Next-generation GRAB sensors for monitoring dopaminergic activity in vivo

Nat Methods. 2020 Nov;17(11):1156-1166. doi: 10.1038/s41592-020-00981-9. Epub 2020 Oct 21.


Dopamine (DA) plays a critical role in the brain, and the ability to directly measure dopaminergic activity is essential for understanding its physiological functions. We therefore developed red fluorescent G-protein-coupled receptor-activation-based DA (GRABDA) sensors and optimized versions of green fluorescent GRABDA sensors. In response to extracellular DA, both the red and green GRABDA sensors exhibit a large increase in fluorescence, with subcellular resolution, subsecond kinetics and nanomolar-to-submicromolar affinity. Moreover, the GRABDA sensors resolve evoked DA release in mouse brain slices, detect evoked compartmental DA release from a single neuron in live flies and report optogenetically elicited nigrostriatal DA release as well as mesoaccumbens dopaminergic activity during sexual behavior in freely behaving mice. Coexpressing red GRABDA with either green GRABDA or the calcium indicator GCaMP6s allows tracking of dopaminergic signaling and neuronal activity in distinct circuits in vivo.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Brain / metabolism*
  • Dopamine / metabolism*
  • Drosophila / genetics
  • Drosophila / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Neurons / metabolism
  • Rats
  • Receptors, Dopamine / genetics
  • Receptors, Dopamine / metabolism
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*
  • Red Fluorescent Protein
  • Sexual Behavior / physiology*


  • Luminescent Proteins
  • Receptors, Dopamine
  • Receptors, G-Protein-Coupled
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Dopamine