title: Alterations in the Cerebral Microvascular Proteome Expression Profile After Transient Global Cerebral Ischemia in Rat.
abstract: This study aimed at obtaining an in-depth mapping of expressional changes of the cerebral microvasculature after transient global cerebral ischemia (GCI) and the impact on these GCI-induced expressional changes of post-GCI treatment with a mitogen-activated protein kinase kinase (MEK1/2) inhibitor. GCI was induced in male Wistar rats followed by treatment with either vehicle or the MEK1/2 inhibitor U0126 every 12 h post-GCI. Seventy-two hours after GCI or sham surgery, the cerebral microvasculature was isolated and the protein content analysed with state-of-the-art mass spectrometry. The proteomic profile of the isolated cerebral microvasculature 72 h after GCI (compared to sham) indicated that the main expressional changes could be divided into nine categories: (1) cellular respiration, (2) remodelling of the extracellular matrix, (3) decreased contractile phenotype, (4) clathrin-mediated endocytosis, (5) ribosomal activity, (6) expression of chromatin structure-related proteins, (7) altered synaptic activity, (8) altered G-protein signalling and (9) instability of the membrane potential. Treatment with U0126 partly normalized the expression of one or more of the proteins in all nine categories. Flow cytometry confirmed key findings from the proteome such as upregulation of the extracellular proteins lamininβ2 and nidogen2 (p < 0.05) after GCI. These results provide valuable molecular insight into the broad and complex expressional changes in the cerebral microvasculature after GCI and the effect of early MEK1/2 inhibitor treatment on these changes.
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Rigshospitalet, Glostrup, Denmark. stine.spray@regionh.dk.
authors.forename: Stine
authors.identifier: http://orcid.org/0000-0002-9381-7209
authors.initials: S
authors.lastname: Spray
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Rigshospitalet, Glostrup, Denmark.
authors.forename: Sara E
authors.initials: SE
authors.lastname: Johansson
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
authors.forename: Alistair V G
authors.initials: AV
authors.lastname: Edwards
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
authors.forename: Martin R
authors.initials: MR
authors.lastname: Larsen
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Rigshospitalet, Glostrup, Denmark.
authors.forename: Aneta
authors.initials: A
authors.lastname: Radziwon-Balicka
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Rigshospitalet, Glostrup, Denmark.
authors.forename: Gro K
authors.initials: GK
authors.lastname: Povlsen
authors.affiliation: Department of Clinical Experimental Research, Glostrup Research Institute, Rigshospitalet, Glostrup, Denmark.|Department of Clinical Sciences, Division of Experimental Vascular Research, Lund University, Lund, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Edvinsson
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism; D007797:Laminin / Q000235:genetics / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D055806:Microvessels / Q000378:metabolism*; D020543:Proteome / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D007797:Laminin; D008562:Membrane Glycoproteins; D020543:Proteome; C029967:nidogen; C058783:laminin beta2
keywords: Cerebral microvasculature; Extracellular matrix remodelling; Global cerebral ischemia; MEK1/2 inhibitor treatment; Proteomics