Thrombomodulin as a new marker of lesion-induced astrogliosis: involvement of thrombin through the G-protein-coupled protease-activated receptor-1.
Because injury of the CNS causes an astrogliosis, characterized by cell swelling and proliferation, similar to the effects of the serine protease thrombin on astrocytes, we hypothesized that a high level of thrombin at the site of injury might initially induce an astrocyte reaction and later increase the expression of its specific inhibitor, thrombomodulin. Thrombomodulin could then stabilize the astroglial scar through its adhesive properties. Here, we studied the in vivo injury response of astrocytes in the anterior medullary velum of adult rat by immunostaining and in situ hybridization of thrombomodulin. Thrombomodulin was poorly expressed on astrocytes in normal tissue, increased up to 2 d after injury, and was still highly expressed at 6 d. To check that thrombin had a direct effect on thrombomodulin expression by astrocytes, we used brain cortical astrocyte primary cultures treated with either thrombin or the agonist peptide thrombin receptor-activating peptide-6, known to activate directly the thrombin G-protein-coupled receptor (GPCR) protease-activated receptor-1 (PAR-1). Modification of thrombomodulin expression was studied by Western blotting and quantitative reverse transcription-PCR. There was a dose-dependent increase in thrombomodulin after 48 hr of treatment, with gene expression peaking at 24 hr but falling to control levels by 48 hr. Together, these results show the following: (1) injury increases astrocyte thrombomodulin expression; (2) thrombin might mediate thrombomodulin expression via the specific receptor PAR-1; and (3) serine proteases, their inhibitors, and the new family of GPCR, PARs, are active on astrogliosis.
Pindon
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Institut National de la Santé et de la Recherche Médicale Unité 523 (formerly 153), Institut de Myologie, Hôpital de la Salpêtrière, F-75013 Paris, France. inserm523@myologie.chups.jussieu.fr
Berry
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M
Hantaï
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D
D000818:Animals; D001253:Astrocytes / Q000502:physiology*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005911:Gliosis / Q000473:pathology / Q000503:physiopathology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin / Q000502:physiology*; D013917:Thrombin / Q000502:physiology*; D018180:Thrombomodulin / Q000032:analysis* / Q000096:biosynthesis; D015854:Up-Regulation
D012333:RNA, Messenger; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin; D018180:Thrombomodulin; D013917:Thrombin; D019204:GTP-Binding Proteins