Dissection of Cortical Microcircuits by Single-Neuron Stimulation In Vivo

A fundamental process underlying all brain functions is the propagation of spiking activity in networks of excitatory and inhibitory neurons. In the neocortex, although functional connections between pairs of neurons have been studied extensively in brain slices, they remain poorly characterized in vivo, where the high background activity, global brain states, and neuromodulation can powerfully influence synaptic transmission. To understand how spikes are transmitted in cortical circuits in vivo, we used two-photon calcium imaging to monitor ensemble activity and targeted patching to stimulate a single neuron in mouse visual cortex. Burst spiking of a single pyramidal neuron can drive spiking activity in both excitatory and inhibitory neurons within a ∼100 μm radius. For inhibitory neurons, ∼30% of the somatostatin interneurons fire reliably in response to a presynaptic burst of ≥5 spikes. In contrast, parvalbumin interneurons showed no detectable responses to single-neuron stimulation, but their spiking is highly correlated with the local network activity. Our results demonstrate the feasibility of mapping functional connectivity at cellular resolution in vivo and reveal distinct operations of two major inhibitory circuits, one detecting single-neuron spike bursts and the other reflecting distributed network activity. ► Functional connectivity between cortical neurons was mapped in vivo ► Single-neuron burst spiking drives subset of SOM and pyramidal neurons to fire ► PV interneurons are less sensitive to single-pyramidal neuron stimulation ► Activities of PV interneurons and the local network are highly correlated