Cortical responses to sensory stimuli are modulated by behavioral state. In the primary visual cortex (V1), visual responses of pyramidal neurons increase during locomotion. This response gain was suggested to be mediated through inhibitory neurons, resulting in the disinhibition of pyramidal neurons. Using in vivo two-photon calcium imaging in layers 2/3 and 4 in mouse V1, we reveal that locomotion increases the activity of vasoactive intestinal peptide (VIP), somatostatin (SST) and parvalbumin (PV)-positive interneurons during visual stimulation, challenging the disinhibition model. In darkness, while most VIP and PV neurons remained locomotion responsive, SST and excitatory neurons were largely non-responsive. Context-dependent locomotion responses were found in each cell type, with the highest proportion among SST neurons. These findings establish that modulation of neuronal activity by locomotion is context-dependent and contest the generality of a disinhibitory circuit for gain control of sensory responses by behavioral state. DOI: http://dx.doi.org/10.7554/eLife.14985.001, eLife digest How we perceive what we see depends on the context in which we see it, such as what we are doing at the time. For example, we perceive a park landscape differently when we are running through it than when we are sitting on a park bench. Behavior can also alter neuronal responses in the brain. Indeed, the neurons in the part of the brain that receives information related to vision (known as the visual cortex) respond differently to visual stimuli when an animal is moving compared to when the animal is still. However, while some recent studies revealed that specific types of neurons become more or less responsive during movement, others reported the opposite results. One hypothesis that would explain these contradictory findings would be if the way that behavior, in this case movement, affects neuronal responses also depends on the external context in which the movement happens. Now, Pakan et al. have tested this hypothesis by imaging the activity of different types of neurons in the primary visual cortex of mice that were either running on a treadmill or staying still. The experiments were conducted in two different contexts: in total darkness (in which the mice could not see) and in the presence of display screens (which provided the mice with visual stimulation). Pakan et al. confirmed that running does indeed affect the activity of specific neurons in different ways in different contexts. For example, when the mice received visual stimulation, the three main classes of neurons that send inhibitory signals in the visual cortex became more active during running. However, when the mouse ran in the dark, two of these neuron types became more active during running while the third type of neuron was unresponsive. This finding reveals more about the dynamic nature of inhibitory activity that strongly depends on the animal’s behaviour. It also shows how these neurons influence the excitatory neurons in the visual cortex, which send information to the rest of the brain for further processing towards perception. The next step will be to identify what precise mechanism makes these neurons respond differently in unique contexts, and to tease apart how these movement-dependent signals affect the way animals perceive visual stimuli. DOI: http://dx.doi.org/10.7554/eLife.14985.002