However, in the current paradigm, we also tried to incorporate those relatively immediate behavioral alterations during the early stages (onsets) of place preference
learning, Navitoclax supplier before the drug is metabolically either degraded from the brain. Thus, the IC-CPP training and testing protocol used in this project was slightly different from methods used by Inhibitors,research,lifescience,medical Ricoy and Martinez, 2009 (Ricoy and Martinez 2009). The testing module started immediately after establishing the baseline place preference as defined above in “Preconditioning phase” and following the conditioning as described in “Conditioning phase”. Testing sessions were as follows: Test #1, immediately after conditioning #1; Test #2, immediately after conditioning #2; Test #3, (no treatment), 24 h following conditioning; and, Test #4, (no treatment), a week following conditioning: Note that to test the role of each nucleus within Inhibitors,research,lifescience,medical the hippocampus-VTA loop, testing was repeated according to the module three times. This means one module per brain area tested (experimental design, Fig. 1). Drugs Ringer’s vehicle solution, Ring (Baxter, Deerfield, IL) was used to mimic artificial cerebrospinal fluid. Ringer’s was
composed of (mg/mL): 6 NaCl, 3.1 Sodium lactate, 0.3 KCl, and 0.2 CaCl2. Dextromethamphetamine hydrochloride Inhibitors,research,lifescience,medical (METH, Sigma Chemical Co., St. Louis, MO) was dissolved in Ringer’s at the concentration of 10 μg/μL. Inhibitors,research,lifescience,medical METH was prepared daily. The NMDA receptor noncompetitive antagonist MK-801 (Sigma Chemical Co., St. Louis, MO) was dissolved in freshly prepared METH solution (1:1) at a concentration of 0.1 mM. Histology After the behavioral experiments were completed rats were euthanized using isofluorane gas anesthesia. Brains were then removed Inhibitors,research,lifescience,medical immediately and preserved in methylbutane solution and stored at −80° in a freezer (no perfusion). Coronal sections (100-μm thickness) were mounted onto gelatin-coated slides and subsequently stained with cresyl violet for verification of cannulae tip placements (Fig. S2). Statistical
analysis Raw data were analyzed using one-way GSK-3 analysis of variance (ANOVA) for repeated measurements to determine if the groups differ in preferences before, during, and after conditioning. When significant main effects were detected, a Fisher’s LSD post hoc tests were used for preplanned pairwise comparisons at α = 0.05. To be consistent with our previous report (Ricoy and Martinez 2009), we used time deviation from baseline to represent either place preference (positive values) or place aversion (negative values). Thus, compared to the baseline, a significant increase in time deviation in favor of the drug-paired chambers was interpreted as positive CPP whereas a significant decrease in time deviation was interpreted as place aversion.