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Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging.
Shai Berlin1,2, Elizabeth C Carroll1, Zachary L Newman1
1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
Nature Methods
|July 14, 2015
Summary
Researchers developed a novel photoactivatable calcium indicator for simultaneous visualization of neuronal structure and activity. This tool enables precise circuit mapping by allowing single-cell selection within dense populations for detailed analysis.
Area of Science:
- Neuroscience
- Molecular Biology
- Biotechnology
Background:
- Understanding neural circuits requires integrating structural and functional connectivity data.
- Existing optical tools often assess either morphology or activity, but rarely both simultaneously.
- A need exists for integrated probes that combine high-contrast photolabeling with sensitive calcium detection.
Purpose of the Study:
- To develop a single-color protein sensor integrating photolabeling and calcium sensing.
- To enable selective visualization and activity measurement of individual neurons within complex networks.
- To advance circuit mapping by combining structural and functional connectivity assessments.
Main Methods:
- Generation of photoactivatable genetically encoded calcium indicators.
- Utilized a single-color protein sensor design combining photolabeling and calcium detection.
- Demonstrated in cultured neurons, fruit fly, and zebrafish larvae models.
Main Results:
- Successfully visualized morphology and measured activity, synaptic transmission, and connectivity of selected single cells.
- Achieved high-contrast photolabeling and high-sensitivity calcium detection within a single sensor.
- Enabled selection of individual cells from dense populations for detailed analysis.
Conclusions:
- The developed photoactivatable calcium indicators provide a powerful tool for integrated circuit mapping.
- This technology facilitates detailed analysis of neuronal structure and function in complex biological systems.
- The design strategy is adaptable for other sensors based on circularly permutated GFP (cpGFP).
