High-throughput recording in mice performing a visual Go/No-Go task.

(A) Schematic of the task. (B) Average correct rate during training (mean ± SEM, n = 7 mice). (C) Photos showing the uFINE-M shanks and recording sites. (D) Schematic showing the implantation sites of uFINE-M arrays, along with example single-unit waveforms recorded from each brain region. Brain section images are adapted from Allen reference atlas23. (E) Example spike rasters during two trials. (F) Top, the number of single units recorded in each brain region. Each data point represents data from an individual recording session. Bottom, the total number of single units recorded during training (n = 5 mice). Each symbol represents data from an individual mouse.

Activity changes throughout task learning.

(A) Averaged firing rate aligned to the visual stimulus onset for all CR trials and Hit trials in the early and expert stages (n = 118 early CR and 828 early Hit trials from 7 sessions of 3 mice, 610 expert CR trials and 677 expert Hit trials from the same mice). Shading, SEM. (B) Left, distribution of activity onset timing across time. Right, activity onset timing of each region. Each data point represents data from a neuron. (C) Same as B but for Hit trials. (D) Comparison of activity onset timing between the early and expert stages. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, two-way ANOVA, Sidak’s multiple comparison. Error bars, SEM.

Definition of functional connection.

(A) Schematic of data processing flow of calculating functional connectivity. For each 200-ms time window (t), cross-correlation scores were calculated between spike trains of neuron pairs and the percentage of neuron pairs that showed significant cross-correlations was treated as functional connection strength between brain regions. The regional connection matrix was then ranked from 1 to 10 to evaluate the relative importance of regional connection compared with other connections within the same time window of the same trial. (B) Details of processing stages. (C) Connection rank matrix calculated from the example data in A.

Ranking dynamics in CR trials during learning.

(A) Input/output ranking dynamics during early and expert CR trials. (B) Average input rank of each brain region in the early stimulus period (0–400 ms after stimulus onset), late stimulus period (400– 800 ms after stimulus onset), early response period (800–1800 ms after stimulus onset), and late response period (1800–2800 ms after stimulus onset) of early and expert CR trials, mapped on brain atlas 28. ΔRank represents the rank change between the expert and early stages. (C) Same as B but for output ranks. n = 118 early CR trials from 7 sessions of 3 mice, and 610 expert CR trials from 6 sessions of same mice. Error bars, SEM.

Ranking dynamics in Hit trials during learning.

(A) Input/output ranking dynamics in early and expert Hit trials. (B) Average input rank of each brain region in the early stimulus period (0–400 ms after stimulus onset), late stimulus period (400– 800 ms after stimulus onset), early response period (800–1800 ms after stimulus onset), and late response period (1800–2800 ms after stimulus onset) of early and expert Hit trials, mapped on brain atlas 28. ΔRank represents the rank change between the expert and early stages. (C) Same as B but for output ranks. n = 828 early Hit trials from 7 sessions of 3 mice, and 677 expert Hit trials from 6 sessions of 3 mice. Error bars, SEM.

Rank increase in CR trials was attributed to elevated input/output rank from/to other regions.

(A) Average input rank changes for the four regions (V1, V2M, M2, and OFC) that showed increased rank values in the stimulus or response period of CR trials during visual learning. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, two-way ANOVA, Sidak’s multiple comparison. (B) Same as A but for the response period of CR trials. (C-D) Same as A-B but for output ranks. n = 118 early CR trials from 7 sessions of 3 mice, and 610 expert CR trials from 6 sessions of 3 mice. Dashed lines at rank 5 indicate the average level of random data. Error bars, SEM. Stim, stimulus period. Res, response period.

Encoding of visual stimulus information during task learning.

(A) Schematic of the ROC analyses and example data from a neuron preferring the No-Go stimulus. (B) Percentage of stimulus-selective neurons in each brain region during the early and late training stages. (C) Mean percentage of stimulus-selective neurons in the early (0–400 ms after stimulus onset) and late (400–800 ms after stimulus onset) stimulus periods. n = 10 time bins for the early training stage and 30 time bins for the expert training stage. (D) Same as C, but for the early (800–1800 ms after stimulus onset), and late (1800–2800 ms after stimulus onset) response periods. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, two-way ANOVA, Sidak’s multiple comparison. n = 34 time bins for the early training stage and 102 time bins for late training stage. (E) Correlation between stimulus encoding peak time and input/output rank in expert CR trials. A significant correlation was observed during the stimulus period but not the response period (Pearson’s correlation).

The effects of bilateral optogenetic inhibition on task performance.

(A) Expression of AAV2/9-mCaMKIIa-eJaws3.0-mRuby3-WPRE-pA in the OFC. VO: ventral orbitofrontal cortex; MO: medial orbitofrontal cortex. Regions were named according to the Paxinos atlas 35. (B) Correct rejection rate for the OFC-stimulus period inhibition group (eJaws 3.0 Stim), and the control group (mCherry Stim). Shading, SEM. n = 8 and 14 mice for the eJaws 3.0 and mCherry group, respectively. ****p < 0.0001, significance for the group factor in two-way ANOVA. (C) Same as B, but for the OFC-response period inhibition group (n = 8 mice) and control group (n = 16 mice). (D) Average miss rate for each mouse in the OFC manipulation group and control group. (E-H): Same as A to D but for V2M inhibition. n = 8 mice for each manipulation group. ***p < 0.001, significance for the group factor in two-way ANOVA. The control group here was the same group of mice in A-D.