, 2009). Boosting mGluR1s, for example with positive allosteric modulators I-BET151 research buy to remove GluN3-containing NMDARs, may ultimately restore normal synaptic transmission, prevent adaptations in downstream circuits, and stop the development of addiction. C57/BL6 mice (male and female)

and NR3A heterozygotes and knockouts were injected with cocaine (15 mg/kg i.p.) or the same volume of saline as controls. The dose of cocaine that we used did not induce seizures or increase mortality. The study was conducted in accordance with the Institutional Animal Care and Use Committee of the University of Geneva and with permission of the cantonal authorities (Permit No. 1007/3592/2). The majority of electrophysiology recordings were undertaken in young mice (P14–40) to facilitate identification of VTA cells. However,

note that maturation of excitatory transmission in the VTA of mice is complete at P14 (Bellone Cabozantinib purchase et al., 2011) and we have also reported cocaine-evoked plasticity of VTA DA neuron excitatory synapses in adult mice (aged 7 months, Mameli et al., 2009). After animals were sacrificed, 250-μm-thick horizontal midbrain slices containing the VTA were prepared and whole-cell voltage-clamp recordings were made as previously shown (Bellone and Lüscher, 2006). The access resistance was monitored by a hyperpolarizing step of −14 mV with each sweep, every 10 s. The cells were recorded at the access resistance from 10–25 MΩ, and data were excluded when the resistance changed > 20%. Synaptic currents were evoked by stimuli (0.05–0.1 ms) at 0.1 Hz through a stimulating electrode placed rostral to the VTA. The experiments were carried out in the presence of GABAA receptor antagonist picrotoxin (100 μM); the AMPAR EPSCs were pharmacologically isolated by application of the NMDARs antagonist D,

L-APV (100 μM), whereas the NMDA EPSCs were pharmacologically isolated by the application of the AMPARs antagonist NBQX (10 μM). Dipeptidyl peptidase Representative example traces are shown as the average of 20 consecutive EPSCs typically obtained at each potential or, in the case of plasticity protocols, during the last 5 min of the baseline and at least 30 min after the induction of plasticity. The decay time τw of NMDA EPSC was calculated as described previously (Bellone and Nicoll, 2007). The rectification index of AMPARs is the ratio of the chord conductance calculated at negative potential divided by the chord conductance at positive potentials. I-V curves of pharmacologically isolated NMDARs were generated holding the cells at different membrane potential for 5 min each and normalizing EPSCs at 40 mV. Tricine (N-tris(hydroxymethyl)methylglycine, 10 mM) was used to buffer zinc following the relationship [Zn]free = [Zn]added/200 ( Paoletti et al., 1997). All drugs were purchased from Tocris, except Tetanus Toxin that was purchased from Sigma Aldrich.