Bioluminescence Imaging of Neuronal Network Dynamics Using Aequorin-Based Calcium Sensors

Area of Science:

• Neuroscience
• Biotechnology
• Molecular Imaging

Background:

• Optogenetic tools are crucial for real-time neuronal activity imaging.
• Bioluminescent probes offer advantages over fluorescent sensors, including no need for external illumination and minimal background noise.
• Aequorin-based probes are suitable for long-duration, simultaneous recordings of multiple neurons due to low cytotoxicity and wide dynamic range.

Purpose of the Study:

• To describe a protocol for monitoring and analyzing neuronal ensemble dynamics.
• To demonstrate the utility of whole-field bioluminescence imaging with an aequorin-based sensor.
• To provide a method for studying neural circuit activity in brain slices.

Main Methods:

• Engineering bioluminescent probes from the natural calcium sensor aequorin.
• Utilizing whole-field bioluminescence imaging for neuronal activity detection.
• Developing protocols for monitoring and analyzing neuronal ensemble dynamics in brain slices.

Main Results:

• Demonstrated successful application of aequorin-based bioluminescent probes for neuronal activity imaging.
• Showcased the effectiveness of whole-field bioluminescence imaging for capturing dynamics of neuronal ensembles.
• Validated the protocol for analyzing neural circuit activity in brain slice preparations.

Conclusions:

• Aequorin-based bioluminescent imaging provides a powerful, illumination-free method for studying neuronal ensemble dynamics.
• The described protocol enables long-duration, whole-field recordings of neural activity in brain slices.
• This approach enhances the study of neural circuits and network function.