alpha(1)-Adrenergic receptor stimulation of cell motility requires phospholipase D-mediated extracellular signal-regulated kinase activation.
Phospholipase D is suspected to play a role in tumorigenesis, and the inhibition of phospholipase D has been associated with changes in several cellular events including invasion and migration. We report here that the specific alpha(1)-adrenergic receptor agonist, phenylepherine, signals to a growth factor pathway in a manner that requires phospholipase D activity in CCL39 fibroblasts. Phenylepherine increased extracellular signal-regulated kinase phosphorylation eightfold and promoted stress fiber formation threefold. Stress fiber formation was blocked when extracellular signal-regulated kinase activation was inhibited. Stimulation of CCL39 fibroblasts by phenylepherine increased the rate of wound healing fourfold in a wounding assay, while treatment with the MEK inhibitor, PD98059 reduced the closure of phenylepherine-induced wound healing to control levels. Addition of 1-butanol but not 2-butanol inhibited extracellular signal-regulated kinase activation by phenylepherine, presumably by blocking the formation of phosphatidic acid. Exogenously added cell permeable phosphatidic acid increased extracellular signal-regulated kinase activation in a time- and dose-dependent manner as well as stimulated the formation of stress fibers. 1-butanol also significantly inhibited the ability of phenylepherine to stimulate stress fiber formation and wound healing. Taken together, these results indicate a novel role for phospholipase D in the activation of the extracellular signal-regulated kinase growth factor pathway to stimulate early cellular events induced by phenylepherine.
Sang
Rachel L
RL
Department of Bioscience, Minnesota State University Moorhead, Moorhead, MN 56563, USA.
Johnson
Jessica F
JF
Taves
Jennifer
J
Nguyen
Callie
C
Wallert
Mark A
MA
Provost
Joseph J
JJ
D058646:Adrenergic alpha-1 Receptor Agonists; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects; D002465:Cell Movement; D006224:Cricetinae; D003412:Cricetulus; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D020404:Glycerophospholipids / Q000096:biosynthesis; D015394:Molecular Structure; D010656:Phenylephrine / Q000494:pharmacology; D010739:Phospholipase D / Q000378:metabolism*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism*; D014945:Wound Healing
D058646:Adrenergic alpha-1 Receptor Agonists; D020404:Glycerophospholipids; D047428:Protein Kinase Inhibitors; D018340:Receptors, Adrenergic, alpha-1; D010656:Phenylephrine; D048049:Extracellular Signal-Regulated MAP Kinases; D010739:Phospholipase D