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Volume 17, Number 16, Issue of August 15, 1997 pp. 6203-6212
Copyright ©1997 Society for NeuroscienceGrowth Factor Activity of Endothelin-1 in Primary Astrocytes Mediated by Adhesion-Dependent and -Independent Pathways
Received Jan. 7, 1997; revised May 19, 1997; accepted May 29, 1997.
Sylvie Cazaubon1, Nathalie Chaverot1, Ignacio A. Romero1, Jean-Antoine Girault2, Peter Adamson3, A. Donny Strosberg1, and Pierre-Olivier Couraud1 1 Centre National de la Recherche Scientifique UPR 0415, Institut Cochin de Génétique Moléculaire, 75014 Paris, France, 2 Institut National de la Santé et de la Recherche Médicale U114, Collège de France, 75005 Paris, France, and 3 Institute of Ophthalmology, University College London, London EC1V 9EL, United Kingdom
Endothelin-1 (ET-1) has been shown to induce DNA synthesis in primary astrocytes by stimulating the extracellular signal-regulated kinase (ERK) pathway. To clarify the mechanisms responsible for the anchorage-dependent growth of astrocytes, the relationships between cell adhesion and ERK activation were investigated. Here it is reported that ET-1 promotes the formation of stress fibers and focal adhesions and the tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin, as well as Src activation and association of phosphorylated FAK with Grb2. Pretreatment of astrocytes with cytochalasin D or C3-transferase, which inhibits actin polymerization or Rho activity, respectively, prevented the activation/phosphorylation of Src, FAK, and paxillin after ET-1 stimulation; by contrast, the ERK pathway was not significantly affected. This differential activation of FAK/Src and ERK pathways was also observed with astrocytes 10 and 60 min after replating on poly-L-ornithine-precoated dishes. Collectively, these findings indicate that activation of FAK and Src is dependent on actin cytoskeleton integrity, Rho activation, and adhesion to extracellular matrix, whereas ERK activation is independent of these intracellular events and seems to correlate with activation of the newly identified protein tyrosine kinase PYK2. Induction of DNA synthesis by ET-1, however, was reduced dramatically in astrocytes pretreated with either cytochalasin D or C3-transferase. This study provides a demonstration of Rho- and adhesion-dependent activation of FAK/Src, which collaborates with adhesion-independent activation of PYK2/ERK for DNA synthesis in ET-1-stimulated astrocytes.
Key words: endothelin-1; growth factor; primary astrocyte; cell adhesion; stress fibers; focal adhesion kinase; extracellular signal-regulated kinase
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