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In Vivo Calcium Imaging in C. elegans Body Wall Muscles
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Simultaneous optogenetic manipulation and calcium imaging in freely moving C. elegans
Frederick B Shipley1, Christopher M Clark2, Mark J Alkema2
1Lewis Sigler Institute for Integrative Genomics, Princeton University Princeton, NJ, USA.
Frontiers in Neural Circuits
|April 10, 2014
Summary
This study introduces a new instrument for observing and controlling neural activity in freely moving Caenorhabditis elegans (worms). The technology allows researchers to link specific neural circuit dynamics to animal behavior in real-time.
Area of Science:
- Neuroscience
- Biophysics
- Genetics
Background:
- Understanding neural circuits requires observing neural activity during natural behaviors.
- Current methods often limit simultaneous recording and manipulation in freely moving organisms.
- The nematode Caenorhabditis elegans is a powerful model for studying neural control of behavior.
Purpose of the Study:
- To develop and validate an instrument for simultaneous optical monitoring and manipulation of neural activity in freely moving Caenorhabditis elegans.
- To investigate the relationship between sensory neuron activation, interneuron dynamics, and locomotion in C. elegans.
- To enable real-time, closed-loop control of neural circuits based on observed behavior.
Main Methods:
- Utilized optical recording of neural activity via GCaMP3 calcium indicator.
- Employed optogenetic manipulation using Channelrhodopsin.
- Developed real-time computer vision software for behavioral tracking and neuron targeting.
- Implemented a Digital Micromirror Device (DMD) for patterned illumination and selective neural stimulation.
- Integrated behavioral tracking with neural manipulation for closed-loop experiments.
Main Results:
- Successfully recorded and manipulated neural activity in freely moving C. elegans.
- Demonstrated real-time tracking of worm behavior and dynamic adjustment of neural stimulation patterns.
- Observed calcium transients in backward locomotion command interneurons (AVA) in response to optogenetic activation of mechanosensory neurons (ALM, AVM).
- Provided direct evidence linking specific sensory inputs to interneuron activity and motor outputs.
Conclusions:
- The developed instrument enables unprecedented simultaneous recording and manipulation of neural activity during unrestrained behavior in C. elegans.
- This technology facilitates detailed investigation of neural circuit function and its role in behavior.
- Future applications include dissecting complex neural circuits and understanding sensory-motor transformations.
