An automated data extraction and classification pipeline to identify a novel type of neuron within the dorsal striatum based on single-cell patch clamp and confocal imaging data.

Abstract

We employed electrophysiological and fluorescence imaging techniques to describe the characteristics of a novel type of neuron discovered in the mouse dorsal striatum. Transgenic mice that express YFP-tagged channelrhodopsin-2 (ChR2) in neurons driven by the promoter for tyrosine hydroxylase (TH) were used and the intrinsic electrical properties of YFP-positive neurons in the dorsal striatum of these mice were characterized using whole-cell patch clamping in acute brain slices. Passive membrane properties - such as membrane capacitance, resting membrane potential and input resistance -and action potential properties- such as amplitude, kinetics and adaptation - were extracted from raw data files. Filling these neurons with neurobiotin enabled visualization of neuronal morphology via immunohistochemical labeling with streptavidin-conjugated fluorophore. Subsequent two-photon imaging allowed analyses of morphological properties such as somaticsize, dendritic branching (Sholl analysis) and density of dendritic spines. Unbiased analyses and hierarchical clustering of both morphological and functional data allowed us to identify a previously undescribed type of striatal neuron with unique properties. To facilitate identification of this new cell type, an end-to-end automated electrophysiology pipeline was developed that extracts relevant parameters and determines striatal neuron identity using neural-network based classifiers. These data and the software tool will permit other investigators to identify this novel type of neuron in their studiesand thereby better understand theroles thatthese neuronsplay in dorsal striatum circuitry.