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Designer indicators for two-photon recording of subthreshold voltage dynamics
Michelle Ann Land1, Mario Galdamez1, Vincent Villette2
1Department of Neuroscience, Baylor College of Medicine, Houston, TX, USA.
Biorxiv : the Preprint Server for Biology
|April 17, 2026
Summary
New genetically encoded voltage indicators (GEVIs) called JEDI3sub and JEDI3hyp enable sensitive in vivo optical recording of neuronal subthreshold voltage dynamics using two-photon microscopy.
Area of Science:
- Neuroscience
- Molecular Biology
- Biophysics
Background:
- Subthreshold voltage dynamics are crucial for neuronal function but difficult to study in vivo.
- Existing genetically encoded voltage indicators (GEVIs) lack the sensitivity for detecting millivolt-scale subthreshold signals with two-photon (2P) microscopy.
Purpose of the Study:
- To develop novel GEVIs with enhanced sensitivity for subthreshold voltage detection in vivo.
- To overcome limitations of current tools for deep-tissue neuronal recording.
Main Methods:
- Refinement of a multiparametric two-photon high-throughput screening platform.
- Development and testing of two new GEVIs: JEDI3sub and JEDI3hyp.
- Fast 2P optical recording in awake, behaving mice.
Main Results:
- JEDI3 indicators demonstrated superior sensitivity for subthreshold voltage detection compared to JEDI-2P.
- JEDI3sub tracked population-level subthreshold optical tuning.
- JEDI3hyp captured subthreshold dynamics in hippocampal interneurons and enabled deep-tissue imaging of voltage changes.
Conclusions:
- JEDI3sub and JEDI3hyp significantly advance the capability for 2P optical recording of subthreshold voltage dynamics in vivo.
- These novel GEVIs open new research avenues for understanding neural information processing and its role in neurological health and disease.
