In order to investigate the impact of NR1 deletion on the cellular properties of DA neurons, we recorded the

activities of these neurons in both the DA-NR1-KO mice and wild-type control littermates. Movable bundles of 8 tetrodes (32 channels) were implanted into the ventral midbrain, primarily the VTA. The putative DA neurons were identified based on their firing patterns and their sensitivity to dopamine receptor agonist apomorphine (1 mg/kg, i.p.) at the end of each recording session (Figures 3A–3D). A total Ixazomib order of 14 putative DA neurons from 4 mutant mice and 16 from 6 wild-type controls were recorded and analyzed. Phasic-firing activities or bursting was defined as a spike train beginning with an interspike interval (ISI) smaller than 80 ms and terminating with an ISI greater than 160 ms. Compared with the control neurons, phasic-firing

activities were greatly reduced in the DA-NR1-KO neurons. The observed median frequency of phasic firing decreased from 0.78 ± 0.09 Hz in the control DA neurons to 0.36 ± 0.09 Hz in KO DA neurons. (Mann-Whitney U test, p < 0.01) (Figure 3E). A significant reduction was also observed in the percentages of spikes fired EPZ-6438 supplier in phasic activities (34.7% in the controls versus 21.2% in the DA-NR1-KO; Mann-Whitney U test, p < 0.01) (Figure 3F). The total firing rate was also reduced in the mutant DA neurons. This appeared to be correlated with reduced burst set rate (5.18 ± 0.59 Hz, control, versus 3.85 ± 0.38 Hz, KO; r = 0.7719, Mann-Whitney U test, p < 0.01) (Figure 3G). No significant difference was observed in the tonic firing between the mutant and control groups. (4.42 ± 0.44 Hz in control,

versus 3.29 ± 0.36 Hz in KO; Mann-Whitney U test, p > 0.05) (Figure 3H). To further evaluate the response of DA neurons in a learning task, else mice were trained 40 trials per day in a Pavlovian-conditioning paradigm in which a 5 KHz tone that lasted 1 s proceeded immediately before the delivery of a food pellet. DA neurons from both genotypes were able to associate the tone with phasic firing, but the conditioned responses were much weaker in the DA-NR1-KO group (Figure 4A). Although DA-NR1-KO neurons showed increased firing over the days during the training, their responses were significantly reduced compared with the controls on day 1 (19.21 ± 3.24 Hz, control, versus 9.74 ± 0.30 Hz, KO; p < 0.01), day 2 (36.33 ± 4.39 Hz, control, versus 16.43 ± 4.01 Hz, KO; p < 0.01), and day 3 (59.38 ± 3.82 Hz, control, versus 33.88 ± 4.30 Hz, KO; p < 0.01) (Figure 4B). These data suggested that while NMDAR1 deletion did not completely prevent DA neurons from developing conditioned responses (bursting) toward reward-predicting cues, it did, however, greatly lower the robustness of the bursting response, a phenomena that we call DA neuron blunting. To assess habit learning, we first tested the mice in a lever-pressing operant-conditioning task.