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SynaptoTagMe, a toolkit for in vivo mapping and modulating neurotransmission at single-cell resolution

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Cuentas-Condori, Andrea, Chanabá-López, Patricia, Thomas, Matthew, Feng, Likui, Wolfe, Aaron, Agoba, Peter, Schwartz, Matthew L, Brown, Maximillian, Ebert, Margaret S, Jorgensen, Erik, Bargmann, Cornelia I, Colón-Ramos, Daniel A·2026·eLife
DOI 10.7554/elife.108675PMC PMC13299628

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Abstract

Understanding the organization and regulation of neurotransmission at the level of individual neurons and synapses requires tools that can track and manipulate transmitter-specific vesicles in vivo. Here, we present SynaptoTagMe, a suite of genetic tools in Caenorhabditis elegans to fluorescently label and conditionally ablate the vesicular transporters for glutamate, GABA, acetylcholine, and monoamines. Using a structure-guided approach informed by protein topology and evolutionary conservation, we engineered endogenously tagged versions for each transporter that maintain their physiological function while allowing for cell-specific, bright, and stable visualization. We also developed conditional knockout strains that enable targeted disruption of neurotransmitter synthesis or packaging in single neurons. We applied this toolkit to map co-expression of vesicular transporters across the C. elegans nervous system, revealing that over 10% of neurons exhibit co-transmission. Using the ADF sensory neuron as a case study, we demonstrate that serotonin and acetylcholine are trafficked in partially distinct vesicle pools. Our approach provides a powerful platform for mapping, monitoring, and manipulating neurotransmitter identity and use in vivo. The molecular strategies described here are likely applicable across species, offering a generalizable approach to dissect synaptic communication in vivo.

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Metadata

Authors
Cuentas-Condori, A., Chanabá-López, P., Thomas, M., Feng, L., Wolfe, A., Agoba, P., Schwartz, M. L., Brown, M., Ebert, M. S., Jorgensen, E., Bargmann, C. I., & Colón-Ramos, D. A.
Affiliations
  • Yale University (US)
  • Rockefeller University (US)
  • Howard Hughes Medical Institute (US)
  • Howard Hughes Medical Institute and School of Biological Sciences, University of Utah
  • Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2026-06-25
Volume
14
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.108675
PMC
PMC13299628
OpenAlex
W4415963202
Cited by
0
References
6
Type
posted content
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • National Institutes of Health (R35NS132156, R01NS076558, K99AG083129, F32GM133139, R01NS034307, R01 GM095817)
  • Pew Charitable Trusts (AWD0006561)
  • Jane Coffin Childs Memorial Fund for Medical Research (AWD0006564)
  • Howard Hughes Medical Institute (GT15993)
  • Chan Zuckerberg Initiative
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurobiology and Insect Physiology Research
Keywords
vesicular glutamate transporter, vesicular GABA transporter, vesicular acetylcholine transporter, monoamine transporters, conditional knockout, co-transmission, serotonin, acetylcholine, vesicle pools, Caenorhabditis elegans, single-cell resolution, synaptic transmission
MeSH terms
Animals, Synaptic Transmission, Neurons, Synaptic Vesicles, Caenorhabditis elegans, Neurotransmitter Agents, Synaptotagmins, Single-Cell Analysis

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