Results 3. although this modification was not statistically significant. Neither G?6976 (2.0 μg i.c.v.) nor hispidin (3.0 μg i.c.v.) significantly altered PKCβI or PKCβII expression at 4 h 3 or 14 d following the last MA dosage (Fig. 2B C). Treatment with MA didn’t induce PKCζ significantly. Furthermore PKCζ pseudosubstrate inhibitor (1.5. or 3.0 μg i.c.v.) didn’t affect PKCζ manifestation at 4 h 3 or14 d following the last MA dosage (Fig. 2D). The duration from the significant MA-induced upsurge in PKCδ manifestation was at least 2 weeks (P < 0.01). Automobile or rottlerin treatment did not affect PKCδ expression. Rottlerin [1.5 or 3.0 μg intracerebroventricularly (i.c.v.)] significantly attenuated MA-induced PKCδ expression in a dose- and time-dependent manner (4 h 3 d Rabbit Polyclonal to AIM2. and 14 d post-MA; Veh + MA vs. 1.5 Acetylcorynoline IC50 μg rottlerin + MA or 3.0 μg rottlerin + MA; P < 0.05 or P < 0.01 respectively; Fig. 2E). 3.2 PKCδ is involved in MA-induced behavioral impairments in mice Because MA-induced behavioral impairment is at least in part related to the dopaminergic degenerative effects of the drug we Acetylcorynoline IC50 measured locomotor activity and rota-rod performance in animals treated with various drug combinations (Fig. 3). Significant decreases in locomotor activity (P < 0.01) and rota-rod performance (P < 0.01) were observed 3 d after the final MA administration. These decreases persisted [locomotor activity (P < 0.05) and rota-rod performance (P < 0.05)] for 14 d after the final MA administration. No Acetylcorynoline IC50 significant changes in locomotor activity and rota-rod Acetylcorynoline IC50 performance were observed in the absence of MA. Vehicle treatment did not affect behavioral impairments induced by MA. The locomotor activity profile consistently paralleled that of rota-rod performance. Treatment with rottlerin a PKCδ inhibitor blocked these changes in both locomotor activity and rota-rod performance (3 d after the final MA; Veh + MA vs. 1.5 or 3.0 μg rottlerin + MA P < 0.05 or P < 0.01 respectively; 14 d after the final MA; Veh + MA vs. 3.0 μg rottlerin + MA P < 0.05). Because the intracerebroventricular (i.c.v.) route is more effective than the oral (p.o.) route in obtaining the neuroprotective effects of rottlerin [38] we used an i.c.v. infusion (Fig. 3A B). Results were comparable to those in MA-treated PKCδ (-/-) mice (data not shown). However treatment with G?6976 (a PKCα and PKCβ inhibitor) hispidin (a PKCβ inhibitor) and PKCζ pseudosubstrate inhibitor showed no significant effect on MA-induced behavioral impairment in mice. 3.3 Effects of rottlerin on MA-induced increases in DA turnover in the striata of PKCδ (+/+) mice: comparison with MA-treated PKCδ (-/-) mice Having shown that MA causes marked changes in PKCδ expression we then examined the involvement of the isozyme in MA toxicity. MA treatment considerably reduced striatal DA amounts [both at 3 d and 14 d post-MA: P < 0.01 vs. saline-treated PKCδ (+/+) mice; Fig. 4A]. MA also considerably reduced 3 4 acidity (DOPAC; both at 3 d and 14 d post-MA: P < 0.05 vs. saline-treated PKCδ (+/+) mice; Fig. 4C) and homovanillic acidity (HVA; both at 3 d and 14 d post-MA: P < 0.05 vs. saline-treated PKCδ (+/+) mice; Fig. 4E). Furthermore MA induced raises in the striatal DA turnover price [(DOPAC + HVA)/DA; both at 3 d and 14 d post-MA: P < 0.01 vs. saline-treated PKCδ (+/+) mice; Fig. 3G]. Automobile or rottlerin treatment didn't alter DA amounts or the DA turnover price. Additionally automobile treatment didn't affect MA-induced dopaminergic (DAergic) adjustments. Rottlerin (1.5 or 3.0 μg i.c.v.) considerably attenuated the MA-induced reduction in DA and upsurge in the DA turnover price inside a dose-dependent way (3 d post-MA; DA: Veh + MA vs. 1.5 or 3.0 μg rottlerin + MA; P < 0.05 or P < 0.01 respectively; DA turnover price: Veh + MA vs. 1.5 or 3.0 μg rottlerin + MA; P < 0.05 or P < 0.01 respectively; 14 d post-MA; DA: Veh + MA vs. 1.5 or 3.0 μg rottlerin + MA; P < 0.05 or P < 0.01 respectively; DA turnover price: Veh + MA vs. 1.5 or 3.0 μg rottlerin +.