Figure 7: Effect of TC-specific knockdown of ANO2 on spike adaptation and mAHP currents in TC neurons.

(a) AHP currents were measured in VB TC neurons infected with Scr or shANO2 AAV viruses. (b) shANO2-infected neurons (n=10 from three mice) displayed significantly lower current densities compared with AAV-Scr-infected neurons (n=6 from six mice; *P=0.014, t-test). (c) Tonic firing and AHP were generated by current injection in TC neurons infected with AAV-Scr or AAV-shANO2. (d) AHP potentials in AAV-Scr- (n=6 from five mice) or AAV-shANO2-infected (n=7 from five mice) TC neurons were quantified (**P=0.009, t-test). (e,f) In shANO2-infected TC neurons, the AHP after current injection decreased, whereas the number of spikes (**P=0.009, t-test) and the adaptation indexes increased (***P=0.0008, t-test). (g) Single action potentials from AAV-Scr (black) and AAV-shANO2 (red) infected TC neurons. (h) There was no difference in the half width of single action potentials between AAV-Scr-infected (black, n=6 from five mice) and AAV-shANO2-infected (red, n=7 from five mice) TC neurons (P=0.46, t-test). (i) Representative traces of burst firing on resting membrane potential (∼−60 mV) released from hyperpolarization (∼−80 mV). (j) Patterns and number of spikes per burst were similar in AAV-Scr-infected (n=6 from five mice) and AAV-shANO2-infected (n=7 from five mice) TC neurons (P=0.57, t-test). (k) Under intact [Cl−]in conditions performed by perforated patch, tonic firing in AAV-Scr and AAV-shANO2 TC neurons were generated by 140 pA current injection. (l) An analysis of adaptation index in both AAV-Scr-infected (n=6 from three mice) and AAV-shANO2-infected (n=6 from four mice) TC neurons showed that AAV-shANO2 infection induced a significant decrease in spike-frequency adaptation (*P=0.028, t-test). (m) Tonic firing was generated by current injection in CA1 hippocampal pyramidal neurons infected with AAV-Scr or AAV-shANO2.(n) Adaptation indexes were averaged in both AAV-Scr-infected (n=9 from three mice) and AAV-shANO2-infected (n=8 from three mice) neurons (**P=0.0098, t-test). (o) Single action potentials from AAV-Scr (black) and AAV-shANO2 (red) infected CA1 neurons. (p) There was no difference in the half-width of single action potentials between AAV-Scr-infected (black, n=9 from three mice) and AAV-shANO2-infected (red, n=8 from three mice) CA1 neurons (P=0.11, t-test). Error bars represent mean±s.e.