title: Neuroproteome changes after ischemia/reperfusion injury and tissue plasminogen activator administration in rats: a quantitative iTRAQ proteomics study.
abstract: The thrombolytic, recombinant tissue plasminogen activator (rt-PA) is the only approved therapy for acute ischemic stroke (AIS). When administered after AIS, rt-PA has many adverse pleiotropic actions, which are currently poorly understood. The identification of proteins showing differential expression after rt-PA administration may provide insight into these pleiotropic actions. In this study we used a 2D-LC MS/MS iTRAQ proteomic analysis, western blotting, and pathway analysis to analyze changes in protein expression 24-hours after rt-PA administration in the cortical brain tissue of 36 rats that underwent a sham or transient middle cerebral artery occlusion surgery. After rt-PA administration we reported alterations in the expressions of 18 proteins, many of which were involved in excitatory neurotransmitter function or cytoskeletal structure. The expression changes of GAD2 and EAAT1 were validated with western blot. The interactions between the identified proteins were analyzed with the IPA pathway analysis tool and three proteins: DPYSL2, RTN4, and the NF-kB complex, were found to have characteristics of being key proteins in the network. The differential protein expressions we observed may reflect pleiotropic actions of rt-PA after experimental stroke, and shine light on the mechanisms of rt-PA's adverse effects. This may have important implications in clinical settings where thrombolytic therapy is used to treat AIS.
authors.affiliation: Physiology and Experimental Medicine, The Hospital For Sick Children, Toronto, Ontario, Canada.
authors.forename: Zamir
authors.initials: Z
authors.lastname: Merali
authors.affiliation: Department of Medical Imaging, University Of Toronto, Toronto, Ontario, Canada.
authors.forename: Meah MingYang
authors.initials: MM
authors.lastname: Gao
authors.affiliation: Department of Medical Imaging, University Of Toronto, Toronto, Ontario, Canada.
authors.forename: Tim
authors.initials: T
authors.lastname: Bowes
authors.affiliation: Ontario Cancer Biomarker Network, Toronto, Ontario Canada.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.affiliation: Ontario Cancer Biomarker Network, Toronto, Ontario Canada.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Evans
authors.affiliation: Department of Medical Imaging, University Of Toronto, Toronto, Ontario, Canada; Physiology and Experimental Medicine, The Hospital For Sick Children, Toronto, Ontario, Canada.
authors.forename: Andrea
authors.initials: A
authors.lastname: Kassner
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism; D027341:Excitatory Amino Acid Transporter 1 / Q000235:genetics / Q000378:metabolism; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism; D020244:Infarction, Middle Cerebral Artery / Q000235:genetics / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D009185:Myelin Proteins / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000235:genetics / Q000378:metabolism; D000070798:Nogo Proteins; D020543:Proteome / Q000235:genetics* / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D040641:Quantitative Trait Loci / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015427:Reperfusion Injury / Q000235:genetics* / Q000378:metabolism*; D020521:Stroke / Q000235:genetics / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000235:genetics* / Q000378:metabolism*
chemical_list: D027341:Excitatory Amino Acid Transporter 1; D036341:Intercellular Signaling Peptides and Proteins; D009185:Myelin Proteins; D016328:NF-kappa B; D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; D000070798:Nogo Proteins; D020543:Proteome; D011994:Recombinant Proteins; C000603197:Rtn4 protein, rat; C493392:Slc1a3 protein, rat; C095114:collapsin response mediator protein-2; D010959:Tissue Plasminogen Activator; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2