Extended Data Fig. 6: Tone detection behaviour is compromised by locomotion, is auditory-cortex dependent, and adapts following VR experience.

a, Data points show mean and s.e. detection rates for N = 4 mice as a function of tone intensity for trials performed during rest with infusion of either saline (black) or muscimol (magenta) into the auditory cortex. b, Difference in performance as a function of intensity for each mouse (grey dots). Large connected dots show mean difference in performance and coloured dots indicate intensities at which performance was significantly different (P < 0.05) across conditions (N = 19 mice, repeated measures two-way ANOVA followed by post-hoc Tukey test). c, Tone-evoked responses from putative excitatory neurons recorded from VGAT::ChR2 mouse without (black) and with (blue) simultaneous blue laser stimulation. Optogenetic activation of inhibitory neurons decreases the spontaneous and tone-evoked firing rates of excitatory neurons. n = 23 neurons, N = 1 mouse. d, Tone-evoked firing rates during rest are weaker during optogenetic activation of inhibitory interneurons. Dashed line is unity. (n = 23 neurons, N = 1 mouse; P < 0.05, two-sided paired t-test.) e, Tone detection performance (N = 6 mice) during rest (black) and rest with optogenetic activation of auditory cortical inhibitory neurons (blue). Mice were worse at detecting tones on optogenetic trials (repeated measures two-way ANOVA, factors: intensity × laser state, P(intensity × laser state) = 0.0028, F(2, 10) = 11.23, post-hoc Tukey test at individual intensities, blue asterisk, P < 0.05 on laser trials) compared to rest. f, Tone detection performance (N = 6 mice) during rest (black) and rest with optogenetic activation of M2 terminals in auditory cortex (blue). Four of these mice were presented with 8-kHz tones and the remaining two were presented with 4-kHz tones. Mice were worse at detecting tones on optogenetic trials regardless of the tone frequency. (Statistics similar to e, P(intensity × laser state) = 0.01, F(2, 10) = 6.66, blue asterisk, P < 0.05 on laser trials). g, Average psychometric functions (N = 3 mice) showing detection rates as a function of tone intensity for trials performed during rest when visual cortex was inhibited. (repeated measures two-way ANOVA, P(intensity × laser state) = 0.33, F(2, 4) = 1.47). h, Average psychometric functions (N = 2 mice) showing detection rates as a function of tone intensity for trials performed during rest (black) and during rest with laser stimulation (blue) by mice injected with an AAV encoding eGFP in M2. These controls show that laser stimulation of auditory cortex in the absence of ChR2 does not influence behaviour. i, Average psychometric functions (N = 8 mice) showing detection rates as a function of tone intensity for trials performed during rest (black) and during rest with laser stimulation (blue) when the optical fibre was placed over intact skull near, but not directly over auditory cortex. Five of eight mice were injected with an AAV encoding ChR2 into M2, of which three were presented with 8-kHz tones and 2 with 4-kHz tones. The other three were VGAT::ChR2 mice presented with 8-kHz tones. These controls show that sham laser stimulation (which is visible to the mouse) alone improves behaviour (repeated measures two-way ANOVA, factors: intensity × laser state, P(interaction) = 0.0066, F(2, 14) = 7.35, post-hoc Tukey tests, blue asterisk, P < 0.05). j, Difference in hit rates in response to tone A relative to tone B during rest before (pre) and after (post) aVR experience with tone A.). Lines represent mean difference and shaded regions show s.e. for N = 10 mice. There is no difference in rest performance before and after aVR experience. (repeated measures two-way ANOVA in each panel, factors: intensity × time of testing, P(time of testing) = 0.46, F(1, 9) = 0.61). k, Difference in hit rates in response to tone A relative to tone B during running before (pre) and after (post) aVR experience with tone A. Lines represent mean difference and shaded regions show s.e. for N = 10 mice. Mice are significantly better at detecting tone B than tone A after aVR experience, indicating that this is a movement-specific change (repeated measures two-way ANOVA in each panel, factors: intensity × time of testing, P(time of testing) = 0.04, F(1, 9) = 8.07, red asterisk, P < 0.05, p values in j, k corrected using the Holm–Bonferroni method. For further statistical details, see Supplementary Table 1.