Figure 2.

Blockade of mammalian target of rapamycin complex 1 (mTORC1)/p70 rpS6 kinase (S6K) signaling does not affect haloperidol-induced phosphorylation of ribosomal protein S6 (rpS6). Wild-type mice were treated with vehicle (Veh), haloperidol (Hal; 0.5 mg/kg), or Hal plus rapamycin (5 mg/kg; administered once per day in a volume of 5 ml/kg body weight, starting 3 days before administration of Hal) and perfused after 15 or 60 min. (a) Phospho-Ser235/236-rpS6 (P-235/236-rpS6) immunoreactivity in single confocal sections of the dorsal striata of mice treated with vehicle (Veh–Veh), haloperidol (Veh–Hal), or haloperidol plus rapamycin (Rapamycin–Hal) and perfused 15 min later. Scale bar=30 μm. (b) Quantification of P-235/236-rpS6 immunoreactive neurons in the dorsal striata of mice treated with Veh–Veh, Veh–Hal, or Rapamycin–Hal (^**p<0.001 vs Veh–Veh). (c) Wild-type mice were treated with Veh, Hal (0.5 mg/kg), or Hal plus rapamycin (5 mg/kg; see above) and were killed after 15 min. Representative autoradiogram showing the reduction of phospho-Thr389-S6K immunoreactivity in the striatum of a mouse treated with rapamycin. (d) Wild-type or S6K1 knockout (KO) mice were treated with vehicle or haloperidol (0.5 mg/kg) and killed 15 min later. Striatal levels of P-235/236-rpS6 were determined by western blotting. Top panels show representative autoradiograms obtained using antibodies against P-235/236-rpS6 (upper) and total rpS6 (lower). Bottom panel is a summary of data represented as means±SEM (n=5; ^*p<0.05 vs respective Veh). (e) Representative autoradiogram showing absence of S6K immunoreactivity in the striatum of an S6K1 KO mice compared with a wild-type mouse.