Fig. 3: Single-cell transcriptomic profiling of DRGO cellular subtypes.

a, b Immunocytochemistry on DIV 40 DRGOs (a) and relative quantification (b) of TRK A/B/C combination distinguishing the nociceptive, mechanoreceptive, proprioceptive and undefined neuronal subtypes (arrowhead). Mean ± s.d. n = 5 independent experiments, 6-9 organoids/experiment. Scale bar, 10 μm. c Uniform Manifold Approximation and Projection (UMAP) plot displaying multidimensional reduction and clustering of single-cell RNA-Seq data from DIV 80 DRGOs, and pie chart showing relative abundances of the different cell types. SNP, sensory neuron precursors; IP, immature proprioceptors; P, proprioceptors; N, nociceptors; IM, immature mechanoceptors; M, mechanoceptors; S, satellite cells; Sw, Schwann cells; UN, unknown. (d–i) UMAP plots highlighting normalized expression values of NEUROG2 associated to immature sensory neurons (d), NTRK1 associated to immature and mature nociceptors (e), NTRK2 associated to mechanoreceptors (f), NTRK3 associated to both proprioceptors and mechanoreceptors (g), MSX1 associated to satellite cells (h), and MPZ associated to Schwann cells (i). j Heatmap showing normalized expression values of cell lineage-specific genes within the different clusters. k Representative image of a Syn-GFP patched neuron. Scale bar, 50 μm. l Percentage of DRGO neurons with different action potential patterns following a 100 pA current injection and recorded by current-clamp electrophysiological registrations. n = 3 independent experiments, 2 DRGO/experiment. m Percentage of DRGO neurons that exhibit spontaneous excitatory postsynaptic currents in voltage-clamp electrophysiological recording. n = 3 independent experiments, 2 DRGO/experiment. n Percentage of patched DRGO neurons that exhibit specific neuronal subtypes after single-cell-RT-PCR. n = 3 independent experiments, 2 DRGO/experiment.