Role of CGRP in KI P2X3 currents.

<p>A, Representative traces of currents induced by application of α,β-meATP (10 µM, 2 s) to WT or R192Q KI neurons in control conditions or after 2 h CGRP (1 µM) application. B, Histograms show average peak amplitudes of P2X3 receptor-mediated currents (WT control, n = 11; WT CGRP, n = 9; KI control, n = 17; KI CGRP, n = 18), * = p<0.05. C, Representative examples of currents induced by application of α,β-meATP (10 µM, 2 s) to WT or R192Q KI neurons in control conditions or after overnight application of the CGRP antagonist CGRP_8–37 (1 µM). D, Histograms show average peak amplitudes of P2X3 receptor-mediated currents (WT control, n = 23; WT CGRP_8–37, n = 23; KI control, n = 29; KI CGRP_8–37, n = 30); * = p<0.05.</p

Number of BDNF expressing neurons in KI culture and effect of BDNF deprivation.

<p>A, Examples of β-tubulin positive neurons expressing BDNF in WT or KI cultures. Left panel (green) shows BDNF expression and right panel (red) shows β-tubulin staining of the same neurons. B, Histograms quantifying % of neurons expressing BDNF: KI cultures showed significantly higher number of BDNF positive neurons. N = 4 independent experiments (8 mice), p<0.05. C, Representative traces of currents induced by application of α,β-meATP (10 µM, 2 s) to WT or R192Q KI neurons in control conditions or after overnight application of anti-BDNF antibody. D, Histograms show average peak amplitudes of P2X3 receptor-mediated currents (WT control, n = 9; WT anti-BDNF, n = 10; KI control, n = 32; KI anti-BDNF, n = 38); * = p<0.05.</p

Sensitization of TRPV1 receptors by TNFα.

<p>A, Representative traces of currents induced by application of capsaicin (1 µM, 2 s) to WT or R192Q KI neurons in control conditions or after 4 h application of TNFα. B, Histograms show average peak amplitudes of TRPV1-mediated currents (WT control, n = 29; WT TNFα 50 ng/mL n = 19, WT TNFα 100 ng/mL, n = 10; KI control, n = 26; KI TNFα 50 ng/mL, n = 23,, KI TNFα 100 ng/mL n = 9); * = p<0.05, ** = p<0.002, *** = p<0.001.</p

Effect of TNFα on P2X3 receptor activity and co-expression of its TNFR2 receptors.

<p>A, Examples of TNFR2 and P2X3 co-exexpression in (wildtype) WT and R192Q (knockin) KI neurons. Left panel shows P2X3 expression (green), and right panel shows TNFR2 staining (red). B, Histograms quantifying % of cells co-expressing TNFR2 and P2X3: both WT and KI cultures show similar TNFR2 and P2X3 co-expression. N = 3 independent experiments (6 mice). C, Representative traces of currents induced by application of α,β-meATP (10 µM, 2 s) to WT or R192Q KI neurons in control conditions or after 4 h TNFα application. D, Histograms show average peak amplitudes of P2X3 receptor-mediated currents: WT control (open bar), n = 30; WT TNFα (stippled bar), n = 38; KI control (grey bar), n = 34; KI TNFα (stippled gray bar), n = 34; ** = p<0.006; * = p<0.05.</p