Single-cell transcriptomic analysis of the adult mouse spinal cord reveals fundamental molecular diversity of autonomic and skeletal motor neurons

To characterize the diversity of cholinergic neurons in the spinal cord, we performed single-nucleus RNA sequencing on cholinergic-enriched pools of nuclei from the adult mouse. Enrichment: Nuclei were enriched by fluorescence-activated nuclei sorting (FANS) of GFP+ cholinergic nuclei from reporter mice. Barcoding: Nuclei were loaded into a 10X Chromium device and barcoded via oligo-dT reverse transcription using the V3 chemistry. Library prep: Libraries were prepared by PCR and size-exclusion according to published 10X protocols. Sequencing: Libraries were sequenced using NextSeq 550 high-throughput, 150-cycle kits and NovaSeq 100-cycle kits.