Skip to main content
. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: Pain. 2021 Jan;162(1):203–218. doi: 10.1097/j.pain.0000000000002007

Figure 1: Single-nucleus RNA sequencing of dynorphin-lineage (pDyn) spinal nuclei reveals 11 distinct clusters.

Figure 1:

(A) The dorsal half of the left lumbar spinal cord (L3-L5) was harvested and homogenized to liberate intact nuclei (1). Fluorescence-activated nuclear sorting (FANS) selectively isolated GFP-labeled nuclei of pDyn cells (2), which were subjected to single-nucleus capture (3) and RNA-sequencing (4) using 10X Chromium Single Cell Gene Expression assays (n = 7,469 nuclei pooled from 16 mice: 8 naïve and 8 with P3 hindpaw incision). (B) Heat map showing Z-score of scaled and log-transformed normalized expression per cell of the top 10 differentially expressed genes in each cluster, ranked by adjusted p-value. (C) UMAP dimensional reduction plot depicting 11 clusters of spinal pDyn nuclei. (D) UMAP plot showing mean expression of neuronal marker genes Snap25, Syn1, and Syp. Expression was robust in all clusters except Glia10. (E) UMAP plot showing mean expression of inhibitory neuron markers Slc32a1, Gad1, Gad2, and Slc6a5. (F) UMAP plot showing mean expression of excitatory marker Slc17a6. In general, inhibitory and excitatory markers segregate into spatially distinct clusters.