Rats were anesthetized with chloral hydrate (400 mg/kg, i.p.), and their heads were set in a stereotaxic apparatus (David Alectinib Kopft Instruments, USA). A bilateral double cannula (2 mm long, 26 gauge; Plastics One Inc., VA, USA) was implanted at the confluence of the Cg1, Cg2, and Cg3 areas of the mPFC, according to the following parameters relative to bregma: + 2.5 mm AP, ± 1 mm L,
and − 2.0 mm V ( Paxinos and Watson, 1986). The double guide cannula was anchored to the skull with dental acrylic and four small screws and was protected with a double dummy cannula and a dust cap. Each rat was given a 5-day recovery period after surgery. Animals were then retrained Selleck BGB324 in the radial arm maze to return them to their pre-surgery baseline performance. An alcohol (100 vol%) stock solution of Δ9-THC (National Institute on Drug Abuse, Rockville, MD, USA) was stored at 4 °C. For the experimental procedures, the alcohol was evaporated, and the residue was resuspended in a vehicle solution (VEH) composed of emulphor (30%) and dimethylsulfoxide at final concentrations of 64, 200, and 360 mg/ml. The volume injected was 0.5 μl, resulting in a final intracortical (IC) administration of 32, 100, and 180 μg Δ9-THC. Dopaminergic drugs were
purchased from Sigma-Aldrich (MO, USA). The antagonist SCH 23390 [R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1 H-3-benzazepine hydrochloride] was diluted in distilled water to 2 mg/ml, and 0.5 μl were injected, resulting in a final IC administration of 1 μg. The antagonist CZP [8-chloro-11-(4-methylpiperazin-1-yl)-5 H-dibenzo[b,e][1,4]diazepine] was diluted in 0.05 N HCl to 6.4 mg/ml, and 0.5 μl were injected, PRKD3 resulting in a final IC administration of 3.2 μg. These doses of SCH and CZP were selected because previous studies in our lab had shown that they have no effect alone on radial maze performance. For control solutions, VEH alone and either saline (SAL) or HCl were administered.
To administer drugs IC to the mPFC, the dust cap and dummy cannula were removed, and a stainless steel, double internal cannula (33 gauge; Plastics One Inc.) extending to 2.7 mm was lowered through the double guide cannula to the sites of infusion. A double cannula connector (Plastics One Inc.) connected the double internal cannula to two 10-μl Hamilton syringes. A volume of 0.5 μl was delivered to each side of the mPFC over 90 s through a microprocessor syringe pump (Series 100; Stoelting, IL, USA). The experimenter gently handled subjects while the drug was administered. A 90-s diffusion period was allowed before removal of the internal cannula and replacement of the dummy cannula and the dust cap.