Fig. 5: Chemogenetic inhibition of ChCs disrupts the direction selectivity of premotor neurons.
From: An adaptive behavioral control motif mediated by cortical axo-axonic inhibition
a, In vivo two-photon Ca2+ imaging of ChC and neighboring neurons expressing GCaMP6s and selectively co-expressing the chemogenetic silencer hM4Di-mCherry in ChCs of M2, in an FlpO-dependent manner, using Nkx2.1-2a-CreER::Flex-FlpO mice (ChC-Flp mice, n = 9). b, Bath-application of CNO (10 μM) reduced the firing rate in ChCs expressing hM4Di. Example whole-cell current-clamp recording from a ChC. c, The average number of successes of ChC-hM4Di mice increased with training from sessions 1 to 7. CNO or saline injection was followed on session 8 or 9 (n = 9 mice, one-way repeated-measures ANOVA, Fsession = 31.1, P = 3.09 × 10−6; Fisher multiple comparisons tests, session 7 versus CNO, P = 2.40 × 10−6; session CNO versus saline, P = 7.58 × 10−6; session 7 versus saline, P = 0.53). d, Cumulative turning angle of ChC-hM4Di mice across conditions (session 7, CNO and saline). In the box plot, the midline, box size and whisker indicate median, 25th–75th percentiles and 10th–90th percentiles, respectively. e, Comparison of cumulative turning angle across conditions (one-way repeated-measures ANOVA, Fsession = 8.99, P = 0.0024; Fisher multiple comparisons tests, session 7 versus CNO, P = 0.0035; session 7 versus saline, P = 0.45; CNO versus saline, P = 0.0013). f, Comparison of performance between conditions (n = 6 mice for ChC control with saline or CNO; n = 9 mice for ChC-hM4Di with saline or CNO, one-way ANOVA, Fsession = 9.50, P = 2.06 × 10−4). In the box plot, the midline, square, box size and whisker indicate median, mean, 25th–75th percentiles and 10th–90th percentiles, respectively. g, Example ΔF/F traces of a ChC during locomotion in CNO and saline conditions. h, Average ΔF/F traces of ChCs aligned to movement onset in CNO and saline conditions (six ChC-hM4Di mice, n = 497 cells for CNO; n = 475 cells for saline). i, Co-activity percentage of neurons during periods of locomotion and rest (two-tailed Friedman test, χ2 = 9.33, P = 0.0094 for locomotion; χ2 = 0.33, P = 0.85 for rest). j, Example tuning curves of each individual premotor neuron for MD in CNO (top) and saline (bottom) conditions. Data are sorted from the location of peak likelihood probability P(active|MD). Corresponding precisions of population responses for each MD (right). k, Average precision curves of population responses across later learning, CNO and saline conditions. l, Comparison of population response precisions (one-way repeated-measures ANOVA, Fsession = 6.39, P = 0.016; Fisher multiple comparisons tests, session 7 versus CNO, P = 0.0068; session CNO versus saline, P = 0.023; session 7 versus saline, P = 0.486). m, Normalized percentage of active cells in the population as a function of distance from the PD (n = 484 cells for session 7, n = 407 cells for CNO, n = 582 cells for saline). n, Changes in the percentage of active cells across conditions (one-way repeated-measures ANOVA with Greenhouse–Geisser correction, Fsession = 7.26, P = 0.037; Fisher multiple comparisons tests, session 7 versus CNO, P = 0.006; session CNO versus saline, P = 0.011; session 7 versus saline, P = 0.372). o, Pairwise correlations with respect to ΔPD normalized by session 7 (angular difference in PD between neuronal pairs, n = 17,933 pairs for session 7; n = 24,205 pairs for CNO; n = 22,735 pairs for saline). *P < 0.05; **P < 0.01; ***P < 0.001; NS, not significant; CW, clockwise; CCW, counterclockwise. Error bars and shading indicate s.e.m.
