Extended Data Fig. 5: Movement and optical artefacts cannot explain dLight and jRCaMP signal patterns.
From: Cell-type-specific asynchronous modulation of PKA by dopamine in learning
a, Īf/f (%) of different controls. The average 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. Dashed vertical lines indicate the behavioural time stamps (TĀ =Ā trigger zone entry, LĀ =Ā LED on, ZĀ =Ā receptacle zone entry, DĀ =Ā pellet dispensing, RĀ =Ā receptacle entry). Top, Īf/f (%) of eGFP signal from the VTA of DAT-IRES-cre mice (nĀ =Ā 4 mice) that were injected with AAV1-Cag-FLEX-eGFP into the VTA. Middle, Īf/f (%) of eGFP signal from the NAc of DAT-IRES-cre mice (nĀ =Ā 4 mice) that were injected with AAV1-Cag-FLEX-eGFP into the VTA. Bottom, Īf/f (%) of DA-binding mutant dLight (D103A mutation) signal from the NAc of C57BL/6J mice (nĀ =Ā 8 mice) that were injected with AAV9-hSyn-dLightD103A into the NAc. b, Īf/f (%) of different controls that are magnified in Īf/f axis and demagnified in time axis. VTA eGFP (left, nĀ =Ā 4 mice), NAc eGFP (middle, nĀ =Ā 4 mice) and NAc mutant dLight (right, nĀ =Ā 8 mice) signal aligned to the time of trigger zone entry (dashed vertical line). There was a minor (compared to sensor responses) change in NAc eGFP and mutant dLight signal that develops across learning (possibly owing to haemodynamic effects). c, Test for the optical crosstalk between green and red spectrum for simultaneous dual-colour photometry for dLight and jRCaMP. Mice were given unexpected free food pellets, and signal was aligned to the time of pellet dispensing. Left, raw fluorescence signal in red and green spectrum from NAc of C57BL/6J mice (nĀ =Ā 3 mice, 10 trials prt mouse) injected with AAV9-hSyn-dLight1.1 into the NAc. Right, raw fluorescence signal in red and green spectrum from the NAc of DAT-IRES-cre mice (nĀ =Ā 3 mice, 10 trials per mouse) injected with AAV1-hSyn-FLEX-NES-jRCaMP1b into the VTA. d, Baseline (pretrial) raw fluorescence estimating the change in a signal strength due to photobleaching and viral expression change across days. Raw fluorescence was normalized by the maximum value of each mouse across all sessions. All graphs are plotted as meanĀ Ā±Ā s.e.m. across mice.
