Neuronal ramping and theta power during freely-moving and head-fixed mouse interval timing

An important paradigm to study executive function is interval timing, which requires participants to estimate a temporal interval, often with a motor response. Interval timing translates from rodents to humans and can model neurodegenerative and neuropsychiatric disease. Interval timing also has parallel, time-dependent neurophysiology in rodents and humans, including time-dependent linear changes in neuronal firing rates over a temporal interval (ramping activity) and low-frequency ∼4 Hz “theta” activity evoked by trial start and response. Despite these translational features, an important confound of interval timing is movement and motor preparation, as participants must report their estimates by planning a movement. To address this confound, we first trained a group of 7 mice in a freely-moving interval timing task in which mice had to move across an operant chamber and nosepoke at the correct time to receive food reward. These mice were then trained in a head-fixed version of interval timing in which mice receive liquid reward for holding still for 3 seconds at the correct location, with reward access randomized to probe interval timing behavior. Despite vastly different motor sequences, we found prominent ramping activity in mouse prefrontal cortex ensembles and prefrontal cortical ∼4 Hz activity evoked both by trial start and preceding the timed decision. These data provide evidence that prefrontal neuronal ramping and theta activity is not linked to a specific motor program but rather a feature of the temporal organization of behavior.