title: Reduction of glutamate-induced excitotoxicity in murine primary neurons involving calpain inhibition.
abstract: Excessive glutamate secretion leads to excitotoxicity, which has been shown to underlie neurodegenerative disorders. Excitotoxicity is in part exerted by overactivation of calpains, which promote neuronal cell death via induction of limited proteolysis of the cellular proteins p35, regulatory subunit of cyclin-dependent kinase 5, and αII-spectrin. We used primary murine neuronal cells in a model of glutamate toxicity. The protease inhibitor α1-antitrypsin was able to prevent glutamate toxicity as determined by MTT assay and immunofluorescence. Calpain and caspase 3 activity were reduced following α1-antitrypsin treatment, as assessed by calpain and caspase 3 activity assays. In addition we could observe a modulation of cleavage of the calpain/caspase substrates αII-spectrin and p35 in Western blots. In summary, α1-antitrypsin shows inhibitory effects on excitotoxicity of primary neurons involving the inhibition of calpain activity. The advantage of using α1-antitrypsin is that the substance is already in clinical use for the treatment of patients with hereditary α1-antitrypsin deficiency. Further experiments are required in animal models of neurodegenerative disorders to assess the suitability of this substance in patients suffering from Alzheimer's disease or Parkinson's disease.
authors.affiliation: Department of Neurology, University of Marburg, 35043 Marburg, Germany.
authors.forename: Maike
authors.initials: M
authors.lastname: Gold
authors.affiliation: Department of Medicine, Pulmonary and Critical Care Medicine, University of Marburg, 35043 Marburg, Germany.
authors.forename: Andreas-Rembert
authors.initials: AR
authors.lastname: Koczulla
authors.affiliation: Department of Neurology, University of Marburg, 35043 Marburg, Germany.
authors.forename: David
authors.initials: D
authors.lastname: Mengel
authors.affiliation: Department of Medicine, Pulmonary and Critical Care Medicine, University of Marburg, 35043 Marburg, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Koepke
authors.affiliation: Department of Neurology, University of Marburg, 35043 Marburg, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Dodel
authors.affiliation: Department of Neurology, RWTH Aachen, 52074 Aachen, Germany.
authors.forename: Guergana
authors.initials: G
authors.lastname: Dontcheva
authors.affiliation: Department of Neurology, RWTH Aachen, 52074 Aachen, Germany.
authors.forename: Pardes
authors.initials: P
authors.lastname: Habib
authors.affiliation: Department of Neurology, RWTH Aachen, 52074 Aachen, Germany. Electronic address: jbach@ukaachen.de.
authors.forename: Jan-Philipp
authors.initials: JP
authors.lastname: Bach
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002154:Calpain / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018846:Excitatory Amino Acids / Q000633:toxicity*; D018698:Glutamic Acid / Q000633:toxicity*; D051379:Mice; D009474:Neurons / Q000187:drug effects*; D010770:Phosphotransferases / Q000378:metabolism; D013997:Time Factors; D000515:alpha 1-Antitrypsin / Q000494:pharmacology
chemical_list: C492236:Cdk5r1 protein, mouse; D003598:Cytoskeletal Proteins; D004791:Enzyme Inhibitors; C505599:Erc2 protein, mouse; D018846:Excitatory Amino Acids; D000515:alpha 1-Antitrypsin; D018698:Glutamic Acid; D010770:Phosphotransferases; D002154:Calpain; D053148:Caspase 3
keywords: Alzheimer's disease; Calpain; Cell death; Excitotoxicity; Neuroprotection; α1-Antitrypsin