trpv3 channel antagonist trpv3 74a  (Tocris)

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) TRPV3 immunostaining (green) in mouse primary somatosensory (S1) cortex, striatum, thalamus, and hippocampus, with DAPI counterstain (blue). Top row left: Postnatal day (P)7; bottom row left: P14; bottom row right: P21. Top row right: Sections incubated with TRPV3 antibody plus TRPV3 blocking peptide in S1 cortex, striatum, thalamus, and hippocampus (green) with DAPI counterstain (blue). Remaining TRPV3 staining is shown in green with DAPI in blue. ( B ) TRPV3 and TRPV4 immunostaining in S1 cortex, striatum, thalamus, and hippocampus at P14. Top row left: TRPV3 (green) and TRPV4 (brick red). Top row right: TRPV3 (green) and TRPV4 (brick red) with DAPI counterstain (blue). Bottom row left: TRPV4 immunostaining in S1BF (inset from top row left) at 10x. Bottom row right: TRPV3 immunostaining in S1BF (inset from top row left) at 10x.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Immunostaining, Incubation, Blocking Assay, Staining

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording whole-cell TRPV3 currents at 30°C (black), 36°C (gray), and 39°C (red) in cortical excitatory pyramidal neurons (PNs) with bath application of camphor (5 mM), a TPRV3 agonist. ( B ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black), 36°C (gray) and 39°C (red) in wild-type (WT) mice: 11 cells from four mice. ( C ) Scatter dot plots of the current density-voltage measurements. ( D ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black) in the presence of camphor (5 mM), a TPRV3 agonist, or camphor (5 mM) + TRPV3 blocker (Forsythoside B, 50 µM) (blue). ( E ) Same as ( D ) but for 36°C. ( F ) Same as ( D ) but for 39°C. ( G ) Current density-voltage (I–V) plot showing the net TRPV3 current (opener – (opener+blocker) condition). In B - F , statistical significance was assessed using a two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques:

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potential and spiking in an excitatory cortical pyramidal neuron (PN) with an intracellular blocker of TRPV3 channels (Forsythoside B, 50 µM) (left) or TRPV4 channels (RN1734, 10 µM) (right) at just-subthreshold V m at 30°C, 36°C, and 39°C in mouse S1 cortex. ( B ) Percentages of cell types obtained from experiment in A . ( C ) Evoked spikes in L2/3 cortical PNs during temperature elevations to 30°C, 36°C, and 39°C under three conditions: no blockers, TRPV3 blocker (Forsythoside B, 50 µM), or TRPV4 blocker (RN1734, 10 µM). ( D ) Correlation between post-synaptic potential (PSP) peak and spike threshold (ST). r =Pearson correlation coefficient with Deming linear regression. ( E ) Same as ( C ) for the L4-evoked late PSP peak. ( F ) Same as ( C ) for input resistance (R in ). Each data point in C – F represents an individual cell. Data were collected from 26 cells in 7 animals for the TRPV3 blocker, 24 cells in 6 animals for the TRPV4 blocker, and 37 cells in 14 animals for the no-block condition. Mean ± SEM is shown in C , E , and F . Statistical significance was assessed using one- or two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). In D , correlations were evaluated using Pearson’s r with Deming linear regression.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Blocking Assay

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potentials and spiking in excitatory cortical pyramidal neurons (PNs) at just-subthreshold Vm at 30°C, 36°C, and 39°C in mouse S1 cortex of wild-type ( Trpv3 +/+ ) and Trpv3 knockout ( Trpv3 -/- ) mice. ( B ) Depolarization required to reach spike threshold (ST) in Trpv3 +/+ and Trpv3 -/- mice at 30°C, 36°C, and 39°C. ( C ) Same as ( B ) but for input resistance (R in ). ( D ) Same as ( B ) but for number of spikes. ( E ) Same as ( B ) but for post-synaptic potential (PSP). ( F ) Setup for recording mouse body temperature (T b ) at room temperature and during fever-range and higher, using an implanted transponder for non-invasive measurement, with exposure to infrared light. ( G ) Time to loss of postural control (LPC), defined as collapse and failure to maintain upright posture, in wild-type ( Trpv3 +/+ ), heterozygous ( Trpv3 +/- ), and Trpv3 knockout ( Trpv3 -/- ) mice. ( H ) Same as in ( G ) but showing T b at seizure onset. ( I ) Same as in ( G ) but for the time from LPC to seizure onset. Each data point in B – E represents an individual cell (three animals per genotype). Statistical significance was assessed using two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). Each data point in G – I represents an individual animal. Statistical significance was assessed using one-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Knock-Out, Control

