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Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans. | LitMetric

Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans.

Genetics

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

Published: October 2024

Improved genetically encoded calcium indicators (GECIs) are essential for capturing intracellular dynamics of both muscle and neurons. A novel set of GECIs with ultrafast kinetics and high sensitivity was recently reported by Zhang et al. (2023). While these indicators, called jGCaMP8, were demonstrated to work in Drosophila and mice, data for Caenorhabditis elegans were not reported. Here, we present an optimized construct for C. elegans and use this to generate several strains expressing GCaMP8f (fast variant of the indicator). Utilizing the myo-2 promoter, we compare pharyngeal muscle activity measured with GCaMP7f and GCaMP8f and find that GCaMP8f is brighter upon binding to calcium, shows faster kinetics, and is not disruptive to the intrinsic contraction dynamics of the pharynx. Additionally, we validate its application for detecting neuronal activity in touch receptor neurons which reveals robust calcium transients even at small stimulus amplitudes. As such, we establish GCaMP8f as a potent tool for C. elegans research which is capable of extracting fast calcium dynamics at very low magnifications across multiple cell types.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11457936PMC
http://dx.doi.org/10.1093/genetics/iyae125DOI Listing

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