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. 2020 Sep 3;9:e61103. doi: 10.7554/eLife.61103

Figure 4. Increased TRPM3 activity in peripheral sensory nerve endings during inflammation.

(A) Normalized fluorescence at baseline (before the first stimulus), and upon stimulation with TRPM3, TRPA1 and TRPV1 agonists of the ipsi- and contralateral skin of a CFA-treated mouse. Scale bar is 50 μm. Boxed areas, magnified on the right, illustrate automatically detected ROIs. (B) GCaMP3 fluorescence, expressed as ΔF/F0, for the ROIs indicated in panel (A). (C-E) Responsive areas to the indicated agonists in the contralateral (black) and ipsilateral (red) skin. The paired Wilcoxon Signed Rank Test was used for a paired comparison of the responsive area in the ipsi- and contralateral paw skin of 11 mice, measured in the absence of isosakuranetin. The ipsi- and contralateral skin of another set of 6 mice was compared following pre-incubation with isosakuranetin (20 μM; grey background). Since the control and isosakuranetin-treated skin preparations originate from different mice, and considering the substantial inter-animal variability in skin thickness and innervation, a full statistical comparison between these data sets was not performed. (F) Percentage of the total imaged area responding to the indicated (combinations of) agonists in the contra- and ipsilateral paws, and following isosakuranetin pre-incubation. Further statistical comparison is provided in Figure 4—figure supplement 1.

Figure 4—source data 1. Raw values used for plots in Figure 4.

Figure 4.

Figure 4—figure supplement 1. Inflammation-induced increases in sensory nerve endings that respond to multiple TRP agonists.

Figure 4—figure supplement 1.

(A-C) Responsive areas that were identified based on their response to TRPM3 (A), TRPA1 (B) or TRPV1 (C) Agonism, and that also responded to at least one other TRP channel agonist. Individual dots and interconnecting full lines represent values of the contralateral and ipsilateral paw of individual mice (N = 11). Mice where the responsive area was increased in the inflamed paw compared to the control paw are indicated in red, those where there was a decrease in green. Solid black circles represent the mean and error bars the 95% confidence interval. Statistical comparisons were made using the paired Wilcoxon Signed Rank Test. The upper symbol (star) represents a highly responsive inflamed skin with actual responsive areas of 7.69% (TRPM3 with TRPA1 and/or TRPV1), 8.23% (TRPA1 with TRPM3 and/or TRPV1) and 8.62% (TRPV1 with TRPM3 and/or TRPA1).