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) In cortical L2/3 pyramidal neurons (PNs) with synaptically evoked spiking, gradual increases in brain slice temperature from 30 °C to 36°C to 39°C result in four possible outcomes: neurons remain inactive, continue spiking (STAY), stop spiking, or initiate spiking. To spike, PNs must reach the spike threshold (ST), defined as the minimal V m that elicits an action potential, which requires sufficient depolarization via the post-synaptic potential (PSP). STAY neurons consistently reach ST and continue spiking. Neurons that stop spiking fall below the level of depolarization required to reach ST, while neurons that initiate spiking achieve sufficient depolarization to newly reach ST. ( B ) STAY neurons may contain unique ion channels, such as TRPV3. TRPV3 channels are highly permeable to Ca² + ions, and Ca² + influx contributes to PN depolarization. The presence of TRPV3 facilitates greater ion entry, enabling depolarization sufficient to reach ST and sustain spiking, whereas its absence reduces depolarization to levels insufficient for spiking.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Slice Preparation

Journal: Immunity & Inflammation

Article Title: Fragrant TRPV3 agonists act as titratable organic adjuvants to amplify antigen-specific IgG response

doi: 10.1007/s44466-026-00033-5

Figure Lengend Snippet: Fragrant TRPV3 agonists enhance humoral responses and rapidly induce cytokine transcripts in skin. a , b OVA-specific IgG at days 7 and 14 following local co-immunization with OVA (50 μg) and Carvacrol or Camphor (4–100 μg) ( n = 3, mean ± SEM). c , d Expression profiles from mice footpad tissue 1 h after Carvacrol or Camphor injection in comparison to PBS ( n = 3, mean ± SEM). e – g Transcription levels of CCL20, IL-6 and TNF-α in mice footpad at indicated time points after Carvacrol injection ( n = 3, mean ± SEM). h Relative changes in [Ca 2+ ] i over time in A431 cells loaded with the calcium indicator Fluo-4 AM and treated with 300 μM Carvacrol or 2 mM Camphor (black bar) were monitored by flow cytometry. Quantification of peak [Ca 2+ ] i after Carvacrol and Camphor stimulation ( n = 3, mean ± SD). i Transcription levels of cytokines induced with Carvacrol or Camphor at different concentrations for 1 h in A431 cells. ( n = 3, mean ± SD). j , k Addition of CaCl 2 to PBS buffer rescued [Ca 2+ ] i in A431 cells treated with 300 μM Carvacrol or 2 mM Camphor. Quantification of peak [Ca 2+ ] i . ( n = 3, mean ± SD). l , m Relative changes in [Ca 2+ ] i over time in WT and TRPV3-KO A431 cells loaded with the calcium indicator Fluo-4 AM and treated with 300 μM Carvacrol or 2 mM Camphor. Quantification of peak [Ca 2+ ] i . ( n = 3, mean ± SD). n - p Cytokine expression measured by qPCR in A431 cells after Carvacrol (300 μM) with indicated inhibitors ( n = 3, mean ± SD). q , r Inhibitory effects of RR and CsA on Carvacrol and Camphor-enhanced expression of OVA-specific antibody ( n = 5, mean ± SEM). Statistics computed by two-way ANOVA ( a , b , i , n - r ), one-way ANOVA ( e – g ), unpaired two-tailed t-test ( j - m ). ns, not significant. Data are representative of at least three independent experiments

Article Snippet: The baseline expression data of TRPV3 in human cell types are obtained from Human Protein Atlas, www.proteinatlas.org [ , ].

Techniques: Expressing, Injection, Comparison, Flow Cytometry, Two Tailed Test

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) TRPV3 immunostaining (green) in mouse primary somatosensory (S1) cortex, striatum, thalamus, and hippocampus, with DAPI counterstain (blue). Top row left: Postnatal day (P)7; bottom row left: P14; bottom row right: P21. Top row right: Sections incubated with TRPV3 antibody plus TRPV3 blocking peptide in S1 cortex, striatum, thalamus, and hippocampus (green) with DAPI counterstain (blue). Remaining TRPV3 staining is shown in green with DAPI in blue. ( B ) TRPV3 and TRPV4 immunostaining in S1 cortex, striatum, thalamus, and hippocampus at P14. Top row left: TRPV3 (green) and TRPV4 (brick red). Top row right: TRPV3 (green) and TRPV4 (brick red) with DAPI counterstain (blue). Bottom row left: TRPV4 immunostaining in S1BF (inset from top row left) at 10x. Bottom row right: TRPV3 immunostaining in S1BF (inset from top row left) at 10x.

Article Snippet: The following mouse strains were obtained from The Jackson Laboratory (JAX): C57BL/6J (JAX 000664), FVB/N mice ( JAX 004828 ), and Trpv3 (JAX 010773) (Moussaieff A et al., 2008; ).

