Vu0463271

On the day of the experiment, midazolam (Sandoz, Tokyo, Japan) was diluted with 5× saline (final 1 mg/ml) to be administered at 0.02 ml/g of body weight. CLP290 was received from Dr. Yves De Koninck (Institut Universitaire en Santé Mentale de Québec) or purchased (#SML-2172, Sigma-Aldrich, St. Louis, MO, USA). CLP290 was adjusted to a final concentration of 0.66% prior to oral administration; the CLP290 from Dr. De Koninck was dissolved in 10% hydroxypropyl-β-cyclodextrin, whereas the CLP290 from Sigma-Aldrich was suspended in 0.5% carboxy methylcellulose. On the day of the experiment, VU0463271 (#4719/10, Tocris Bioscience, Bristol, UK) was dissolved in 0.75% dimethyl sulfoxide, at a final concentration of 1 mM.

The KCC2 blocker VU0463271(Tocris) (10 µM), the PKC blocker GF-109203X (Cayman) (10 µM), the PKA blocker H89 dihydrochloride (Tocris) (50 µM), The CaMKII specific blocker KN93 (Cayman) (10 µM), the cell-CaMKII penetrating peptide inhibitor, tatCN21 (GL Biochem. Shanghai) (5 µM), the GABA_A blocker BMI Tocris) (20 µM), the AMPA receptors blocker DNQX (Cayman) (0.2–20 µM) and NMDA receptors blocker APV (Sigma) (50 µM) were applied via the medium solution. For experiments were the VU0463271 was applied, each neuron was recorded prior to and after the KCC2 blocker application. Each neuron was exposed 30 minutes to the drug, prior to determining its effect on the IPSPs or IPSCs reversal potential.