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. 2025 Jan 10;16:584. doi: 10.1038/s41467-025-55936-5

Fig. 2. Whole-cell patch-clamp recordings of mouse tracheal tuft cells.

Fig. 2

A Application of denatonium (1 mM) in Trpm5+/+ tuft cells led to an outwardly rectifying current that was abolished by TPPO (100 µM). B Denatonium (1 mM) had no effect on the I/V curve recorded in Trpm5−/− tuft cells. Red = denatonium, black = baseline. C Current densities recorded at 100 mV were increased by denatonium in tuft cells of Trpm5+/+ mice compared to before treatment and to currents in the presence of 100 µM TPPO (n = 4 cells/3 mice, paired Student’s t-test). Currents at 100 mV of Trpm5−/− tuft cells (n = 11 cells/5 mice) perfused with 1 mM denatonium were reduced compared to tuft cells from Trpm5+/+ mice (two-tailed unpaired Student’s t-test). D Representative current traces at a holding potential of +30 mV hyper-polarized to −60 mV upon application of 60 mM KCl and addition of 300 µM probenecid (prob). E KCl-induced current densities were inhibited by carbenoxolone (CBX, 10 µM) or probenecid (prob, 300 µM) in tuft cells from Trpm5+/+ and Trpm5−/− mice. Trpm5+/+ CBX: n = 3 cells/3mice, prob: n = 9 cells/5mice, Trpm5−/− CBX: n = 8 cells/3mice, prob: n = 8 cells/3mice, two-tailed paired Student’s t-test. F The KCl-induced current densities in Trpm5+/+ (n = 13 cells/7mice) and Trpm5−/− (n = 16 cells/5mice) mice were identical (two-tailed unpaired Student’s t-test). G Analyzed current densities at 100 mV upon a ramp depolarization. Carbenoxolone (CBX, 10 µM)-sensitive currents upon the activation of Trpm5 channels with 110 nM [Ca2+]i were reduced in Trpm5+/+ but not in Trpm5−/− mice. Trpm5 currents at 110 nM [Ca2+]i and 2.4 µM [Ca2+]i were sensitive to probenecid (prob, 300 µM) in Trpm5+/+ but not in Trpm5−/− mice. CBX: Trpm5+/+: n = 9 cells/6mice, Trpm5−/−: n = 10 cells/5mice, prob, 110 nM Ca2+: Trpm5+/+: n = 12 cells/4mice, Trpm5−/−: n = 7 cells/3mice, prob, 2.4 µM Ca2+: Trpm5+/+: n = 13 cells/4mice, Trpm5−/−: n = 8 cells/3mice; one-tailed paired Student’s t-test. H The Ca2+-induced current densities in tuft cells (ctrl) were not inhibited by the P2x7 receptor inhibitor AZ10606121 (20 µM) (n = 8 cells/4 mice/4 experiments). Two-tailed Wilcoxon test. Source data are provided in the Source Data file. Data in panels F and H are presented as mean ± SEM.