Therapeutic inhibition of keratinocyte TRPV3 sensory channel by local anesthetic dyclonine
- State Key Laboratory of Virology, Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Frontier Science Center for Immunology and Metabolism, Wuhan University, China
- School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, China
- Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, China
- State Key Laboratory of Natural Medicines and Jiangsu Provincial Key Laboratory for TCM Evaluation and Translational Development, School of Traditional Chinese Pharmacy, China Pharmaceutical University, China
- Sorbonne Université, Institute of Biology Paris Seine, Neuroscience Paris Seine, CNRS UMR8246, Inserm U1130, France
Figures
Figure 1
Inhibition of transient receptor potential vanilloid-3 (TRPV3) currents by dyclonine (Dyc).
(A) Inhibition of 2-aminoethoxydiphenylborate (2-APB)-evoked currents by Dyc in a representative HEK 293T cell expressing mouse TRPV3. After sensitization by repeated application of 100 μM 2-APB, …
Figure 2
Dyclonine (Dyc) is a potent inhibitor of transient receptor potential vanilloid-3 (TRPV3) channel.
(A) Representative inward current traces from whole-cell voltage-clamp recordings show the inhibitory effects of 10 μM Dyc on TRPV1 (A1), TRPV2 (A2), TRPV3 (A3), TRPM8 (A4), or TRPA1 (A5) channels …
Figure 3
The inhibitory effect of dyclonine on transient receptor potential vanilloid-3 (TRPV3) channel is voltage-independent.
(A) Representative whole-cell currents evoked by voltage steps (inset) together with 40 μM 2-aminoethoxydiphenylborate (2-APB) in the absence and presence of 10, 30 μM dyclonine or 10 μM ruthenium …
Figure 4
Inhibition of heat-activated transientreceptor potential vanilloid-3 (TRPV3) currents by dyclonine.
(A) Sensitization of TRPV3 by heat. Heat-evoked TRPV3 currents in response to repeated temperature jumps. Temperature pulse generated by infrared laser diode irradiation was stepped from room …
Figure 5
Dyclonine (Dyc) rescued cell death caused by expression of overactive transient receptor potential vanilloid-3 (TRPV3) mutant.
(A, B) Effects of Dyc on whole-cell currents recorded from TRPV3 (G573S) and TRPV3 (G573C) expressed in T-Rex 293 cells, showing that Dyc (3 and 10 μM) reversibly inhibited the response to 300 μM …
Figure 6 with 1 supplement
Dyclonine (Dyc) suppresses scratching behavior induced by carvacrol.
(A) Summary of the time courses of neck-scratching behaviors in wild-type transientreceptor potential vanilloid-3 (TRPV3) and TRPV3 knock out (C57BL/6) mice after intradermal injection of 50 μl …
Figure 6—figure supplement 1
Effects of dyclonine on the excitability of mouse dorsal root ganglia (DRG) and trigeminal ganglia (TG) neurons as well as the transient receptor potential vanilloid-3 (TRPV3)-mediated nociceptive behavior.
(A, B) Current-clamp responses of DRG and TG neurons to 500 ms current pulse injection, respectively. Protocol of injected current is shown on the top. (C) Statistics plot showing no significant …
Figure 7
Effects of dyclonine on single-channel properties of transient receptor potential vanilloid-3 (TRPV3).
(A) Single-channel currents of TRPV3 were recorded from inside-out membrane patches of HEK 293T cells at two membrane potentials (± 60 mV) in symmetrical 150 mM NaCl and were low-pass filtered at 2 …
Figure 8 with 2 supplements
Molecular residues involved in dyclonine-transient receptor potential vanilloid-3 (dyclonine-TRPV3) interaction.
(A) Overall view of the mTRPV3-dyclonine complex. Three putative binding modes (BMs) for dyclonine in the pore cavity of mTRPV3 channel (PDB ID code: 6DVZ) are denoted as BMA, BMB, and BMC (please …
Figure 8—figure supplement 1
Residues in the transient receptor potential vanilloid-3 (TRPV3) channel pore for interacting with dyclonine.
(A) Receptor grid for docking generated with 35 Å × 35 Å × 35 Å dimensions. (B) Detailed binding modes (BMs) of dyclonine in BMB and BMC. The putative interaction residues are labeled, and dyclonine …
Figure 8—figure supplement 2
Alignment of the pore-region sequences.
Alignment of the pore sequence of mTRPV3 with other TRP channels, with the identical or similar residues shaded in colors. Meanwhile, differences in amino acids compositions present for different …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Species (Mus musculus) | Trpv3-/- mice | Wang et al., 2021 | PMID:32535744 | C57BL/6J background |
| Cell line (Homo sapiens) | HEK 293T | ATCC | Cat.#:CRL-3216 | |
| Cell line (Homo sapiens) | T-Rex 293 | Thermo Fisher | Cat.#:R71007 | |
| Chemical compound | 2-APB | Sigma-Aldrich | Cat.#:D9754 | TRPV1-3 agonist |
| Chemical compound | Carvacrol | MedChemExpress | Cat.#:499752 | TRPV3 agonist |
| Chemical compound | Menthol | Sigma-Aldrich | Cat.#:M278 | TRPM8 agonist |
| Chemical compound | Capsaicin | MedChemExpress | Cat.#: HY10448 | TRPV1 agonist |
| Chemical compound | AITC | Sigma-Aldrich | Cat.#:377430 | TRPA1 agonist |
| Chemical compound | Ruthenium red | Sigma-Aldrich | Cat.#:R2751 | TRP channel inhibitor |
| Chemical compound | Poly-L-lysinehydrochloride | Sigma-Aldrich | Cat.#:2658 | |
| Chemical compound | MTEST | MedChemExpress | Cat.#:690632554 | |
| Chemical compound | DTT | Sigma-Aldrich | Cat.#:3483123 | |
| Chemical compound | Dyclonine | MedChemExpress | Cat.#:536436 | |
| Software, algorithm | Patchmaster | HEKA Electronics | ||
| Software, algorithm | OriginPro | Originlab.com | ||
| Software, algorithm | Clampfit 10 | Molecular Devices | ||
| Software, algorithm | SigmaPlot 10 | SPSS Science |