Techniques: Immunostaining, Incubation, Blocking Assay, Staining

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording whole-cell TRPV3 currents at 30°C (black), 36°C (gray), and 39°C (red) in cortical excitatory pyramidal neurons (PNs) with bath application of camphor (5 mM), a TPRV3 agonist. ( B ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black), 36°C (gray) and 39°C (red) in wild-type (WT) mice: 11 cells from four mice. ( C ) Scatter dot plots of the current density-voltage measurements. ( D ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black) in the presence of camphor (5 mM), a TPRV3 agonist, or camphor (5 mM) + TRPV3 blocker (Forsythoside B, 50 µM) (blue). ( E ) Same as ( D ) but for 36°C. ( F ) Same as ( D ) but for 39°C. ( G ) Current density-voltage (I–V) plot showing the net TRPV3 current (opener – (opener+blocker) condition). In B - F , statistical significance was assessed using a two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: The following mouse strains were obtained from The Jackson Laboratory (JAX): C57BL/6J (JAX 000664), FVB/N mice ( JAX 004828 ), and Trpv3 (JAX 010773) (Moussaieff A et al., 2008; ).

Techniques:

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potential and spiking in an excitatory cortical pyramidal neuron (PN) with an intracellular blocker of TRPV3 channels (Forsythoside B, 50 µM) (left) or TRPV4 channels (RN1734, 10 µM) (right) at just-subthreshold V m at 30°C, 36°C, and 39°C in mouse S1 cortex. ( B ) Percentages of cell types obtained from experiment in A . ( C ) Evoked spikes in L2/3 cortical PNs during temperature elevations to 30°C, 36°C, and 39°C under three conditions: no blockers, TRPV3 blocker (Forsythoside B, 50 µM), or TRPV4 blocker (RN1734, 10 µM). ( D ) Correlation between post-synaptic potential (PSP) peak and spike threshold (ST). r =Pearson correlation coefficient with Deming linear regression. ( E ) Same as ( C ) for the L4-evoked late PSP peak. ( F ) Same as ( C ) for input resistance (R in ). Each data point in C – F represents an individual cell. Data were collected from 26 cells in 7 animals for the TRPV3 blocker, 24 cells in 6 animals for the TRPV4 blocker, and 37 cells in 14 animals for the no-block condition. Mean ± SEM is shown in C , E , and F . Statistical significance was assessed using one- or two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). In D , correlations were evaluated using Pearson’s r with Deming linear regression.

Article Snippet: The following mouse strains were obtained from The Jackson Laboratory (JAX): C57BL/6J (JAX 000664), FVB/N mice ( JAX 004828 ), and Trpv3 (JAX 010773) (Moussaieff A et al., 2008; ).

Techniques: Blocking Assay

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potentials and spiking in excitatory cortical pyramidal neurons (PNs) at just-subthreshold Vm at 30°C, 36°C, and 39°C in mouse S1 cortex of wild-type ( Trpv3 +/+ ) and Trpv3 knockout ( Trpv3 -/- ) mice. ( B ) Depolarization required to reach spike threshold (ST) in Trpv3 +/+ and Trpv3 -/- mice at 30°C, 36°C, and 39°C. ( C ) Same as ( B ) but for input resistance (R in ). ( D ) Same as ( B ) but for number of spikes. ( E ) Same as ( B ) but for post-synaptic potential (PSP). ( F ) Setup for recording mouse body temperature (T b ) at room temperature and during fever-range and higher, using an implanted transponder for non-invasive measurement, with exposure to infrared light. ( G ) Time to loss of postural control (LPC), defined as collapse and failure to maintain upright posture, in wild-type ( Trpv3 +/+ ), heterozygous ( Trpv3 +/- ), and Trpv3 knockout ( Trpv3 -/- ) mice. ( H ) Same as in ( G ) but showing T b at seizure onset. ( I ) Same as in ( G ) but for the time from LPC to seizure onset. Each data point in B – E represents an individual cell (three animals per genotype). Statistical significance was assessed using two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). Each data point in G – I represents an individual animal. Statistical significance was assessed using one-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: The following mouse strains were obtained from The Jackson Laboratory (JAX): C57BL/6J (JAX 000664), FVB/N mice ( JAX 004828 ), and Trpv3 (JAX 010773) (Moussaieff A et al., 2008; ).

Techniques: Knock-Out, Control

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) In cortical L2/3 pyramidal neurons (PNs) with synaptically evoked spiking, gradual increases in brain slice temperature from 30 °C to 36°C to 39°C result in four possible outcomes: neurons remain inactive, continue spiking (STAY), stop spiking, or initiate spiking. To spike, PNs must reach the spike threshold (ST), defined as the minimal V m that elicits an action potential, which requires sufficient depolarization via the post-synaptic potential (PSP). STAY neurons consistently reach ST and continue spiking. Neurons that stop spiking fall below the level of depolarization required to reach ST, while neurons that initiate spiking achieve sufficient depolarization to newly reach ST. ( B ) STAY neurons may contain unique ion channels, such as TRPV3. TRPV3 channels are highly permeable to Ca² + ions, and Ca² + influx contributes to PN depolarization. The presence of TRPV3 facilitates greater ion entry, enabling depolarization sufficient to reach ST and sustain spiking, whereas its absence reduces depolarization to levels insufficient for spiking.

