Extended Data Fig. 3: NP2B neurons, specificity of Mrgprb4-tdT-Cre recombination and DRG responses.
From: A distributed coding logic for thermosensation and inflammatory pain
(a) Whole mount ISH images comparing methods used to distinguish NP2A and NP2B cells. NP2 cells (orange outlines) can be assigned by expression of Tmem233 and Fxyd2 but not Mrgprd or Nppb/Sst. NP2B cells (green outlines) also express Mlc1 and a lower level of Calca than NP2A cells (magenta outlines); scale bar = 50 µm. In animals where both methods were applied, there was >90 % agreement. (b) Example triple label ISH of a section from an Ai95 mouse carrying an Mrdprb4-tdT-Cre allele showing GCaMP (blue), Mlc1 (green) and tdT (red). Note perfect overlap of Mlc1 and tdT showing that tdT marks NP2B cells; GCaMP is much more broadly expressed. (c) Example image showing alignment of in vivo tdT fluorescence (red) and a spatial map of stimulus evoked activity (green, all stimuli combined) showing that most responding neurons in these mice are not NP2B cells; scale bars (b, b) = 50 µm for merged views. (d) Heatmap showing Ca-transients from lumbar DRG neurons from Ai95 mice (n = 5) carrying the Mrdprb4-tdT-Cre allele divided into 747 GCaMP-only cells and 109 NP2B neurons expressing tdT; changes in GCaMP fluorescence (% ΔF/F) are color-coded as indicated in the scale bar.
