Fig. 5.

AMPH-mediated NET internalization depends on CaMKII and RhoA activation, independently. a Desipramine sensitive ³H-DA uptake in primary LC cultures decreases in response to a 30-min AMPH (10 μM) pretreatment. Co-application of the CaMKII inhibitor, CK59, (100 uM) during the pre-treatment period blocked the effects of AMPH on NET-mediated ³H-DA uptake. b HEK293 cells transiently transfected with DAT or with DAT and EAAT3 were examined for their DA or Glutamate uptake, respectively. Both transporters lost function in response to AMPH pretreatment. CK59 pretreatment, however, only significantly blocked the effects of AMPH on DAT and EAAT3 sensitivity to AMPH remained. c, d HEK293 cells expressing DAT were treated with AMPH which lead to an increase in activated RhoA, as observed previously. The CaMKII inhibitor had no effect on the Rho activation. e GFP-tagged NET internalization was measured by TIRF microscopy. The loss of membrane localized NET in response to AMPH was blocked by both a Rho inhibitor (red) as well as the CaMKII inhibitor, CK59 (green) (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by two-way ANOVA compared to vehicle control. F_{(1, 30)} = 4.436 for a, F_{(3, 62)} = 9.516 for c)