A modular system to label endogenous presynaptic proteins using split fluorophores in Caenorhabditis elegans

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• Visualizing presynaptic protein localization is crucial for understanding synapse development.
• Existing methods may have limitations in live-animal studies.

Purpose of the Study:

• To develop a versatile system for visualizing endogenous presynaptic protein localization in live Caenorhabditis elegans.
• To enable single-neuron resolution studies of synapse formation and patterning.

Main Methods:

• Utilized CRISPR/Cas9 genome editing to generate C. elegans strains.
• Tagged endogenous presynaptic proteins (RAB-3, SNG-1, CLA-1, SYD-2, UNC-10, RIMB-1, ELKS-1) with split fluorescent proteins (GFP11 and wrmScarlet11).
• Expressed split fluorescent protein components (GFP1-10, wrmScarlet1-10) under neuron-specific promoters.

Main Results:

• Successfully visualized endogenous presynaptic proteins at single-neuron resolution in C. elegans.
• Demonstrated robust labeling of presynaptic proteins in various neuron types.
• Created a collection of knock-in strains and complementary plasmids.

Conclusions:

• The developed system provides a modular and versatile tool for neuroscientists.
• Enables detailed examination of endogenous presynaptic protein localization in any C. elegans neuron type.
• Facilitates research into the genetic and molecular mechanisms of synapse formation.