Article Snippet: The following mouse strains were obtained from The Jackson Laboratory (JAX): C57BL/6J (JAX 000664), FVB/N mice ( JAX 004828 ), and Trpv3 (JAX 010773) (Moussaieff A et al., 2008; ).

Techniques: Slice Preparation

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) TRPV3 immunostaining (green) in mouse primary somatosensory (S1) cortex, striatum, thalamus, and hippocampus, with DAPI counterstain (blue). Top row left: Postnatal day (P)7; bottom row left: P14; bottom row right: P21. Top row right: Sections incubated with TRPV3 antibody plus TRPV3 blocking peptide in S1 cortex, striatum, thalamus, and hippocampus (green) with DAPI counterstain (blue). Remaining TRPV3 staining is shown in green with DAPI in blue. ( B ) TRPV3 and TRPV4 immunostaining in S1 cortex, striatum, thalamus, and hippocampus at P14. Top row left: TRPV3 (green) and TRPV4 (brick red). Top row right: TRPV3 (green) and TRPV4 (brick red) with DAPI counterstain (blue). Bottom row left: TRPV4 immunostaining in S1BF (inset from top row left) at 10x. Bottom row right: TRPV3 immunostaining in S1BF (inset from top row left) at 10x.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Immunostaining, Incubation, Blocking Assay, Staining

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording whole-cell TRPV3 currents at 30°C (black), 36°C (gray), and 39°C (red) in cortical excitatory pyramidal neurons (PNs) with bath application of camphor (5 mM), a TPRV3 agonist. ( B ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black), 36°C (gray) and 39°C (red) in wild-type (WT) mice: 11 cells from four mice. ( C ) Scatter dot plots of the current density-voltage measurements. ( D ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black) in the presence of camphor (5 mM), a TPRV3 agonist, or camphor (5 mM) + TRPV3 blocker (Forsythoside B, 50 µM) (blue). ( E ) Same as ( D ) but for 36°C. ( F ) Same as ( D ) but for 39°C. ( G ) Current density-voltage (I–V) plot showing the net TRPV3 current (opener – (opener+blocker) condition). In B - F , statistical significance was assessed using a two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques:

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potential and spiking in an excitatory cortical pyramidal neuron (PN) with an intracellular blocker of TRPV3 channels (Forsythoside B, 50 µM) (left) or TRPV4 channels (RN1734, 10 µM) (right) at just-subthreshold V m at 30°C, 36°C, and 39°C in mouse S1 cortex. ( B ) Percentages of cell types obtained from experiment in A . ( C ) Evoked spikes in L2/3 cortical PNs during temperature elevations to 30°C, 36°C, and 39°C under three conditions: no blockers, TRPV3 blocker (Forsythoside B, 50 µM), or TRPV4 blocker (RN1734, 10 µM). ( D ) Correlation between post-synaptic potential (PSP) peak and spike threshold (ST). r =Pearson correlation coefficient with Deming linear regression. ( E ) Same as ( C ) for the L4-evoked late PSP peak. ( F ) Same as ( C ) for input resistance (R in ). Each data point in C – F represents an individual cell. Data were collected from 26 cells in 7 animals for the TRPV3 blocker, 24 cells in 6 animals for the TRPV4 blocker, and 37 cells in 14 animals for the no-block condition. Mean ± SEM is shown in C , E , and F . Statistical significance was assessed using one- or two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). In D , correlations were evaluated using Pearson’s r with Deming linear regression.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Blocking Assay

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potentials and spiking in excitatory cortical pyramidal neurons (PNs) at just-subthreshold Vm at 30°C, 36°C, and 39°C in mouse S1 cortex of wild-type ( Trpv3 +/+ ) and Trpv3 knockout ( Trpv3 -/- ) mice. ( B ) Depolarization required to reach spike threshold (ST) in Trpv3 +/+ and Trpv3 -/- mice at 30°C, 36°C, and 39°C. ( C ) Same as ( B ) but for input resistance (R in ). ( D ) Same as ( B ) but for number of spikes. ( E ) Same as ( B ) but for post-synaptic potential (PSP). ( F ) Setup for recording mouse body temperature (T b ) at room temperature and during fever-range and higher, using an implanted transponder for non-invasive measurement, with exposure to infrared light. ( G ) Time to loss of postural control (LPC), defined as collapse and failure to maintain upright posture, in wild-type ( Trpv3 +/+ ), heterozygous ( Trpv3 +/- ), and Trpv3 knockout ( Trpv3 -/- ) mice. ( H ) Same as in ( G ) but showing T b at seizure onset. ( I ) Same as in ( G ) but for the time from LPC to seizure onset. Each data point in B – E represents an individual cell (three animals per genotype). Statistical significance was assessed using two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). Each data point in G – I represents an individual animal. Statistical significance was assessed using one-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Knock-Out, Control

