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Imaging neuromodulators with high spatiotemporal resolution using genetically encoded indicators.

Tommaso Patriarchi^1,2, Jounhong Ryan Cho³, Katharina Merten⁴

¹Department of Biochemistry and Molecular Medicine, University of California, Davis, Davis, CA, USA.

Nature Protocols

|November 17, 2019

Summary

This summary is machine-generated.

Researchers engineered dLight1, a novel genetically encoded dopamine (DA) sensor. This tool enables precise in vivo measurement of DA release, advancing neuroscience research.

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Area of Science:

Neuroscience
Molecular Biology
Biotechnology

Background:

Neural activity, including neurotransmitter release, is crucial for brain function and behavior.
Genetically encoded sensors offer high spatial and temporal resolution for measuring neural signaling.
Dopamine (DA) signaling plays a vital role in various brain functions.

Purpose of the Study:

To engineer, characterize, and validate dLight1, a new genetically encoded dopamine sensor.
To provide a detailed protocol for assessing sensor properties and in vivo application.
To establish dLight1 as a benchmark for developing novel neuromodulator sensors.

Main Methods:

Engineering of dLight1 based on human inert dopamine receptors.
Characterization of sensor properties: dynamic range, affinity, specificity, kinetics, and interaction with endogenous signaling.
Application of dLight1 in cultured cells, brain slices, and behaving mice using fiber photometry and two-photon imaging.

Main Results:

dLight1 demonstrates high molecular specificity, appropriate affinity and kinetics, and excellent sensitivity for in vivo dopamine detection.
The protocol allows systematic validation of sensor performance across different biological systems.
Successful measurement of dLight1 transients in vivo using established imaging techniques.

Conclusions:

dLight1 is a robust and sensitive tool for measuring dopamine release in vivo.
The provided protocol facilitates the development and utilization of novel neuromodulator sensors.
This work advances the ability to study neural circuits and behavior.