Motor cortex neurons are activated at a variety of timing during a voluntary movement.However, how excitatory and inhibitory neurons in different cortical layers participate in organizing the voluntary movement remains to be known.Here, we obtained juxtacellular recording and multi-neuron recording from the motor cortex of actively behaving rats, and demonstrated temporally and functionally distinct activations of excitatory pyramidal cells and inhibitory fast-spiking (FS) interneurons.Across the cortical layers, pyramidal cells were activated diversely for sequential motor phases (e.g., preparation, initiation, execution).