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) In cortical L2/3 pyramidal neurons (PNs) with synaptically evoked spiking, gradual increases in brain slice temperature from 30 °C to 36°C to 39°C result in four possible outcomes: neurons remain inactive, continue spiking (STAY), stop spiking, or initiate spiking. To spike, PNs must reach the spike threshold (ST), defined as the minimal V m that elicits an action potential, which requires sufficient depolarization via the post-synaptic potential (PSP). STAY neurons consistently reach ST and continue spiking. Neurons that stop spiking fall below the level of depolarization required to reach ST, while neurons that initiate spiking achieve sufficient depolarization to newly reach ST. ( B ) STAY neurons may contain unique ion channels, such as TRPV3. TRPV3 channels are highly permeable to Ca² + ions, and Ca² + influx contributes to PN depolarization. The presence of TRPV3 facilitates greater ion entry, enabling depolarization sufficient to reach ST and sustain spiking, whereas its absence reduces depolarization to levels insufficient for spiking.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Slice Preparation

Journal: Molecular Pharmacology

Article Title: Activation of TRPV3 channels in bladder cancer cells stimulates ATP release

doi: 10.1016/j.molpha.2025.100096

Figure Lengend Snippet: AV3-1 activated human and mouse TRPV3. (A, B) Concentration-response curves depicting TRPV3 activation by AV3-1 in HEK cells expressing either mouse TRPV3 (A, black squares) or human TRPV3 (B, black squares). Parental HEK cells lacking TRPV3 expression (A, black triangles) served as a negative control and showed no response to AV3-1. Fluorescence signals (F) were normalized to the fluorescence before compound addition (F 0 ). Data are presented as mean ± SD from 5 independent experiments, with duplicates each. Chemical structure of AV3-1 is shown as in inset in (A). (C–E) Microfluorometric single-cell analysis of [Ca 2+ ] i in fura-2-loaded cells. Shown are representative traces of [Ca 2+ ]ᵢ over time in parental HEK cells in response to application of 50 μ M AV3-1 (C) and in HEK cells stably expressing mouse TRPV3 (HEK mTRPV3 ), in response to application of 5 μ M AV3-1, followed by addition of the inhibitor 26E01 (50 μ M) (left panel) or without inhibitor (right panel) (D, E). Individual cell traces are shown in gray; the average trace is overlaid in black. Application periods are indicated by colored bars. (E) Quantification of peak [Ca 2+ ]ᵢ responses from HEK mTRPV3 cells under the indicated treatment conditions. A baseline was recorded in all experiments, followed by AV3-1 application; in 50% of experiments, 26E01 was subsequently added to the cells. Because of this design, sample sizes differ across conditions; the exact n for each box is indicated in the figure. Quantification for parental HEK cells is shown as an inset in (C) (n = 5 independent experiments). Statistical significance was determined using one-way ANOVA with Tukey post hoc analysis; ∗ P < .05. ns, not significant.

Article Snippet: Human TRPV3 ( NM_145068 , purchased from OriGene) and mouse TRPV3 ( NM_001371006 ), rat TRPV1, mouse TRPV2, mouse TRPV4, human TRPM2, human TRPM8, and human TRPA1 were stably transfected in HEK293 cells as described before.

Techniques: Concentration Assay, Activation Assay, Expressing, Negative Control, Fluorescence, Single-cell Analysis, Stable Transfection

Journal: Molecular Pharmacology

Article Title: Activation of TRPV3 channels in bladder cancer cells stimulates ATP release

doi: 10.1016/j.molpha.2025.100096

Figure Lengend Snippet: Electrophysiological analysis of AV3-1–activated TRPV3 currents in HEK m TRPV3 cells. (A) Representative time course of a whole-cell patch-clamp recording obtained from a HEK m TRPV3 cell, showing current responses to repeated applications of the TRPV3 agonist AV3-1 (5 μ M, blue bars, 1 minute each), followed by a washout period. The TRPV3 inhibitor 26E01 (50 μ M, red bar) was applied during the third AV3-1 stimulation. Current amplitudes were recorded using voltage ramps from −100 mV to +100 mV (500-millisecond duration) such as in B and expressed as current density (pA/pF) measured at +100 mV and −100 mV. The labels (1, 6–8) indicate time points for statistical analysis as shown in (C). (B) Representative current-voltage (I–V) curves from the same cell shown in (A), taken at baseline (1, black), at peak response to AV3-1 prior to 26E01 addition (6, blue), and during coapplication with 26E01 (7, red). (C) Quantification of current densities (pA/pF) at −100 mV (inward current) and +100 mV (outward current) at the time points (1, 6–8) as indicated in (A). Box plot shows data of n = 5 independent recordings. Statistical significance was determined using Kruskal-Wallis ANOVA followed by Conover test; ∗ P < .05. Asterisks indicate significance for both outward and inward current components.

Article Snippet: Human TRPV3 ( NM_145068 , purchased from OriGene) and mouse TRPV3 ( NM_001371006 ), rat TRPV1, mouse TRPV2, mouse TRPV4, human TRPM2, human TRPM8, and human TRPA1 were stably transfected in HEK293 cells as described before.

