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Imaging calcium waves in cerebellar Bergmann glia
Cold Spring Harbor Protocols
|January 4, 2013
Summary
This study details a method to record calcium (Ca2+) waves in Bergmann glia (BG) cells within the cerebellum. This protocol allows for reliable, repeated measurements essential for understanding astrocyte signaling in the brain.
Area of Science:
- Neuroscience
- Cell Biology
- Astrocyte Biology
Background:
- Bergmann glia (BG) are large radial astrocytes in the cerebellum.
- Their extensive processes ensheathe thousands of parallel fiber (PF) synapses.
- Stable calcium (Ca2+) measurements in astrocytes are challenging due to thin processes.
Purpose of the Study:
- To describe a reliable protocol for recording synaptically evoked Ca2+ waves in individual BG.
- To enable quantitative studies of Ca2+ signaling in astrocytes.
- To characterize mechanisms and functional consequences of Ca2+ increases in astrocytes.
Main Methods:
- Recording synaptically evoked Ca2+ waves from individual BG in cerebellar slices.
- Using a cooled charge-coupled device (CCD) camera for fluorescence measurements.
- Loading BG with high-affinity Ca2+ indicators like Fura-2 for up to 2 hours.
Main Results:
- Stimulation of PF synapses reliably triggers long-lasting Ca2+ responses in BG processes.
- BG Ca2+ waves can be repeatedly evoked due to rare spontaneous transients.
- CCD technology provides a straightforward method for Ca2+ wave characterization.
Conclusions:
- This protocol facilitates the study of Ca2+ signaling in Bergmann glia.
- It enables quantitative analysis of astrocyte-mediated Ca2+ dynamics.
- Provides insights into the functional roles of Ca2+ waves in cerebellar astrocytes.
