Effects of optogenetic inhibition of a small fraction of parvalbumin-positive interneurons on the representation of sensory stimuli in mouse barrel cortex.

Inhibitory interneurons play central roles in the modulation of spontaneous network activity and in processing of neuronal information. In sensory neocortical areas, parvalbumin-positive (PV+) GABAergic interneurons control the representation and processing of peripheral sensory inputs. We studied the functional role of PV+ interneurons in the barrel cortex of anesthetized adult PVCre mice by combining extracellular multi-electrode recordings with optogenetic silencing of a small fraction of PV+ interneurons. In all cortical layers, optogenetic inhibition caused an increase in spontaneous network activity from theta to gamma frequencies. The spatio-temporal representation of sensory inputs was studied by stimulating one or two whiskers at different intervals and analyzing the resulting local field potential (LFP) and single unit (SU) response. Silencing PV+ interneurons caused an increase in LFP response to sensory stimulation and a decrease in temporal discrimination of consecutive whisker deflections. The combined effect of whisker deflection and optogenetic inhibition was highly similar to the linear sum of the individual effects of these two manipulations. SU recordings revealed that optogenetic silencing reduced stimulus detectability by increasing stimulus-evoked firing rate by a constant offset, suggesting that PV+ interneurons improve signal-to-noise ratio by reducing ongoing spiking activity, thereby sharpening the spatio-temporal representation of sensory stimuli.

Pipeline for 2-photon all-optical physiology in mouse: From viral titration and optical window implantation to binarization of calcium transients.

2-photon all-optical physiology combines in vivo 2-photon calcium imaging and optogenetics, which enables both the read out and manipulation of neuronal microcircuits with single-cell resolution. Here, we describe a protocol for achieving optimized co-expression of calcium indicator and opsin. To enable longitudinal designs, we introduce a template for virus injection and chronic window implantation. We also highlight key aspects of performing 2-photon imaging and suggest an analysis algorithm for the binarization of putatively action-potential (AP)-related calcium transients. For complete details on the use and execution of this protocol, please refer to Fu et al. (2021).