Techniques: Patch Clamp

Journal: Molecular Pharmacology

Article Title: Activation of TRPV3 channels in bladder cancer cells stimulates ATP release

doi: 10.1016/j.molpha.2025.100096

Figure Lengend Snippet: TRPV3 is functionally expressed in bladder cancer cells. (A–D) Activation of TRPV3 in KU-19-19 (A, B) and CAL-29 (C, D) bladder cancer cells by AV3-1, assessed by fura-2-based Ca 2+ assays. The experimental layout was as in , D and . Shown are representative traces of [Ca 2+ ]ᵢ over time in response to application of 50 μ M AV3-1, followed by addition of the inhibitor 26E01 (50 μ M) (left panel) or without inhibitor (right panel). Individual cell traces are shown in gray; the average trace is overlaid in black. Application periods are indicated by colored bars. (B, D) Quantification of peak [Ca 2+ ]ᵢ responses from KU-19-19 (B) and CAL-29 (D) cells under the indicated treatment conditions. The exact n for each box is indicated in the figure. Statistical significance was determined using one-way ANOVA with Tukey post hoc analysis; ∗ P < .05. (E–G) Electrophysiological whole-cell recordings performed on KU-19-19 cells. (E) Representative time course of a whole-cell patch-clamp recording obtained from a KU-19-19 cell, showing current responses to repeated applications of the TRPV3 agonist AV3-1 (50 μ M, blue bars, 1 minute each). The TRPV3 inhibitor 26E01 (50 μ M, red bar) was applied during the third AV3-1 stimulation. Current amplitudes were recorded using voltage ramps from −100 mV to +100 mV (500-millisecond duration) such as in F and expressed as current density (pA/pF) measured at +100 mV and −100 mV. The labels (1, 6–8) indicate time points for statistical analysis as shown in (G). (F) Representative current-voltage (I–V) curves from the same cell shown in (E), taken at baseline (1, black), at peak response to AV3-1 prior to 26E01 addition (6, blue), and during c-application with 26E01 (7, red). (G) Quantification of current densities (pA/pF) at −100 mV (inward current) and +100 mV (outward current) at the time points (1, 6–8) as indicated in (E). Box plot shows data of n = 5 independent recordings. Statistical significance was determined using Kruskal-Wallis ANOVA followed by Conover test; ∗ P < .05. Asterisks indicate significance for both outward and inward current components.

Article Snippet: Human TRPV3 ( NM_145068 , purchased from OriGene) and mouse TRPV3 ( NM_001371006 ), rat TRPV1, mouse TRPV2, mouse TRPV4, human TRPM2, human TRPM8, and human TRPA1 were stably transfected in HEK293 cells as described before.

Techniques: Activation Assay, Patch Clamp

Journal: Molecular Pharmacology

Article Title: Activation of TRPV3 channels in bladder cancer cells stimulates ATP release

doi: 10.1016/j.molpha.2025.100096

Figure Lengend Snippet: TRPV3 activation by AV3-1 was enhanced by cholesterol supplementation and causes ATP release in KU-19-19 bladder cancer cells. (A–C) Electrophysiological whole-cell recordings performed on KU-19-19 cells without (A) and with (B) cholesterol supplementation. (A, B) Representative time courses of whole-cell patch-clamp recordings obtained from a KU-19-19 cell, showing current responses to repeated applications of the TRPV3 agonist AV3-1 (50 μ M, blue bars, 1 minute each). Current amplitudes were recorded using voltage ramps from −100 mV to +100 mV (500-millisecond duration) and expressed as current density (pA/pF) measured at +100 mV and −100 mV. (C) Quantification of current densities (pA/pF) at −100 mV (inward current) and +100 mV (outward current) at the time points (1, 2) as indicated in (A, B). Box plot show data of n = 10 independent recordings. Statistical significance was determined using Kruskal-Wallis ANOVA followed by Conover test; ∗ P < .05. Asterisks indicate significance for both outward and inward current components. (D) Concentration-response curves depicting TRPV3 activation in KU-19-19 cells without (open squares) or with (gray circles) cholesterol supplementation. Experiments were conducted as described in , A and , and depict means ± SD of 5 independent experiments. Inset: Box-and-whisker plots of AV3-1 potencies under control conditions (open box) and following cholesterol supplementation (gray box), derived from individual concentration-response fits and expressed as pEC 50 (-log 10 (EC 50 [M])). Each diamond represents an individual experiment (n = 5 per condition). Group differences were tested on pEC 50 values using a two-tailed unpaired Student t test; ∗ P < .05. (E) Concentration response analysis of AV3-1 in MTT assays with KU-19-19 cells depicting cell viability and proliferation after 24 hours of stimulation with AV3-1 at concentrations as indicated. Data represent means ± SD of n = 5 independent experiments. (F) Box plot quantification of extracellular ATP levels in KU-19-19 cells after stimulation with AV3-1 (50 μM, blue), measured by a luciferin-luciferase–based luminescence assay. Luminescence values are normalized to control values observed in wells containing only media (L/L control). Pretreatment with the TRPV3 antagonist 26E01 (50 μ M, red) or the nonselective TRP channel blocker ruthenium red (RR; 10 μM, red diagonal hatched fill) reduced AV3-1–evoked ATP release. Statistical analysis of n = 6 independent measurements was performed using one-way ANOVA with Tukey post hoc test; ∗ P < .05. ns, not significant.

