Extended Data Fig. 6: Bilateral dLight measurement and mutant FLIM-AKAR control experiments.

a, Schematic describing a strategy to measure DA level in both hemispheres. AAV9-hSyn-dLight1.1 was bilaterally injected into the NAc of C57BL/6J mice. Then, two optical fibres were implanted 200 μm above the injection sites in two hemispheres. b, Relationship between dLight signals from two hemispheres. Left, average correlation between two dLight signals (Δf/f) during trial duration (−5 to 15 s with respect to the trigger zone entry) and all time (analysed in 20-s time window for each linear fit) plotted as mean ± 95% confidence interval across mouse averages (two-sided paired t-test, P = 0.002). Middle, correlation of individual mouse for trial duration plotted as mean ± 95% confidence interval across session averages for each mouse. Right, correlation of individual mouse for all time plotted as in middle. n = 7 mice. c, Comparison between FLIM-AKAR and FLIM-AKAR(T391A), which has a point mutation at the PKA phosphorylation site, signals. AAV1-FLEX-FLIM-AKAR or AAV1-FLEX-FLIM- FLIM-AKAR(T391A) was injected into the NAc of Drd1a-cre or Adora2a-cre mice for these experiments. From the top, D1R-SPN FLIM-AKAR (D1R-SPN AKAR), D1R-SPN FLIM-AKAR(T391A) (D1R-SPN mAKAR), D2R-SPN FLIM-AKAR (D2R-SPN AKAR), D2R-SPN FLIM-AKAR(T391A) (D2R-SPN mAKAR). From the left, signals were aligned to the time (dashed vertical line) of ‘LED on’ for success and failure trials separately, ‘receptacle entry’ for success trials and ‘pellet dispensing’ for success, reward-omission and LED-omission trials. Signals for beginner, intermediate, expert, reward-omission (of expert mice) and rewarded LED-omission (of expert mice) trials are shown in red, orange, green, blue and purple, respectively. Plotted as mean ± s.e.m. across mice. n = 14 mice (D1R-SPN AKAR), 7 mice (D1R-SPN mAKAR), 18 mice (D2R-SPN AKAR), 6 mice (D2R-SPN mAKAR).