DOI: 10.5061/dryad.x3ffbg7h1
title: The industrial solvent 1,4-Dioxane causes hyperalgesia by targeting capsaicin receptor TRPV1

description: Background: The synthetic chemical 1,4-dioxane is used as industrial
 solvent, food and care product additive. 1,4-Dioxane has been noted to
 influence the nervous system in long-term animal experiments and in
 humans, but the molecular mechanisms underlying its effects on animals
 were not previously known. Results: Here, we report that
 1,4-dioxane potentiates the capsaicin-sensitive transient receptor
 potential (TRP) channel TRPV1, thereby causing hyperalgesia in mouse
 model. This effect was abolished by CRISPR/Cas9-mediated genetic deletion
 of TRPV1 in sensory neurons, but enhanced under inflammatory conditions.
 1,4-Dioxane lowered the temperature threshold for TRPV1 thermal activation
 and potentiated the channel sensitivity to agonistic stimuli. 1,3-dioxane
 and tetrahydrofuran which are structurally related to 1,4-dioxane also
 potentiated TRPV1 activation. The residue M572 in the S4-S5 linker region
 of TRPV1 was found to be crucial for direct activation of the channel by
 1,4-dioxane and its analogues. A single residue mutation M572V abrogated
 the 1,4-dioxane-evoked currents while largely preserving the capsaicin
 responses. Our results further demonstrate that this residue exerts a
 gating effect through hydrophobic interactions and support the existence
 of discrete domains for multimodal gating of TRPV1 channel. Conclusions:
 Our results suggest TRPV1 is a co-receptor for 1,4-dioxane, and that this
 accounts for its ability to dysregulate body nociceptive sensation.
description: Electrophysiological data were
 mainly collected from traditional patch-clamp recordings and
 Ca<sup>2+</sup> imaging. All behavioral experiments with mice
 were conducted in a double-blind manner. Paw-withdrawal latency of thermal
 or mechanical hyperalgesia was determined by a temperature-controlled
 Plantar Test Instrument and von Frey apparatus,
 respectively.
description: This dataset contains nine excel sheets including all data used
 to analyze the effect of the industrial solvent 1,4-Dioxane on thermal
 TRPV1 channel. Each excel sheet corresponds to a figure, and all
 statistical results along with the statistical methods are exhibited in
 the excel sheet. 1) Fig1-1,4-Dioxane ms figsupplement
 data Data used for Figure 1 ‘1,4-Dioxane causes mice
 hyperalgesia via TRPV1 activation’. (1) Experimental data from mice
 obtained using the Radiant Heat and Von Frey assay and was illustrated in
 Fig 1a&b. (2) Data collected for paw volume examination and was
 shown in Fig 1c. (3) Ca<sup>2+</sup> imaging data and
 electrophysiological data obtained in acutely dissociated DRG and TG
 neurons from <i>Trpv1<sup>+/+</sup></i> and
 <i>Trpv1<sup>-/-</sup></i> mice and were
 demonstrated in Fig 1d-m. 2) Fig2-1,4-Dioxane ms
 figsupplement data Electrophysiological data were
 collected from TRPV1-expressing HEK293 cells by single-channel recordings
 or whole-cell recordings and were used to illustrate the gating and
 modulation of TRPV1 channels by 1,4-dioxane as shown in Figure
 2. 3) Fig3-1,4-Dioxane ms figsupplement data
 This data set was obtained from TRPV1-expressing HEK293 cells
 stimulated by heat and was used to analyze the current amplitudes and
 normalized responses as illustrated in Figure 3. 4)
 Fig4-1,4-Dioxane ms figsupplement data Data were
 collected to analyze the activation of TRPV1 under inflammation-related
 conditions in Figure 4. Electrophysiological data were obtained and
 analyzed for Figure 4a-f. Behavior data shown in Figure 4g were collected
 from mice under carrageenan-induced inflammation conditions using the
 Radiant Heat and Von Frey assay. 5) Fig5-1,4-Dioxane ms
 figsupplement data Electrophysiological data obtained
 by whole-cell recordings in HEK293 cells were used for Figure 5 to
 demonstrate the residue M572 within the S4-S5 linker region of TRPV1
 mediates the effect of 1,4-dioxane and its analogues.
 6) Fig6-1,4-Dioxane ms figsupplement data
 Electrophysiological data were collected for Figure 6. Whole-cell
 recordings were performed in HEK 293 cells transiently transfected with
 individual mutation. 7) FigS1-1,4-Dioxane ms
 figsupplement data Electrophysiological data were
 collected for Figure S1 to show effects of 1,4-dioxane on variable TRP
 channels. 8) FigS3-1,4-Dioxane ms figsupplement
 data Electrophysiological data were obtained for Figure
 S3 to illustrate the amplitude of TRPV1 single-channel currents elicited
 by different conditions. 9) FigS4-1,4-Dioxane ms
 figsupplement data Electrophysiological data were
 collected for Figure S4 to exhibit the inhibitory effect of ruthenium red
 (RR) on TRPV1 currents.