Deschloroclozapine dihydrochloride dcz

Clozapine-N-oxide dihydrochloride (CNO; Hello Bio, Princeton, NJ, USA) was dissolved in sterile, 0.9% saline to a concentration of 0.30 mg/ml and injected at 10 ml/kg, i.p., 30 min before the beginning of behavioral experiments for a dose of 3 mg/kg. For c-fos studies, CNO was injected 2 hours before perfusion and brain extraction. Deschloroclozapine dihydrochloride (DCZ; Hello Bio) was dissolved in sterile, 0.9% saline to a concentration of 10 μg/ml and injected at 10 ml/kg, i.p., 20 min before the beginning of behavioral experiments for a final dose of 100 μg/kg, which was previously found to induce neural activity in hM3Dq-expressing neurons in mice and monkeys^{17 (link)}.

Ethanol-CTA was conducted across three days, with each daily session separated by 24 hours. All sessions were conducted in the home cage. Mice were habituated to intraperitoneal (i.p.) injections for three days prior to the start of habituation. On day 1 (habituation), mice were given 30 minutes of access to a 0.5% saccharin solution. On day 2 (conditioning), mice were again given 30 minutes of access to a 0.5% saccharin solution, immediately followed by an i.p. injection of vehicle (saline) or 2g/kg EtOH. On day 3 (retrieval), mice were again given 30 minutes of access to 0.5% saccharin. For DREADD experiments, ethanol-CTA was conducted as described, with the addition of a .1mg/kg i.p. injection (Nagai et al., 2020 (link)) of the DREADD actuator Deschloroclozapine dihydrochloride (DCZ, Hello Bio) given 30 minutes prior to saccharin access on retrieval day. Bottles were weighed prior to and after each saccharin presentation session.