Trifluoperazine dihydrochloride

Trifluoperazine dihydrochloride (Sigma-Aldrich), SCH23390 (Sigma-Aldrich), and raclopride (Sigma-Aldrich) solution in water was i.p. injected into mice at 0.05 mg/kg, 0.05 mg/kg, and 0.3 mg/kg, respectively. For passive avoidance, trifluoperazine was administered 1 h before or after training. For all other behavioral paradigms, drug was administered 1 h before examination. Control mice were treated similarly but injected with vehicle.
To test the effect of repeated drug administration, trifluoperazine at 0.05 mg/kg was injected once per day for 8 days. One hour after the last injection, mice were examined by behavior tests.

The chemical reagents used in this study are as follows: 100 μM nifedipine (Sigma-Aldrich, St. Louis, MO; N7634); 500 μM gadolinium chloride (Sigma-Aldrich, G7532); 10 μM SKF96365 (Sigma-Aldrich, S7809); 1 M EGTA (Sigma-Aldrich, E4378); 500 μM lanthanum chloride (Sigma-Aldrich, L4131); 2 μM ω-Agatoxin IVA (Tocris, Bristol, UK; 2799); 2 μM SNX 482 (Tocris, 2799); 10 μM ω-Conotoxin GVIA (Alomone Labs, C-300); 10 μM U0126 (Sigma-Aldrich, U120); 50 mM potassium chloride (Sigma-Aldrich, P3911); 100 nM AP24534 (Tocris, 4274); 25 μM trifluoperazine dihydrochloride (Sigma Aldrich, T8516).
The following plasmids were purchased from Addgene: pGP-CMV-GCaMP6s was a gift from Douglas Kim (Addgene plasmid # 40753)^{33 (link)}; pGP-CMV-GCaMP6s-CAAX was a gift from Tobias Meyer (Addgene plasmid # 52228)^{34 (link)}; AAV-CAG-GFP was a gift from Karel Svoboda (Addgene plasmid # 28014)^{35 (link)}. The generation procedures for ERK1-dTomato construct were described previously^{36 (link)}. AAV-CAG-GCaMP6s-CAAX was cloned by replacing the GFP sequence in the AAV-CAG-GFP vector with GCaMP6s-CAAX PCR amplicon flanked BamHI/HindIII restriction sites. pHelper, and pAAV-RC1 plasmids were purchased from Cell Biolabs. RaichuEV-HRas FRET biosensor was kindly gifted from Dr. M. Matsuda (Kyoto University).

Tag-lite labeling medium (LABMED), SNAP-Lumi4-Tb, and the PPHT ((±)-2-(n-phenethyl-n-propyl)amino-5-hydroxytetralin hydrochloride;1-Naphthalenol,5,6,7,8-tetrahydro-6-[(2-phenylethyl)propylamino]) derivative labeled with a red fluorescent probe (PPHT-red) was obtained from Cisbio Bioassays (Bagnols-sur-Cèze, France). Ninety-six-well polypropylene plates (Corning) were purchased from Fisher Scientific UK (Loughborough, UK) and 384-well optiplate plates were purchased from PerkinElmer (Beaconsfield, UK). GppNHp, risperidone, chlorpromazine hydrochloride, quetiapine hemifumarate, ziprasidone hydrochloride monohydrate, zotepine, sertindole, thioridazine hydrochloride, fluphenazine dihydrochloride, molindone hydrochloride, loxapine succinate, perphenazine, trifluoperazine dihydrochloride, spiperone, (−)-sulpiride, droperidol, and (+)-butaclamol used in competition assays were obtained from Sigma-Aldrich (Poole, UK). Olanzapine, nemonapride, remoxipride hydrochloride, flupenthixol dihydrochloride, paliperidone, amisulpride, melperone hydrochloride, clozapine, raclopride, domperidone, asenapine maleate and haloperidol hydrochloride used for competition assays were obtained from Tocris Bioscience (Avonmouth, Bristol).