Three Types of Cortical Layer 5 Neurons That Differ in Brain-wide Connectivity and Function

•Genetic targeting reveals three types of L5 pyramidal neurons in mouse visual cortex•Morphology, physiology, connectivity, and visual responses of L5 neurons are described•CC and CS neurons receive different inputs from higher visual and association cortex•CS neurons are more direction-selective and prefer faster stimuli than CC neurons

SummaryCortical layer 5 (L5) pyramidal neurons integrate inputs from many sources and distribute outputs to cortical and subcortical structures. Previous studies demonstrate two L5 pyramid types: cortico-cortical (CC) and cortico-subcortical (CS). We characterize connectivity and function of these cell types in mouse primary visual cortex and reveal a new subtype. Unlike previously described L5 CC and CS neurons, this new subtype does not project to striatum [cortico-cortical, non-striatal (CC-NS)] and has distinct morphology, physiology, and visual responses. Monosynaptic rabies tracing reveals that CC neurons preferentially receive input from higher visual areas, while CS neurons receive more input from structures implicated in top-down modulation of brain states. CS neurons are also more direction-selective and prefer faster stimuli than CC neurons. These differences suggest distinct roles as specialized output channels, with CS neurons integrating information and generating responses more relevant to movement control and CC neurons being more important in visual perception.