Single-Cell Transcriptomics of Mouse Lumbar DRG Neurons

Lumbar DRG neurons from adult mice fed on a regular diet (n = 5 animals) and a high-fat diet (n = 5 animals) were isolated and dissociated following protocol established by Zeisel et al., 2018. A high viability single-cell suspension was prepared, and using the 10X Genomics chromium single cell kit v2, about 6000-8000 cells were recovered. Two rounds of the experiment (total n = 10 animals per group) were performed, and downstream cDNA synthesis, library preparation, and sequencing were performed according to the manufacturer's instruction. Illumina runs were demultiplexed and aligned using the 10X Genomics cell ranger pipeline. Dimensionality reduction, clustering, and differential expression of genes were done in Seurat v4.2.^{35 (link)} Raw matrix files for single-cell RNA sequencing were deposited in Dryad (https://doi.org/10.5061/dryad.9s4mw6mm5).