Article Figures & Data
Figures
- Figure 1 The TRPM8-R30Q Enhances Calcium Entry in Response to Stimulation by Menthol
Average changes in the Fura-2 ratio of human embryonic kidney 293 cells transfected with wild-type TRPM8 (A) and R30Q mutant (B), in the continued presence of menthol (100 μM; arrow indicates time of addition). (C) Average increase in Fura-2 ratio in response to menthol (100 μM) in nontransfected cells (NT), wild-type TRPM8, and R30Q transfected cells (n = 10 experiments, 4 transfections). *p < 0.05, unpaired Student's t test. (D) Basal fura-2 ratio in wild-type TRPM8 and R30Q mutant transfected cells (n = 15 experiments, 4 transfections). *p < 0.05, unpaired Student t test. TRPM8 = transient receptor potential melastatin 8.
- Figure 2 Voltage-Clamp Recordings Show Increased Basal Activity of TRPM8-R30Q Channels
(A) Representative whole-cell current traces through wild-type TRPM8 and R30Q transfected cells, in response to the indicated voltage step protocol. (B) Steady state current-voltage relationships of the basal whole-cell currents for wild-type TRPM8 and R30Q mutant (n = 12, **p < 0.01, ***p < 0.001, 2-way analysis of variance with Bonferroni post hoc test). (C) Steady-state activation curves of wild-type TRPM8 and R30Q transfected cells. The normalized conductance (G/Gmax) was plotted against voltage and fitted with a Boltzmann function, giving rise to V1/2 and slope factor as follows: TRPM8, 153 ± 8 mV and 30 ± 3 mV (n = 12 cells, 3 transfections); R30Q, 115 ± 5 mV and 28 ± 2 mV (n = 12 cells, 3 transfections). **p < 0.01, unpaired Student's t test. (D) Representative time courses (left) recorded at +80 mV and −80 mV (black and grey curves, respectively) and I-V traces (right) of whole cell currents through wild-type TRPM8 transfected cells, in the presence of 100 μM menthol, at the indicated time intervals. (E) Same as D), except that time courses and I-V traces are recorded from R30Q transfected cells. (F) Pooled data of whole-cell current (at +80 mV and −80 mV) evoked by 100 μM menthol, from wild-type TRPM8 and R30Q transfected cells. Each column represents mean ± SEM of n = 8 cells, 3 independent experiments. *p < 0.05 (unpaired Student's t-test). TRPM8 = transient receptor potential melastatin 8.
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