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Studying Axon-Astrocyte Functional Interactions by 3D Two-Photon Ca2+ Imaging: A Practical Guide to Experiments and
Iaroslav Savtchouk1, Giovanni Carriero1, Andrea Volterra1
1Department of Fundamental Neurosciences, University of Lausanne, Lausanne, Switzerland.
Frontiers in Cellular Neuroscience
|May 1, 2018
Summary
This study presents a protocol and software for analyzing 3D calcium (Ca2+) dynamics in astrocytes. The tools enable interactive exploration and correlation analysis of astrocytic Ca2+ transients, crucial for understanding brain function.
Area of Science:
- Neuroscience
- Cellular Neuroscience
- Neuroimaging
Background:
- Fast volumetric imaging generates large functional datasets.
- Existing analysis tools are suboptimal for astrocytic Ca2+ dynamics due to unpredictable patterns.
Purpose of the Study:
- To provide a protocol for recording and analyzing 3D Ca2+ transients in GCaMP6f-expressing astrocytes.
- To introduce novel software tools for interactive exploration, filtering, and time-correlation analysis of astrocytic Ca2+ signals.
- To discuss parameters for axonal stimulation/response experiments in various brain regions.
Main Methods:
- Utilized GCaMP6f-expressing astrocytes in adult brain slices.
- Employed axonal stimulation to evoke Ca2+ transients.
- Developed and applied custom Image J plugins (MultiROI_TZ_profiler, Gaussian_Filter5D, Correlation_Calculator) for data analysis.
Main Results:
- Demonstrated a method for recording and analyzing 3D Ca2+ transients in astrocytes.
- Provided interactive tools for exploring, filtering, and correlating Ca2+ dynamics.
- Released software plugins for Image J to facilitate astrocytic Ca2+ imaging analysis.
Conclusions:
- The developed protocol and software facilitate the analysis of astrocytic Ca2+ signaling.
- These tools address the limitations of existing frameworks for analyzing complex astrocytic Ca2+ transients.
- The findings aid in understanding the role of astrocytes in neural circuits through Ca2+ dynamics.