12/21/2023 0 Comments Dlight dopamine sensor![]() 2 : The sequences of GRABDA sensors and the residues related to affinity-tuning, cpRFP and cpEGFP optimization.Įxtended Data Fig. 1 : The development of red fluorescent DA sensors and second-generation green fluorescent DA sensors.Įxtended Data Fig. 6 : GRABDA sensors can be used to measure dopaminergic activity in the mouse NAc during sexual behavior.Įxtended Data Fig. 5 : GRABDA sensors can detect optogenetically induced nigrostriatal DA release in freely moving mice.įig. 4 : In vivo two-photon imaging of DA dynamics in Drosophila using GRABDA sensors.įig. 3 : GRABDA sensors can be used to measure DA release in acute mouse brain slices.įig. 2 : Characterization of GRABDA sensors in HEK293T cells and cultured rat cortical neurons.įig. 1 : Development of red fluorescent DA sensors and second-generation green fluorescent DA sensors.įig. Coexpressing red GRAB DA with either green GRAB DA or the calcium indicator GCaMP6s allows tracking of dopaminergic signaling and neuronal activity in distinct circuits in vivo. Moreover, the GRAB DA 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. In response to extracellular DA, both the red and green GRAB DA sensors exhibit a large increase in fluorescence, with subcellular resolution, subsecond kinetics and nanomolar-to-submicromolar affinity. We therefore developed red fluorescent G-protein-coupled receptor-activation-based DA (GRAB DA) sensors and optimized versions of green fluorescent GRAB DA sensors. Altogether this review should act as a tool to guide DA sensor choice for end-users.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 then outline a map of DA heterogeneity across the brain and provide a guide for optimal sensor choice and implementation based on local DA levels and other experimental parameters. In this review, we use DA as an example we briefly summarize old and new techniques to monitor DA release, including DA biosensors. Molecular specificity, sensor kinetics, spectral properties, brightness, sensor scaffold and pharmacology can further influence sensor choice depending on the experimental question. Sensor properties, most importantly their affinity and dynamic range, must be carefully chosen to match local DA levels. When implementing these tools in the laboratory, it is important to consider there is not a ‘one-size-fits-all’ sensor. Combined with rapid developments in in vivo imaging, these sensors have the potential to transform the field of DA sensing and DA-based drug discovery. Recently, red and green genetically encoded sensors for DA (dLight, GRAB-DA) were developed and now provide the ability to track release dynamics at a subsecond resolution, with submicromolar affinity and high molecular specificity. Understanding how dopamine (DA) encodes behavior depends on technologies that can reliably monitor DA release in freely-behaving animals.
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