Bay 60 7550

Bay 60-7550 (97% purity) was purchased from Cayman Chemical Company Inc (Chicago, IL), and dissolved in 10% DMSO (Fisher Scientific, Fair Lawn, NJ). The positive control drugs desipramine and diazepam were purchased from Sigma Aldrich (St. Louis, MO). Mice were given Bay 60-7550 at 0.75, 1.5 and 3 mg/kg, desipramine at 10 mg/kg, or diazepam at 1.5 mg/kg, once daily via intraperitoneal injections (i.p.) in a volume of 10 ml/kg body weight 30 min before stress every day. Selection of Bay 60-7550’s working dose was based on our previous studies with minor modifications [17 (link),18 (link)]. Control animals received vehicle only. Behavioral testing was done 24 hrs after the last stress event. Hippocampus and amygdala were dissected from brains immediately after behavioral testing and stored at -80°C until analyses were carried out.

Bay 60-7550 (2- [(3, 4- dimethoxyphenyl)methyl]- 7- [(1R)- 1- hydroxyethyl]- 4- phenylbutyl]- 5-methyl- imidazo[5, 1- f][1, 2, 4]triazin- 4(1H)- one) and KT5823 (2, 3, 9, 10, 11, 12- hexahydro- 10R- methoxy- 2, 9- dimethyl- 1- oxo- 9S, 12R- epoxy- 1H- diindolo[1, 2, 3- fg:3’, 2’, 1’- kl]pyrrolo[3, 4-i][1, 6]benzodiazocine- 10- carboxylic acid, methyl ester) were obtained from Cayman Chemical (Ann Arbor, MI, USA). Myristoylated autocamtide-2-related inhibitory peptide (myr-AIP), MK801 and H89 were purchased from Calbiochem (San Diego, CA, USA). 7-Nitroindazole (7-NI) and Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) were purchased from Sigma Aldrich.
Bay 60-7550 was dissolved in 0.5% dimethyl sulfoxide (DMSO) and was administered via the intraperitoneal route (i.p.). myr-AIP, MK801, KT5823 and H89 were dissolved in artificial cerebrospinal fluid. Bay 60-7550 (1 and 3 mg/kg) or vehicle was given 30 min before stress procedures once per day for 14 days. MK801 (10µM), myr-AIP (20 µM), 7-NI (20 mg/kg), L-NAME (20 mg/kg), KT5823 (20 µM) and H89 (5 µM) were administered 30 min before treatment with Bay 60-7550. Animals were given bilateral microinjections of 2 µl MK801 and myr-AIP (1 µl/side) into the CA1 of the hippocampus. All the behavioral tests were performed 24 h after last drug treatment.