DOI: 10.5061/dryad.t1g1jwtgv
title: Data from: Sex-specific vascular effects of menthol: TRPM8 and TRPA1-dependent relaxation in female mouse aorta and pudendal arteries

description: Sexual dysfunction affects the quality of life for both men and women.
 Menthol, a TRPM8 agonist, is widely used in products to enhance
 sexual performance due to its cooling effect. Beyond this, menthol also
 induces vascular effects in male arteries. To explore its effects on
 female vasculature, we studied male and female C57BL/6 mice, along with
 female TRPM8 knockout (KO) mice. We isolated the internal pudendal artery
 and aorta to assess isometric contractile force. Menthol-induced
 relaxation of the pudendal artery was reduced in both female
 TRPM8 KO and male mice compared to female controls. Acetylcholine-induced
 relaxation was not affected in females, but in males, it was attenuated by
 menthol and enhanced by M8-B, a TRPM8 inhibitor. Menthol did not
 alter responses to norepinephrine, serotonin, or histamine in
 either sex. Notably, menthol-induced relaxation was also reduced
 in the aortas of female TRPM8 KO mice. In both the aorta and
 pudendal arteries of female mice, the TRPA1 inhibitor HC030031
 significantly diminished menthol-induced relaxation.
 These findings suggest that TRPM8 modulation plays a role in
 acetylcholine-induced relaxation in male, but not female,
 pudendal arteries. Moreover, the menthol vascular effect
 in female arteries relies on TRPM8 and TRPA1 activation.
description: # Data from: Sex-specific vascular effects of menthol: TRPM8 and
 TRPA1-dependent relaxation in female mouse aorta and pudendal arteries
 Dataset DOI:
 [10.5061/dryad.t1g1jwtgv](https://doi.org/10.5061/dryad.t1g1jwtgv) ##
 Description of the data and file structure Pudendal and aortic artery
 rings (2 mm) were mounted on wire or pin myographs and maintained in
 oxygenated Krebs–bicarbonate solution at 37 °C; pudendal arteries were
 normalized using dedicated software, whereas aortic rings were set to a
 passive tension of 5 mN. For vascular reactivity studies, arterial rings
 were precontracted with phenylephrine, except during norepinephrine and
 serotonin concentration–response experiments, and cumulative
 concentration–response curves were constructed for norepinephrine,
 serotonin, acetylcholine, histamine, or menthol in the absence or
 following incubation with menthol, M8-B, or HC030031. All data are
 presented as mean ± standard error of the mean (SEM), where *n* denotes
 the number of independent mice. Concentration–response curves with a clear
 sigmoidal profile were analyzed by nonlinear regression, whereas curves
 that did not reach a maximal response or lacked a sigmoidal shape (menthol
 and histamine) are shown as connected data points to illustrate trends
 without extrapolation. Comparisons between experimental groups were
 performed using unpaired Student’s *t* test or two-way analysis of
 variance (ANOVA) followed by Šídák’s multiple-comparisons test, as
 appropriate, with *P* &lt; 0.05 considered statistically significant. For
 two-way ANOVA analyses, differences between concentration–response curves
 were assessed based on the significance of the column (group) factor, as
 indicated by the F value. All statistical analyses were conducted using
 GraphPad Prism software (version 9.5.0; GraphPad Software Inc., La Jolla,
 CA, USA). ### Files and variables #### File: Figure_1.pzf
 **Description:** Percentage of relaxation to cumulative concentrations of
 menthol in pudendal arteries precontracted with phenylephrine
 (1microMolar). A: comparison of female vs male. B: comparison of female
 control vs female TRPM8 KO mice. #### File: Figure_2.pzf
 **Description:** Percentage of relaxation to cumulative concentrations of
 acetylcholine in pudendal arteries precontracted with phenylephrine
 (1microMolar). A: comparison of response in female mice in control vs
 under menthol (30 microMolar) incubation. B: comparison of response in
 male mice in control vs under menthol (30 microMolar) incubation.
 C: comparison of response in female mice in control vs under M8-B (1
 microMolar) incubation. D: comparison of response in male mice in control
 vs under M8-B (1 microMolar) incubation. #### File: Figure_3.pzf
 **Description:** Concentration-response curves to different agonists in
 pudendal arteries in control conditions or incubated with menthol (30
 microMolar). A: Contraction to norepinephrine in females. B: Contraction
 to norepinephrine in males. C: Contraction to serotonin in females. D:
 Contraction to serotonin in males. E: Percentage of relaxation to
 histamine in females preconstricted to phenylephrine (1 microMolar). F:
 Percentage of relaxation to histamine in males preconstricted to
 phenylephrine (1 microMolar). #### File: Figure_4.pzf
 **Description:** Comparison of diameter and vascular reactivity of
 pudendal arteries from female control vs. female TRPM8 KO mice. A:
 Diameter of pudendal arteries. B: Contraction elicited by 80 miliMolar KCl
 solution. C: Concentration-response curve to contraction to
 norepinephrine. D: Concentration-response curve to percentage of
 relaxation acetylcholine in arteries precontracted with phenylephrine (1
 microMolar). #### File: Figure_5.pzf
 **Description:** Concentration-response curve to menthol in segments of
 thoracic aorta precontracted with phenylephrine (10 microMolar). A:
 Comparison of menthol percentage of relaxation in aortas with or without
 endothelium. B: Comparison of menthol percentage of relaxation in aortas
 of female control vs. female TRPM8 KO mice.  #### File: Figure_6.pzf
 **Description:** Concentration-response curve to menthol comparing female
 arteries in the abscence or presence of TRPA1 inhibitor (HC030031, 50
 microMolar). A: Comparison of menthol percentage of relaxation in aortas
 of female control with and without HC030031 precontracted with
 phenylephrine (10 microMolar). B: Comparison of menthol percentage of
 relaxation in aortas of female TRPM8 KO mice with and without HC030031
 precontracted with phenylephrine (10 microMolar). C: Comparison of menthol
 percentage of relaxation in pudendal arteries of female TRPM8 KO mice with
 and without HC030031 precontracted with phenylephrine (1 microMolar). ####
 File: Figure_Supplemental.pzfx **Description:** S1: Comparison of male and
 female pudendal arteries diameter and contractile responsiveness. A:
 Diameter of pudendal arteries. B: Contraction by 80 miliMolar KCl
 solution. C: Concentration-response curve to contraction to
 norepinephrine. D: Concentration-response curve to contraction to
 serotonin. E: Concentration-response curve to percentage of relaxation of
 acetylcholine in pudendal arteries precontracted with phenylephrine (1
 microMolar). F: Concentration-response curve to percentage of relaxation
 to histamine in pudendal arteries precontracted with phenylephrine (1
 microMolar). S2: Comparison of acetylcholine percentage of relaxation in
 aortas with endothelium intact and denuded. ## Code/software N/A ## Access
 information Other publicly accessible locations of the data: * n/a Data
 was derived from the following sources: * n/a