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Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons
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Correlative Live-Cell and Super-Resolution Imaging to Link Presynaptic Molecular Organisation With Function
Rachel E Jackson1,2, Benjamin Compans1,2, Juan Burrone1,2
1Centre for Developmental Neurobiology, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United Kingdom.
Frontiers in Synaptic Neuroscience
|March 4, 2022
Summary
Researchers developed a new imaging technique to link synaptic protein organization to neurotransmitter release. This method reveals how nanoscale structures at synapses influence communication efficiency.
Area of Science:
- Neuroscience
- Cell Biology
- Microscopy
Background:
- Synaptic transmission relies on vesicle fusion and neurotransmitter release, but the origins of variability are unclear.
- Super-resolution microscopy shows precise synaptic protein organization, influencing neurotransmission.
- The link between nanoscale protein organization and release probability remains poorly understood.
Purpose of the Study:
- To investigate the structure-function relationship at individual presynaptic boutons.
- To determine if nanoscale organization of release machinery proteins affects neurotransmitter release probability.
- To develop a pipeline for correlative functional and super-resolution microscopy.
Main Methods:
- Developed a pipeline combining live imaging with post-hoc immunolabelling.
- Utilized SypHy-RGECO, a dual reporter for calcium influx and neurotransmitter release.
- Employed multicolour single-molecule localization microscopy (SMLM).
Main Results:
- Demonstrated a novel correlative imaging approach for synaptic studies.
- Enabled simultaneous measurement of presynaptic calcium and neurotransmitter release.
- Provided a framework to link nanoscale organization to synaptic function.
Conclusions:
- The developed pipeline facilitates investigation of structure-function relationships at individual synapses.
- This approach can elucidate how nanoscale protein organization impacts neurotransmitter release variability.
- Advances in multicolour imaging enable detailed analysis of synaptic protein nanostructure.
