Inflammation and dephosphorylation of the tight junction protein occludin in an experimental model of multiple sclerosis.
Multiple sclerosis (MS) is a disease of the CNS in which inflammation, demyelination and neurodegeneration contribute to its initiation and progression. A frequently employed model of MS is experimental autoimmune encephalomyelitis (EAE). Here, to gain new insights into the disease process, an analysis of proteins in extracts of lumbar spinal cord from naïve and EAE rats was undertaken. The data mainly confirm that inflammation and blood-brain barrier (BBB) breakdown are the major hallmarks of disease in this model. Given their importance in the BBB, junctional proteins were further investigated. Occludin, a protein localizing to tight junctions in brain endothelial cells, showed strikingly increased migration in EAE when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This increased migration was mimicked by in vitro phosphatase treatment, implying its dephosphorylation in EAE. Occludin dephosphorylation coincided with the onset of inflammation, slightly preceding visible signs of disease, and was just prior to apparent changes in BBB permeability. These findings suggest occludin is a target for signaling processes in EAE, perhaps regulating the response of the BBB to the inflammatory environment as seen in MS.
Morgan
L
L
Eisai London Research Laboratories Ltd., University College London, London, UK.
Shah
B
B
Rivers
L E
LE
Barden
L
L
Groom
A J
AJ
Chung
R
R
Higazi
D
D
Desmond
H
H
Smith
T
T
Staddon
J M
JM
D000818:Animals; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004660:Encephalitis / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000150:complications / Q000378:metabolism* / Q000473:pathology*; D042783:Endothelial Cells / Q000166:cytology; D005260:Female; D047468:Immunoprecipitation / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000378:metabolism*; D062793:Occludin; D010744:Phosphoric Monoester Hydrolases / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D011917:Rats, Inbred Lew; D013116:Spinal Cord / Q000473:pathology; D019108:Tight Junctions / Q000378:metabolism
D008565:Membrane Proteins; D062793:Occludin; C568987:Ocln protein, rat; D010744:Phosphoric Monoester Hydrolases