Article Snippet: Human TRPV3 ( NM_145068 , purchased from OriGene) and mouse TRPV3 ( NM_001371006 ), rat TRPV1, mouse TRPV2, mouse TRPV4, human TRPM2, human TRPM8, and human TRPA1 were stably transfected in HEK293 cells as described before.

Techniques: Activation Assay, Patch Clamp, Concentration Assay, Whisker Assay, Control, Derivative Assay, Two Tailed Test, Luciferase, Luminescence Assay

Journal: Molecular Pharmacology

Article Title: Transient receptor potential vanilloid 3 activation accelerates keratinocyte migration in vitro but not dermal wound healing in vivo

doi: 10.1016/j.molpha.2025.100084

Figure Lengend Snippet: WT keratinocytes close gaps statistically significantly faster than TRPV3 KO keratinocytes in vitro. (A, B) Primary keratinocytes from WT (A) and TRPV3 KO (B) mice closing a 500- μ m gap (start at t = 0 hours) after 18, 24, and 30 hours of migration and proliferation. (C) Statistical analysis of the remaining gap areas as shown in (A) and (B) of WT (blue boxes) and TRPV3 KO (red boxes) keratinocytes after 18, 24, and 30 hours. Cells were kept in keratinocyte growth medium at 37 °C and 5% CO 2 in a humidified atmosphere (n WT = 12, n TRPV3 KO = 13, ∗ P ≤ .05, data not normally distributed).

Article Snippet: Adult C57BL/6J WT (The Jackson Laboratory, RRID: MSR_JAX:000664) and genetically matching TRPV3 KO (B6.129- Trpv3 tm1Apat /J, RRID:IMSR_JAX:010773) mice were housed in groups of 2–3 animals under approved standard conditions of a 12-hour light–dark regime and access to food and water ad libitum.

Techniques: In Vitro, Migration

Journal: Molecular Pharmacology

Article Title: Transient receptor potential vanilloid 3 activation accelerates keratinocyte migration in vitro but not dermal wound healing in vivo

doi: 10.1016/j.molpha.2025.100084

Figure Lengend Snippet: AV3-1-induces Ca 2+ sparklets in WT keratinocytes. (A, D) WT (A) and TRPV3 KO (D) keratinocytes were loaded with the low-affinity Ca 2+ indicator Cal520ff, and imaged by gap-free TIRF microscopy at 20 frames per second. (B, C, E, F) Maximum projection of the variance analysis of the corresponding raw data shown in (A) and (D) within 50 seconds before AV3-1 addition (B, E) and within 50 seconds after AV3-1 (3 μ M) addition (C, F). Note the preferred localization of Ca 2+ sparklets in the leading edges of WT keratinocytes (C). The intensity variance was calculated over consecutive bins of 10 frames and divided by the respective mean fluorescence to obtain a measure of local Ca 2+ fluctuations as represented by a pseudocolor scale. The maxima of 100 bins before and after AV3-1 addition, respectively, are projected in each image. (G) Quantitative analysis of the appearing sparklets per second and mm 2 cell area in WT (blue boxes) and TRPV3 KO (red boxes) keratinocytes (n WT = 9, n TRPV3 KO = 14, ∗ P ≤ .05, data not normally distributed). n.s., statistically not significant.

Article Snippet: Adult C57BL/6J WT (The Jackson Laboratory, RRID: MSR_JAX:000664) and genetically matching TRPV3 KO (B6.129- Trpv3 tm1Apat /J, RRID:IMSR_JAX:010773) mice were housed in groups of 2–3 animals under approved standard conditions of a 12-hour light–dark regime and access to food and water ad libitum.

Techniques: Microscopy, Fluorescence

Journal: Molecular Pharmacology

Article Title: Transient receptor potential vanilloid 3 activation accelerates keratinocyte migration in vitro but not dermal wound healing in vivo

doi: 10.1016/j.molpha.2025.100084

Figure Lengend Snippet: TRPV3 activation stimulates PI3 kinases in an EGFR-dependent manner. (A, B) TIRF microscopy signals of WT (A) and TRPV3 KO (B) keratinocytes transfected with 4 μ L LipofectAMINE 2000 and 1 μ g cDNA encoding YFP-GRP1(PH), a translocating biosensor of phosphatidylinositol (3,4,5)-trisphosphate formation. Blue bars indicate AV3-1 addition (15 μ M). (C) Statistical analysis of the TIRFM signals evoked by AV3-1, as shown in (A) and (B) in WT (blue boxes) and TRPV3 KO (red boxes) keratinocytes. EGFR in WT cells was inhibited by AG 1478 (100 nM) in a second set of experiments (n WT = 9, n KO = 12, n AG 1478 = 24, data not normally distributed). (D) WT keratinocytes were transfected with 4 μ L LipofectAMINE 3000 and 1 μ g cDNA encoding YFP-GRP1(PH) in a third set of experiments. The graph analyses the statistical fluorescence increase caused by the addition of AV3-1 (15 μ M) without and with prior inhibition of EGFR by erlotinib (10 μ M, n control = 23, n erlotinib = 36, ∗ P ≤ .05, data not normally distributed).

