Single cell transcriptome analysis of proprioceptive muscle afferents

We report on the single cell RNA sequencing of genetically identified adult, neonatal, and embryonic proprioceptors. High depth sequencing data was acquired for all developmental time points using plate-seq. For adult proprioceptors, bioinformatics analysis identified five molecularly distinct subsets. At least two of these subsets have been validated to correspond to group Ia muscle spindle afferents and group Ib GTO afferents, respectively. Proprioceptors were labeled using intersectional genetic tools and fluorescently labeled neurons were isolated using fluorescence activated cell sorting (FACS). Neurons were directly deposited in lysis buffer in 96-well plates and subjected to RNA isolation, cDNA generation, library preparation and single cell sequencing. Single proprioceptor transcriptomes were filtered for high quality cells (>2000 UMI's), and limited satellite cell contamination (<10% Apoe).