Fig. 4.

Effect of Isok on the Ca²⁺ signals evoked by PregS and pruritogens in Trpm3^+/+ mice. (A) Averaged traces of several PregS+ TG neurons from Trpm3^+/+ mice showing mean changes in intracellular Ca²⁺ concentration in response to 20 μM PregS in the presence or absence of 3 μM IsoK. 25 mM KCl was used as positive control to depolarize the neuronal cell membrane. (B) Statistical analysis of Isok effect on PregS induced Ca²⁺ transients in PregS+ Trpm3^+/+ TG sensory neurons as shown in panel (A). Values are given as percentage of the first PregS-induced Ca²⁺ transient. Dots represent individual neurons, and horizontal lines indicate mean values. Effect of Isok was compared to the vehicle treated control group by Mann-Whitney test, n.s: p > 0.05 (non-significant), ***p < 0.001. (C) Representative traces showing typical changes of intracellular Ca²⁺ concentration in TG neurons from Trpm3^+/+mice in response to 100 μM serotonin and 20 μM PregS in the presence and absence of 3 μM Isok. 25 mM KCl was used as positive control to depolarize the neuronal cell membrane. (D) Statistical analysis of Isok effect on the pruritogens induced Ca²⁺ transients in pruritogen responsive TG sensory neurons from Trpm3^+/+ animals. Values are given as Δ(F1/F0), dots represent individual neurons, and horizontal lines indicate mean values. Effect of Isok was compared to the vehicle treated control group by Mann-Whitney test, n.s: p > 0.05 (non-significant). (E) Percentage of TG neurons form WT mice responding to Hist, 5-HT and ET-1 in the presence and absence of 3 μM Isok. The measurements were carried out as in panel (C). The distribution of the pruritogen responders among TG sensory neurons was compared using Chi squared test, n.s.: p > 0.05 (non-significant). Responders are marked with the indicated colors and non-responders are marked with grey. In each group, neurons were isolated from ≥3 mice and measured in independent experiments.