Combined Changes in Chloride Regulation and Neuronal Excitability Enable Primary Afferent Depolarization to Elicit Spiking without Compromising its Inhibitory Effects
Fig 6
ANO-1 channels do not contribute to PAD.
For all panels, responses in the presence of the ANO-1 antagonist T16Ainh-A01 (A01) are shown in red for comparison against responses in normal aCSF shown in black. (A) Traces show responses in a typical neuron to the minimum virtual ḡGABA required to evoke spiking based on a fast synaptic waveform and EGABA = -20 mV before and after ANO-1 blockade. Summary data show that the minimum ḡGABA to evoke spiking was not significantly changed by A01 (p = 1.0; paired t-test) based on all TRPV1+ neurons (n = 5) that spiked in response to virtual GABA. (B) Traces show responses in a typical neuron to different ḡGABA based on slow synaptic waveform and EGABA = -35 mV. Summary data show mean (± SEM) depolarization at different ḡGABA for all (n = 7) TRPV1+ neurons tested. Blockade of ANO-1 did not significantly affect depolarization (p = 0.77; two-way repeated measure ANOVA). (C) At the end of each experiment, the recorded cell was stimulated with capsaicin. Traces show typical data from a responsive (TRPV1+) and unresponsive (TRPV1-) neuron. Because ANO-1 is expressed predominantly in TRPV1+ neurons, only data from capsaicin-responsive neurons were included for analysis in panels A and B. (D) To confirm the efficacy A01, we verified that it reduced the response to capsaicin, consistent with Takayama et al. [49].