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  • An expanded palette of dopa...
    Patriarchi, Tommaso; Mohebi, Ali; Sun, Junqing; Marley, Aaron; Liang, Ruqiang; Dong, Chunyang; Puhger, Kyle; Mizuno, Grace Or; Davis, Carolyn M; Wiltgen, Brian; von Zastrow, Mark; Berke, Joshua D; Tian, Lin

    Nature methods, 11/2020, Letnik: 17, Številka: 11
    Journal Article

    Genetically encoded dopamine sensors based on green fluorescent protein (GFP) enable high-resolution imaging of dopamine dynamics in behaving animals. However, these GFP-based variants cannot be readily combined with commonly used optical sensors and actuators, due to spectral overlap. We therefore engineered red-shifted variants of dopamine sensors called RdLight1, based on mApple. RdLight1 can be combined with GFP-based sensors with minimal interference and shows high photostability, permitting prolonged continuous imaging. We demonstrate the utility of RdLight1 for receptor-specific pharmacological analysis in cell culture, simultaneous assessment of dopamine release and cell-type-specific neuronal activity and simultaneous subsecond monitoring of multiple neurotransmitters in freely behaving rats. Dual-color photometry revealed that dopamine release in the nucleus accumbens evoked by reward-predictive cues is accompanied by a rapid suppression of glutamate release. By enabling multiplexed imaging of dopamine with other circuit components in vivo, RdLight1 opens avenues for understanding many aspects of dopamine biology.