Embryonic Rat TRPV1+ and IB4+ DRG FACS-sorted neurons. Rattus norvegicus strain:Sprague-Dawley

Cells were loaded with calcium sensitive dyes and stimulated with resiniferatoxin (RTX). The fluorescent signal after RTX-induced calcium influx is detectable by 30s, and lasts at least 15 minutes (A). This RTX-sensitive signal was used for cell-sorting. Dissociated cells were sorted for live vs. dead cells using DAPI (B) and live cells were passed through a second gate based on RTX-induced calcium influx and IB4 fluorescence (C). RTX responsive cells were subdivided by size into small and medium populations. After sorting, cells were sequenced, and differential expression was tabulated for RTX-responsive small cells (TRPV1+) vs. IB4+ cells (D, E). Differential genes were identified, and representative highly expressed marker genes are shown for both the eTRPV1+ (F), and the eIB4+ (G) populations (n = 4). In contrast to a previous dataset using microarray to characterize the adult IB4+ neuronal population (3), the present eIB4+ dataset contains mostly non-neural cells. Examples of highly expressed marker genes for each population are shown in Figure 1E-F. The eTRPV1+ population is highly enriched for Trpv1, as well as several voltage-gated sodium channel genes, and other markers of nociceptive neuronal progenitors (Figure 1E). Because of its embryonic status, the eIB4+ population contains many markers of blood and vasculature such as the hemoglobin genes, and the vascular K+ channel Kcnj8.