title: Glutamate receptor antagonists inhibit calpain-mediated cytoskeletal proteolysis in focal cerebral ischemia.
abstract: Excitatory amino acids may promote microtubular proteolysis observed in ischemic neuronal degeneration by calcium-mediated activation of calpain, a neutral protease. We tested this hypothesis in an animal model of focal cerebral ischemia without reperfusion. Spontaneously hypertensive rats were treated with 2, 3-dihydroxy-6-nitro-7-sulfamoyl-benzo-(F)quinoxaline (NBQX), a competitive antagonist of the neuronal receptor for alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), or cis-4-[phosphono-methyl]-2-piperidine carboxylic acid (CGS 19755), a competitive antagonist of the N-methyl-d-aspartate (NMDA) receptor. After treatment, all animals were subjected to permanent occlusion of the middle cerebral artery for 6 or 24 h. Infarct volumes measured in animals pretreated with CGS 19755 after 24 h of ischemia were significantly smaller than those quantified in ischemic controls. Rats pretreated with NBQX showed partial amelioration of cytoskeletal injury with preserved immunolabeling of microtubule-associated protein 2 (MAP 2) at 6 and 24 h and reduced accumulation of calpain-cleaved spectrin byproducts only at 6 h. Prevention of cytoskeletal damage was more effective after pretreatment with CGS 19755, as shown by retention of MAP 2 immunolabeling and significant restriction of calpain activity at both 6 and 24 h. Preserved immunolabeling of tau protein was observed at 6 and 24 h only in animals pretreated with CGS 19755. Western analysis performed on ischemic cortex taken from controls or rats pretreated with either NBQX or CGS 19755 suggested that loss of tau protein immunoreactivity was caused by dephosphorylation, rather than proteolysis. These results demonstrate a crucial link between excitotoxic neurotransmission, microtubular proteolysis, and neuronal degeneration in focal cerebral ischemia.
authors.affiliation: The Stroke Program of the University of Kentucky Chandler Medical Center, USA.
authors.forename: S L
authors.initials: SL
authors.lastname: Minger
authors.forename: J W
authors.initials: JW
authors.lastname: Geddes
authors.forename: M L
authors.initials: ML
authors.lastname: Holtz
authors.forename: S D
authors.initials: SD
authors.lastname: Craddock
authors.forename: S W
authors.initials: SW
authors.lastname: Whiteheart
authors.forename: R G
authors.initials: RG
authors.lastname: Siman
authors.forename: L C
authors.initials: LC
authors.lastname: Pettigrew
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002154:Calpain / Q000037:antagonists & inhibitors*; D002544:Cerebral Infarction / Q000503:physiopathology; D003599:Cytoskeleton / Q000187:drug effects / Q000201:enzymology*; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology*; D007150:Immunohistochemistry; D002546:Ischemic Attack, Transient / Q000201:enzymology*; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D010875:Pipecolic Acids / Q000494:pharmacology; D011810:Quinoxalines / Q000494:pharmacology; D051381:Rats; D011918:Rats, Inbred SHR; D017470:Receptors, Glutamate / Q000187:drug effects*; D013049:Spectrin / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D016875:tau Proteins / Q000378:metabolism
chemical_list: D018691:Excitatory Amino Acid Antagonists; D008869:Microtubule-Associated Proteins; D010875:Pipecolic Acids; D011810:Quinoxalines; D017470:Receptors, Glutamate; D016875:tau Proteins; C062865:2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline; D013049:Spectrin; C053672:selfotel; D002154:Calpain