Journal: eLife
Article Title: Keratinocytes mediate innocuous and noxious touch via ATP-P2X4 signaling
doi: 10.7554/elife.31684
Figure Lengend Snippet: Figure 6. Pharmacological inhibition of peripheral P2X4 and not P2X2 and P2X3 receptors causes decreased mechanical responses in vivo. (A) Naive C57BL/6J mice were injected with either 5-BDBD (selective P2X4 antagonist) or vehicle, 1 hr prior to behavioral experiments. Animals injected with 5-BDBD had significantly higher paw withdrawal thresholds than their vehicle controls (*p=0.0190), Mann-Whitney U-test. (B) Animals injected with 5-BDBD showed fewer responses to the 3.61 mN stimulus than the vehicle control (**p=0.0033), Mann-Whitney U-test. (C) Naive C57BL/6J mice were injected with either vehicle, low NF 110 (500 nM) or high NF 110 (5 mM) 1 hr prior to behavior assays. Neither the low nor the high concentration of NF 110 had an effect on the paw withdrawal thresholds of the animals (n.s. p=0.9946), Kruskal-Wallis test. (D) Injection of low- or high concentration NF 110 had no effect on the percent response to a suprathreshold 3.61 mN stimulus (n.s. p=0.6083), Kruskal-Wallis test. (E) 5-BDBD or vehicle was injected 1 hr prior to the behavior assays and 20 min prior to the assays animals were also injected with 900 mM ivermectin or vehicle. Ivermectin with vehicle injection decreased the paw withdrawal thresholds significantly as compared to the ivermectin and 5-BDBD combined injection (****p<0.0001), two-way ANOVA, Tukey post-hoc. (F) Ivermectin injection with 5-BDBD caused significantly fewer responses than when ivermectin was injected with the vehicle control (****p<0.0001), two-way ANOVA, Tukey post-hoc. Furthermore, ivermectin with 5-BDBD was not significantly different from the 5-BDBD with vehicle control (n. s. p=0.9989), two-way ANOVA, Tukey post-hoc. Data are represented as mean ± SEM. The experimenter was blinded to compound treatment. See also Figure 6—figure supplement 1. DOI: https://doi.org/10.7554/eLife.31684.014 The following figure supplement is available for figure 6:
Article Snippet: No. 9000-95-7 Cat#A6237 Chemical compound, drug (5-BDBD (5-(3-Bromophenyl) -1,3-dihydro-2H-benzofuro[3,2-e] -1,4-diazepin-2-one)) 5-BDBD Tocris Bioscience CAS 768404-03-1 Cat# 3579 Chemical compound, drug (ivermectin) IVM or ivermectin Tocris Bioscience CAS 70288-86-7 Cat#1260 chemical compound, drug (dimethyl sulfoxide) DMSO Sigma-Aldrich Cat # 8418-100mL Chemical compound, drug (NF 110) NF 110 Tocris Bioscience CAS 111150-22-2 Cat#2548 Software, algorithm (Pulse) HEKA Electronics http://www.heka.com/ downloads/downloads_main. html#down_patchmaster Software, algorithm (FitMaster) HEKA Electronics http://www.heka.com/ downloads/downloads_main. html#down_fitmaster Software, algorithm (Origin 5.0) Origin Lab http://originlab.com Software, algorithm (Graphpad Prism 7) Graphpad https://graphpad.com/ scientific-software/prism/ Software, algorithm (ANY-maze) ANY-maze http://www.anymaze.co.uk/ index.htm Software, algorithm (LabChart) ADInstruments https://www.adinstruments. com/products/labchart Software, algorithm (DY2000 Multi-channel Potentiostat) DigiIvy http://www.digi-ivy.com/ dy2000.html Other (490 nm LED) 490 nm Thorlabs inc Cat# M490L2 Other (590 nm LED) 590 nm Thorlabs inc Cat# M590L2 Other (Compact T-Cube LED driver) Thorlabs inc CAT# LEDD1B Other (473 nm Laser) 473 nm Laserglow Cat# R471005GX Other (589 nm Laser) 589 nm Laserglow Cat# R581005GX Other (595 nm LED strip) 596 nm LED floor Environmental Lights Cat# amber3528-120-10-reel Other (460 nm LED strip) 460 nm LED floor Environmental Lights Cat# blue3528-120-10-reel Other (BAT-12 Microprobe Thermometer) Physitemp Cat# BAT-12 Continued on next page Moehring et al. eLife 2018;7:e31684.
Techniques: Inhibition, In Vivo, Injection, MANN-WHITNEY, Control, Concentration Assay
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