Thrombin-induced reversal of astrocyte stellation is mediated by activation of protein kinase C beta-1.
Exogenous or endogenous injuries of the central nervous system trigger astrogliosis characterized by proliferation of astrocytes and changes in their morphology from stellate to flat polygonal. Astrocytes in culture are very sensitive to thrombin, a serine protease, which through its proteolytically activated receptor (PAR-1) induces proliferation and morphological changes comparable to astrogliosis. Evaluation of the thrombin signal-transduction pathway in the reversal of astrocyte stellation might help to understand astrogliosis. For this purpose, primary cultured murine cortical astrocytes were treated with H7, a protein-kinase inhibitor, and thrombin, which resulted in an inhibition of stellation reversal. Treatments with phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, mimicked the action of thrombin. Subsequently, direct assay of astrocyte PKC activity after thrombin or PMA treatment demonstrated involvement of PKC in thrombin signaling associated with shape change. Western blotting showed that PKC isoform beta-1 was involved in this pathway, while PKC alpha was only weakly activated and PKC beta-2 was not activated by thrombin. PKC beta-1 translocation was also elicited by a thrombin-receptor active peptide (SFLLRN), demonstrating the involvement of PAR-1 in this process. PKC delta and epsilon were located constitutively in the membrane fraction in stellate astrocytes. Isoforms gamma, eta, theta, and zeta were absent from astrocytes. These results suggest that astrogliosis in vitro might be regulated by modulating the activity of thrombin, PAR-1, or specific PKC isoforms.
Pindon
A
A
Institut National de la Santé et de la Recherche Médicale Unité 153, Hôpital de la Salpêtrière, Paris, France. apindon@myologie.infobiogen.fr
Festoff
B W
BW
Hantaï
D
D
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000502:physiology; D015153:Blotting, Western; D002478:Cells, Cultured; D004789:Enzyme Activation; D007527:Isoenzymes / Q000502:physiology*; D051379:Mice; D010446:Peptide Fragments / Q000494:pharmacology; D011493:Protein Kinase C / Q000502:physiology*; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology*
D007527:Isoenzymes; D010446:Peptide Fragments; C082835:thrombin receptor peptide (42-47); D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D013917:Thrombin; D013755:Tetradecanoylphorbol Acetate