Revealing a compulsive phenotype in cholinergic M₄^-/- mice depends on the inter-trial interval initiation settings in...

Muscarinic acetylcholine receptor 4 (M₄) modulates dopaminergic neurotransmission and is a target for novel treatments of schizophrenia, cognitive deficits, and addiction. Impulsive and compulsive behaviors are key traits of addiction, yet the importance of M₄ receptor signaling to these traits is poorly understood. We investigated impulsive action and compulsivity by measuring premature and perseverative responses in the five choice serial reaction time task (5CSRTT). Furthermore, we hypothesized that inter-trial interval (ITI) initiation settings affected training durations and test performances in these experiments.

M₄^-/- and wildtype mice were trained and tested on two versions of the 5CSRTT with different ITI initiation settings. One setting, the head-in condition, allowed the ITI to start while the mouse's head remained in the reward receptacle (magazine). The other setting, the head-out condition, required the mouse to remove its head from the magazine to initiate the ITI.

We did not observe differences in premature or perseverative responses in M₄^-/- mice in either condition, but found evidence of reward-related compulsive behavior in M₄^-/- mice. In the head-in condition, M₄^-/- mice were slower to acquire the 5CSRTT, had more omissions, and had longer correct response latencies than wildtype mice. In the head-out condition, genotypes did not differ in training, but M₄^-/- mice showed small decreases in accuracy. Our findings demonstrate that ITI initiation settings contribute to different training durations and tested behaviors in M₄^-/- mice, suggesting ITI initiation settings are an important consideration for the general use of the 5CSRTT.