Fig. 3

TRPA1 antagonist blocks IL-1β induced calcium flux and current in nodose ganglion sensory neurons. a, b Nodose ganglion sensory neurons harvested from VGlut2-cre/GCaMP3 mice were stimulated with IL-1β (20 µg/mL), AITC (100 µM; TRPA1-agonist) and AM0902 (10 µM; TRPA1-antagonist) indicated by the labeled black lines. a Representative calcium traces of sensory neurons. Nodose ganglia neurons fail to respond to IL-1β in the presence of TRPA1-antagonist AM0902. Washing of AM0902 restores sensory neuron responses to both IL-1β and TRPA1-agonist AITC. b Representative calcium traces of sensory neurons responding to IL-1β. Independent of the timing of IL-1β administration, addition of TRPA1-antagonist AM0902 to the culture during blocks IL-1β-induced calcium flux. c, d IL-1β evokes current in TRPA1⁺ sensory neurons. Nodose ganglion sensory neurons harvested from VGlut2-Cre/GCaMP3 mice were first exposed to IL-1β. Florescent imaging was carried out on the microscope to identify IL-1β-responsive neurons in the nodose culture. The cells were washed, and IL-1β-responsive neurons were selectively recorded using patch-clamp and responses to a TRPA1 agonist and antagonist sequentially recorded in individual neurons. c Representative response to IL-1β and TRPA1-agonist AITC. When held at − 70 mV, IL-1β (20 µg/mL) and AITC (100 µM) induced inward currents (arrows), AITC inward currents were blocked by TRPA1 antagonist AM0902 (50 µM). d IL-1β-induced responses were blocked by TRPA1-antagonist. Representative response to IL-1β monitored in a patch-clamp recording of nodose ganglion neurons. When held at − 70 mV, IL-1β (20 µg/mL) induced inward currents (arrow) were blocked by bath application of TRPA1 antagonist AM0902 (50 µM)