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Ratiometric Calcium Imaging of Individual Neurons in Behaving Caenorhabditis Elegans

Ratiometric Calcium Imaging of Individual Neurons in Behaving Caenorhabditis Elegans

Published on: February 7, 2018

Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans.

Jun Liu¹, Elsa Bonnard^1,2, Monika Scholz¹

¹Max Planck Research Group Neural Information Flow, Max Planck Institute for Neurobiology of Behavior-caesar, Bonn 53175, Germany.

|July 29, 2024

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We optimized a new calcium indicator, GCaMP8f, for use in C. elegans. This genetically encoded calcium indicator (GECI) offers improved brightness and speed for studying muscle and neuron activity.

Area of Science:

Neuroscience
Molecular Biology
Genetics

Background:

Genetically encoded calcium indicators (GECIs) are crucial for observing cellular activity in real-time.
Ultrafast and highly sensitive GECIs are needed for precise measurements of dynamic biological processes.
Previous studies demonstrated novel GECIs (jGCaMP8) in mammals and Drosophila, but not in Caenorhabditis elegans.

Purpose of the Study:

To develop and validate an optimized GCaMP8f construct for C. elegans.
To compare the performance of GCaMP8f against GCaMP7f in C. elegans pharyngeal muscle.
To assess the utility of GCaMP8f for detecting neuronal activity in C. elegans.

Main Methods:

Constructing and generating C. elegans strains expressing GCaMP8f under the myo-2 promoter.

Keywords:

GCaMP calcium imaging genetically encoded indicator neuronal activity pharyngeal muscle

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In vivo Neuronal Calcium Imaging in C. elegans

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Comparing GCaMP8f and GCaMP7f in pharyngeal muscle activity measurements.

Validating GCaMP8f in touch receptor neurons to detect calcium transients.

Main Results:

GCaMP8f demonstrated superior brightness and faster kinetics compared to GCaMP7f in C. elegans pharyngeal muscle.
GCaMP8f did not disrupt the intrinsic contraction dynamics of the pharynx.
GCaMP8f successfully detected robust calcium transients in touch receptor neurons, even with small stimuli.

Conclusions:

GCaMP8f is a powerful new tool for C. elegans research, enabling the study of fast calcium dynamics.
This indicator allows for high-resolution imaging across multiple cell types at low magnifications.
GCaMP8f significantly advances the capability to investigate intracellular dynamics in C. elegans.