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. 2019 Jun 13;9:8625. doi: 10.1038/s41598-019-44866-0

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Inhibition of TRPA1 attenuates epidermal nerve Ca²⁺ increases and itch in Spade mice. (a) Ratio of ear scratch counts after to before topical treatment of the Spade ear skin with vehicle or HC-030031. (b) Intravital multiphoton images (16 μm projection depth) of the ear epidermis of Spade Nav1.8-tdTomato/GCaMP3 mice after treatment with vehicle or HC-030031. (c) Fluorescence intensity ratio of GCaMP3 to tdTomato. Shown are the data at the individual positions in the epidermal nerve fibers. Different colors of the data points indicate different mice in each group. (d) The area-normalized number of nerve fibers showing Ca²⁺ increases (fluorescence intensity ratio of GCaMP to tdTomato >0.15) sustained for at least 30 min. (e) Longitudinal analysis of the clinical score of the indicated mouse groups. The data are shown as the mean ± s.e.m. in (a) (n = 5), (d) (n = 3), and (e) (Spade: n = 7–12, TRPA1-deficient Spade: n = 5–7). *p < 0.05, **p < 0.01.