Fig. 5: Progressive engagement of AHN^Vgat+ neurons on EPM.

a–e Recording of AHN^Vgat+ GCaMP6s signals on EPM. a Average ΔF/F values in EPM open and closed arm for the first trial with the a heatmap representation of EPM ΔF/F value in an example (right). n = 13 mice. Two-tailed paired t test p = 0.048. b, c Average open-arm time (b) and ΔF/F values (c) in the first or second trial. n = 11 mice. Open-arm time, Wilcoxon matched-pairs signed rank test, p = 0.01; ΔF/F, two-tailed paired t test, p = 0.006. d Average ΔF/F values in the first or second 5 min of the first trial. n = 12 mice. e Correlation between average open-arm ΔF/F value and the time spent in the closed arm. n = 14 mice. Pearson correlation. f–h Single-unit recordings of AHN neurons. f Raster plot (top) and the average (bottom) of the firing rates of an example single-unit aligned to the onset of the object approach (left) and entering the EPM open-arm (right). g Heatmap representation of the normalized single-unit responses in Z scores sorted by response magnitude aligned to behavioral onset. n = 98 units from 3 mice. h Quantification of the number of single-units in each response category. i–l GtACR1-mediated optogenetic inhibition of AHN^Vgat+ neurons on EPM. i Left, schematics showing light delivery restricted to a random open-arm; right, example trajectories from an EYFP or a GtACR1 animal as indicated. j, k Time spent in EPM open-arm. l Velocity on EPM open-arm. n = 7 EYFP and 7 GtACR1 males. j. EYFP, Wilcoxon matched-pairs signed rank test, p = 0.69, GtACR1, two-tailed paired t test, p = 0.003. k. Two-way repeated measures ANOVA followed by Sidak’s multiple comparison test. *p < 0.05; **p < 0.01. Data are presented as mean values ±SEM.