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. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: J Neurochem. 2013 Mar 18;125(5):663–672. doi: 10.1111/jnc.12229

Fig. 3.

Fig. 3

PKCβ inhibition abolishes quinpirole-induced increase in striatal surface DAT expression and DA uptake in mice. (A) Striatal synaptosomes from the PKCβ+/+ and PKCβ−/− mice were incubated for 15 min with vehicle (Veh) or quinpirole (Quin, 10 μM) before biotinylation of surface DAT. Quinpirole increased surface DAT content in PKCβ+/+ mice but not in PKCβ−/− mice. Representative blots for surface DAT biotinylation under various conditions are shown. Surface DAT was calculated as a ratio of surface biotinylated DAT to the total DAT from the lysate. Data were expressed as percent of the Veh treatment in each genotype. Veh values for ratios of biotinylated/total DAT for PKCβ+/+ and PKCβ−/− mice were 1.1 ± 0.1 and 0.7 ± 0.2, respectively. (B) Pretreatment with the PKCβ inhibitor LY379196 (LY, 100 nM, 1 hr) blocked the quinpirole-induced increase in surface DAT content in PKCβ+/+ mice compared to the Veh treatment and had no effect on surface DAT in PKCβ−/− mice. Representative blots for surface DAT biotinylation under various conditions are shown. Data were calculated as percent of the Veh+Veh treatment. (C) Quinpirole induced a significant time-dependent DAT trafficking in PKCβ+/+ mice, There was a significant increase in surface DAT expression upon quinpirole treatment at 1, 5, 15 and 30 min compared to the Veh treatment. By contrast, quinpirole did not increase surface DAT expression at any time point in PKCβ−/− mice (N=13, 6, 6, 9, 3 and 5 for Veh, 1, 5, 15, 30 and 60 min, respectively). *p<0.05, **p<0.01 vs. Sal, post hoc Bonferroni test. (D) Quinpirole (Quin) increased DA uptake in striatal synaptosomes from PKCβ+/+ mice (N=6, Veh vs. Quin, paired t-test, **p<0.01) but not from PKCβ−/− mice. Data were expressed as percent of Veh treatment in each genotype. Veh values for [3H]DA uptake in PKCβ−/− mice were 94% of that in PKCβ+/+ mice, N=6. (E) The PKCβ inhibitor LY379196 (LY, 100 nM) inhibited quinpirole-stimulated DA uptake in striatal synaptosomes from PKCβ+/+ mice (**p<0.01). Data were calculated as percent of Veh+Veh treatment. Error bars represent SEM.