Regulation of motility in bovine brain endothelial cells.
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Rosen EM, Jaken S, Carley W, Luckett PM, Setter E, Bhargava M, Goldberg ID
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Pediatric Medicine
Emergency Medicine
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Emergency Medicine
Pediatric Medicine
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volume
146
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0021-9541
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["Rosen EM, Jaken S, Carley W, Luckett PM, Setter E, Bhargava M, Goldberg ID","Animals, Cattle, Cell Division drug effects, Cell Movement drug effects, Cell Movement physiology, Cytoskeleton drug effects, Down-Regulation, Endothelium, Vascular cytology, Growth Substances physiology, Hepatocyte Growth Factor, In Vitro Techniques, Nucleotides, Cyclic physiology, Phorbol Esters pharmacology, Protein Kinase Inhibitors, Signal Transduction physiology, Time Factors, Brain blood supply, Endothelium, Vascular physiology, Proteins physiology","English","1991-02-01","Journal of cellular physiology","146"]
Description
Scatter factor (SF) is a fibroblast-derived cytokine which stimulates motility of epithelial and vascular endothelial cells. We used a quantitative assay based on migration of cells from microcarrier beads to flat surfaces to study the regulation of motility in bovine brain endothelial cells (BBEC). Peptide growth factors (EGF, ECGF, basic FGF) did not stimulate migration. Tumor promoting phorbol esters (PMA, PDD) markedly stimulated migration, while inactive phorbol esters (4a-PDD, phorbol-13,20-diacetate) did not affect migration. Both SF- and PMA-stimulated migration were inhibited by 1) TGF-beta; 2) protein kinase inhibitors (e.g., staurosporine, K-252a); 3) activators of the adenylate cyclase signaling pathway (e.g., dibutyryl cyclic AMP, theophylline); 4) cycloheximide; and 5) anti-cytoskeleton agents (e.g., cytochalasin B, colcemid). However, PMA and SF pathways were distinguishable: 1) PMA induced additional migration at saturating SF concentrations; 2) the onset of migration-stimulation was immediate for PMA and delayed for SF; and 3) down-modulation of protein kinase C (PKC) ablated PMA but not SF responsiveness. Assessment of PKC by (3H)-phorbol ester (PDBu) binding and by immunoblot showed 1) scatter factor does not cause significant redistribution or down-modulation of PDBu binding or alpha-PKC; and 2) PDBu mediates redistribution and down-modulation of both binding and alpha-PKC. These findings suggest two pathways for BBEC motility: a PKC-dependent pathway and an SF-stimulated/PKC-independent pathway.
