A wound-induced [Ca2+], increase and its transcriptional activation of immediate early genes is important in the regulation of motility

Upon mechanical wounding of a confluent quiescent monolayer, cells move into the denuded zone. However, it is not well known what signaling cascade connects release from contact inhibition to cell movement at the wound edge, Mechanical wounding induced an increase in the concentration of intracellular free Ca2+ ([Ca2+](i)) in endothelial cells at the wound edge, The [Ca2+](i) signal was required for the transcriptional activation of two immediate early genes (IEGs), c-fos and c-jun, since blocking Ca2+ influx with Gd3+ or EGTA reduced IEG transcription, while augmenting Ca2+ influx increased IEG transcription. The transcriptional activation of the IEGs depended on protein kinase C and calmodulin-dependent protein kinase since treatment with the inhibitors C alphostin C and KN-62 significantly reduced IEG expression. Briefly blocking Ca2+ influx also produced a long-term reduction of cell motility, while augmenting Ca2+ influx increased cell motility. To evaluate whether expression of IEGs might control cell movement, we microinjected sense or antisense cDNA to c-fos into cells after wounding. Antisense c-fos cDNA inhibited motility, while sense cDNA increased motility rates. These results suggested that the [Ca2+](i) rise, induced by wounding, regulated the initiation of subsequent motility through the transcriptional activation of IEGs during wounding, (C) 1999 Academic Press.