscience.

[36]  M. Moskowitz,et al.  Ischemic Brain Injury is Mediated by the Activation of Poly(ADP-Ribose)Polymerase , 1997, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[37]  P. C. Staton,et al.  The Dietary Excitotoxins β‐N‐Methylamino‐l‐Alanine and β‐N‐Oxalylamino‐l‐Alanine Induce Necrotic‐ and Apoptotic‐Like Death of Rat Cerebellar Granule Cells , 1997 .

[38]  C D Marsden,et al.  Oxidative DNA Damage in the Parkinsonian Brain: An Apparent Selective Increase in 8‐Hydroxyguanine Levels in Substantia Nigra , 1997, Journal of neurochemistry.

[39]  M. Rothenberg.  Topoisomerase I inhibitors: review and update. , 1997, Annals of oncology : official journal of the European Society for Medical Oncology.

[40]  P. Beart,et al.  Apoptosis Induced via AMPA‐Selective Glutamate Receptors in Cultured Murine Cortical Neurons , 1997, Journal of neurochemistry.

[41]  M. Mattson,et al.  4-Hydroxynonenal, an aldehydic product of membrane lipid peroxidation, impairs glutamate transport and mitochondrial function in synaptosomes , 1997, Neuroscience.

[42]  R. Miller,et al.  Ca2+ and Reactive Oxygen Species in Staurosporine‐Induced Neuronal Apoptosis , 1997, Journal of neurochemistry.

[43]  K. Wilson,et al.  Substrate and Inhibitor Specificity of Interleukin-1β-converting Enzyme and Related Caspases* , 1997, The Journal of Biological Chemistry.

[44]  R J Mark,et al.  Amyloid β-Peptide Impairs Glucose Transport in Hippocampal and Cortical Neurons: Involvement of Membrane Lipid Peroxidation , 1997, The Journal of Neuroscience.

[45]  C. Cotman,et al.  Bax Protein Expression Is Increased in Alzheimer's Brain: Correlations with DNA Damage, Bcl‐2 Expression, and Brain Pathology , 1997, Journal of neuropathology and experimental neurology.

[46]  H. M. Geller,et al.  Induction of neuronal apoptosis by camptothecin, an inhibitor of DNA topoisomerase-I: evidence for cell cycle-independent toxicity , 1996, The Journal of cell biology.

[47]  S. Lipton,et al.  Glutamate-induced neuronal death: A succession of necrosis or apoptosis depending on mitochondrial function , 1995, Neuron.

[48]  T. Cotter,et al.  Inhibition of apoptosis by antioxidants in the human HL-60 leukemia cell line. , 1995, Biochemical pharmacology.

[49]  S. Snyder,et al.  Poly(ADP‐Ribose) Synthetase Activation: An Early Indicator of Neurotoxic DNA Damage , 1995, Journal of neurochemistry.

[50]  M. Mattson,et al.  Amyloid beta-peptide impairs ion-motive ATPase activities: evidence for a role in loss of neuronal Ca2+ homeostasis and cell death , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[51]  A. Doble.  Excitatory amino acid receptors and neurodegeneration. , 1995, Therapie.

[52]  M. Chopp,et al.  Induction of DNA fragmentation after 10 to 120 minutes of focal cerebral ischemia in rats. , 1995, Stroke.

[53]  T. Tsuruo,et al.  Aspartate-based inhibitor of interleukin-1 beta-converting enzyme prevents antitumor agent-induced apoptosis in human myeloid leukemia U937 cells. , 1995, Biochemical and biophysical research communications.

[54]  Mark P. Mattson,et al.  β-Amyloid precursor protein metabolites and loss of neuronal Ca2+ homeostasis in Alzheimer's disease , 1993, Trends in Neurosciences.

[55]  N. Akaike,et al.  Hyperpolarizing muscarinic responses of freshly dissociated rat hippocampal CA1 neurones. , 1993, The Journal of physiology.

[56]  D. Choi.  Excitotoxic cell death. , 1992, Journal of neurobiology.

[57]  J. Backon.  Dementia in cancer patients undergoing chemotherapy: implication of free radical injury and relevance to Alzheimer disease. , 1991, Medical hypotheses.

[58]  W. Bradley,et al.  DNA damage and chronic neuronal degenerations , 1984, Journal of the Neurological Sciences.

[59]  Miles W. Miller,et al.  Increased vulnerability of hippocampal neurons to excitotoxic necrosis in presenilin-1 mutant knock-in mice , 1999, Nature Medicine.

[60]  S. Arbuck,et al.  DNA topoisomerase I poisons. , 1999, Cancer chemotherapy and biological response modifiers.

[61]  M. Baudry,et al.  Posttranslational regulation of ionotropic glutamate receptors and synaptic plasticity. , 1998, International review of neurobiology.

[62]  G. Kroemer,et al.  The mitochondrial death/life regulator in apoptosis and necrosis. , 1998, Annual review of physiology.

[63]  D. Meyer,et al.  The CrmA- and TPCK-sensitive pathways that trigger oligonucleosome-sized DNA fragmentation in camptothecin-induced apoptosis: relation to caspase activation and high molecular weight DNA fragmentation. , 1997, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[64]  S. Garattini,et al.  DNA topoisomerase II poisons and inhibitors. , 1997, Cancer chemotherapy and biological response modifiers.

[65]  M. Mattson,et al.  Calcium, free radicals, and excitotoxic neuronal death in primary cell culture. , 1995, Methods in cell biology.

[66]  J. Rothstein.  Excitotoxic mechanisms in the pathogenesis of amyotrophic lateral sclerosis. , 1995, Advances in neurology.

[67]  M. Hollmann,et al.  Molecular neurobiology of glutamate receptors. , 1992, Annual review of physiology.