Vascular endothelial growth factor stimulates neurite outgrowth from cerebral cortical neurons via Rho kinase signaling

Vascular endothelial growth factor (VEGF), which is prominently involved in angiogenesis, also exerts direct effects on neurons, leading to neurite extension, neuroprotection, and neurogenesis. However, the signal transduction pathways employed by VEGF in neurons are incompletely understood. We investigated the molecular mechanisms through which VEGF stimulates neurogenesis in primary cultures of rat cerebral cortical neurons. VEGF increased neurite outgrowth, measured using a colorimetric assay for cresyl violet staining of neuronal processes, with half-maximal enhancement at 10 ng/mL and maximal, ≈60% enhancement at 30-100 ng/mL. The effect of VEGF was not reproduced by VEGF-B or placental growth factor, but was blocked by SU1498, consistent with a VEGFR2 receptor-mediated process. VEGF-induced neurite outgrowth was also blocked by the ROK inhibitor Y27632 and the Rho inhibitors sulindac and Clostridium botuliunt exoenzyme C3, and was accompanied by Y27632-sensitive phosphorylation of cofilin, a downstream mediator of Rho/ROK signaling. We conclude that VEGF promotes neurite outgrowth from cerebral cortical neurons by interacting with VEGFR2 and activating Rho/ROK signaling pathways.