Neural populations can change the computation they perform on very short timescales. Although such flexibility is common, the underlying computational strategies at the population level remain unknown. To address this gap, we examined population responses in motor cortex during reach preparation and movement. We found that there exist exclusive and orthogonal population-level subspaces dedicated to preparatory and movement computations. This orthogonality yielded a reorganization in response correlations: the set of neurons with shared response properties changed completely between preparation and movement. Thus, the same neural population acts, at different times, as two separate circuits with very different properties. This finding is not predicted by existing motor cortical models, which predict overlapping preparation-related and movement-related subspaces. Despite orthogonality, responses in the preparatory subspace were lawfully related to subsequent responses in the movement subspace. These results reveal a population-level strategy for performing separate but linked computations.

SUMMARY Blocking motor cortical output with lesions or pharmacological inactivation has identified movements that require motor cortex. Yet when and how motor cortex influences muscle activity during movement execution remains unresolved. We addressed this ambiguity using measurement and perturbation of motor cortical activity together with electromyography in mice during two forelimb movements that differ in their requirement for cortical involvement. Rapid optogenetic silencing and electrical stimulation indicated that short-latency pathways linking motor cortex with spinal motor neurons are selectively activated during one behavior. Analysis of motor cortical activity revealed a dramatic change between behaviors in the coordination of firing patterns across neurons that could account for this differential influence. Thus, our results suggest that changes in motor cortical output patterns enable a behaviorally-selective engagement of short-latency effector pathways. The model of motor cortical influence implied by our findings helps reconcile previous observations on the function of motor cortex.

Neuroscientists increasingly analyze the joint activity of multi-neuron recordings to identify population-level structure that is believed to be significant and scientifically novel. Claims of significant population structure support hypotheses in many brain areas. However, these claims require first investigating the possibility that the population structure in question is an expected byproduct of simpler features known to exist in data. Classically, this critical examination can be either intuited or addressed with conventional controls. However, these approaches fail when considering population data, raising concerns about the scientific merit of population-level studies. Here we develop a framework to test the novelty of population-level findings against simpler features such as correlations across times, neurons and conditions. We apply this framework to test two recent population findings in prefrontal and motor cortices, providing essential context to those studies. More broadly, the methodologies we introduce provide a general neural population control for many population-level hypotheses.

A time-consuming preparatory stage is hypothesized to precede voluntary movement. A putative neural substrate of motor preparation occurs when a delay separates instruction and execution cues. When readiness is sustained during the delay, sustained neural activity is observed in motor and premotor areas. Yet whether delay-period activity reflects an essential preparatory stage is controversial. In particular, it has remained ambiguous whether delay-period-like activity appears before non-delayed movements. To overcome that ambiguity, we leveraged a recently developed analysis method that parses population responses into putatively preparatory and movement-related components. We examined cortical responses when reaches were initiated after an imposed delay, at a self-chosen time, or reactively with low latency and no delay. Putatively preparatory events were conserved across all contexts. Our findings support the hypothesis that an appropriate preparatory state is consistently achieved before movement onset. However, our results reveal that this process can consume surprisingly little time.