Article Snippet: Adult C57BL/6J WT (The Jackson Laboratory, RRID: MSR_JAX:000664) and genetically matching TRPV3 KO (B6.129- Trpv3 tm1Apat /J, RRID:IMSR_JAX:010773) mice were housed in groups of 2–3 animals under approved standard conditions of a 12-hour light–dark regime and access to food and water ad libitum.

Techniques: Activation Assay, Microscopy, Transfection, Fluorescence, Inhibition, Control

Journal: Molecular Pharmacology

Article Title: Transient receptor potential vanilloid 3 activation accelerates keratinocyte migration in vitro but not dermal wound healing in vivo

doi: 10.1016/j.molpha.2025.100084

Figure Lengend Snippet: WT and TRPV3 KO mice display no statistically significant difference in dermal wound areas over 5 days in vivo. Remaining wound areas in WT (blue boxes) and TRPV3 KO (red boxes) mice, at the indicated number of days after biopsy. On day 0, two 5-mm biopsy punch wounds were excised on the dorsal skin of the mice, equally distanced on each side of the midline (n WT = 6, n TRPV3 KO = 6, data not normally distributed).

Article Snippet: Adult C57BL/6J WT (The Jackson Laboratory, RRID: MSR_JAX:000664) and genetically matching TRPV3 KO (B6.129- Trpv3 tm1Apat /J, RRID:IMSR_JAX:010773) mice were housed in groups of 2–3 animals under approved standard conditions of a 12-hour light–dark regime and access to food and water ad libitum.

Techniques: In Vivo

Journal: Molecular Pharmacology

Article Title: Transient receptor potential vanilloid 3 activation accelerates keratinocyte migration in vitro but not dermal wound healing in vivo

doi: 10.1016/j.molpha.2025.100084

Figure Lengend Snippet: Pharmacological activation of TRPV3 does not induce a statistically significant acceleration of dermal wound healing with or without topical application of EGF. Remaining wound areas in WT (blue boxes) and TRPV3 KO (red boxes) animals over 5 days are shown. Mice were either treated with AV3-1 (100 μ M) or the combination of AV3-1 (100 μ M) and EGF (10 ng/mL). The treatments were applied on one of the 5-mm biopsy punch wounds, whereas the contralateral wound received the corresponding vehicle or EGF alone (10 ng/mL) ( n = 5 for each group, ∗ P ≤ .05, data not normally distributed).

Article Snippet: Adult C57BL/6J WT (The Jackson Laboratory, RRID: MSR_JAX:000664) and genetically matching TRPV3 KO (B6.129- Trpv3 tm1Apat /J, RRID:IMSR_JAX:010773) mice were housed in groups of 2–3 animals under approved standard conditions of a 12-hour light–dark regime and access to food and water ad libitum.

Techniques: Activation Assay

Journal: Molecular Pharmacology

Article Title: Transient receptor potential vanilloid 3 activation accelerates keratinocyte migration in vitro but not dermal wound healing in vivo

doi: 10.1016/j.molpha.2025.100084

Figure Lengend Snippet: AV3-1 treatment with or without co-application of EGF causes no statistically significant increase in the extent of reepithelialization. (A) RGB-stained slice of a wound on day 5 postsurgery with the following analyzed wound parameters: wound area (orange dotted line), epidermal area (green dotted line), and wound length and epidermal length (indicated by arrows). The insert shows the boundary between the scab and epidermis. (B) Statistical analysis of the epidermal area ratio (epidermal area/wound area) and the epidermal length ratio (epidermal length/wound length) of WT (blue boxes) and TRPV3 KO (red boxes) wounds as shown exemplary in (A) (n WT, DMSO = 3, n WT, AV3-1 = 3, n WT, EGF = 5, n WT, AV3-1, EGF = 5, n TRPV3 KO, EGF = 5, n TRPV3 KO, AV3-1, EGF = 5, data normally distributed).

Article Snippet: Adult C57BL/6J WT (The Jackson Laboratory, RRID: MSR_JAX:000664) and genetically matching TRPV3 KO (B6.129- Trpv3 tm1Apat /J, RRID:IMSR_JAX:010773) mice were housed in groups of 2–3 animals under approved standard conditions of a 12-hour light–dark regime and access to food and water ad libitum.

Techniques: Staining