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An expanded palette of dopamine sensors for multiplex imaging in vivo.

  • Author(s): 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
  • et al.
Abstract

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.

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