BDNF-induced aggregation recruits Rho GTPase Rac1 and activates PKC and PI3K/Akt pathway. (A) BDNF-induced aggregation in the presence of Rho GTPase inhibitors (zoledronic acid 50 µM, NSC23766 10 µM, and Y27632 10 µM; n = 5). Repeated-measures ANOVA, P < .0001; inhibition of BDNF with zoledronic acid, P = .0002; inhibition with NSC23766, P = .005; and inhibition with Y27632, P = .53. (B) BDNF-induced phosphorylation of Akt, STAT3, and PLC-γ₂ in the presence of Rho GTPase inhibitors (zoledronic acid 50 µM, NSC23766 10 µM, and Y27632 10 µM). (C) BDNF-induced aggregation in the presence of PI3K and PKC inhibitors (wortmannin 100 nM and BIM-1 10 µM, n = 5). Repeated-measures ANOVA, P < .0001; inhibition of BDNF with BIM-1, P < .0001, inhibition with wortmannin, P < .0001. (D) BDNF-induced phosphorylation of Akt, STAT3, and PLC-γ₂ in the presence of PI3K and PKC inhibitors (wortmannin 100 nM and BIM-1 10 µM). (E) Quantification of BDNF-induced phosphorylation of Akt, STAT3, and PLC-γ₂ in the presence of PI3K and PKC inhibitors (wortmannin 100 nM and BIM-1 10 µM). One-way ANOVA; compared with vehicle: phosphorylation of Akt by BDNF is increased (P < .05), and there is no significant difference in the presence of BIM-1 and wortmannin. Phosphorylation of STAT3 by BDNF is increased (P < .05) with BDNF in the presence of wortmannin (P < .05), and there is no significant difference with BIM-1. Phosphorylation of PLC-γ₂ by BDNF in the presence of wortmannin, P < .05; no significant difference with BDNF alone or in the presence of BIM-1. Density was measured with ImageJ and is expressed as a ratio of density of phosphorylated protein/density of total protein and standardized to the vehicle control. Functional data and phosphoblots are representative of ≥3 independent experiments.