Subcellular patch-clamp techniques for single-bouton stimulation and simultaneous pre- and postsynaptic recording at
David Vandael1, Yuji Okamoto1, Carolina Borges-Merjane1
1IST Austria (Institute of Science and Technology Austria), Klosterneuburg, Austria.
Nature Protocols
|May 15, 2021
Summary
This study presents a new subcellular patch-clamp method for recording from small cortical synapses in brain slices. This technique allows detailed analysis of synaptic transmission and neuronal circuit function.
Area of Science:
- Neuroscience
- Cell Biology
Background:
- Simultaneous recording from presynaptic terminals and postsynaptic neurons is crucial for studying synaptic transmission.
- Small cortical presynaptic terminals have been difficult to access for direct recording.
Purpose of the Study:
- To develop and validate an improved subcellular patch-clamp recording protocol for small mammalian cortical synapses.
- To enable detailed analysis of synaptic structure and function in an intact brain slice environment.
Main Methods:
- Developed a subcellular patch-clamp technique for recording from single presynaptic hippocampal mossy fiber terminals in rat and mouse brain slices.
- Utilized bouton-attached and whole-bouton configurations for recording.
- Integrated recordings with biocytin labeling, morphological analysis, axon tract stimulation, optogenetics, and spontaneous event analysis.
Main Results:
- Successfully obtained paired pre- and postsynaptic recordings from hippocampal mossy fiber terminals.
- Demonstrated the applicability of the technique to genetically modified models and combined it with various stimulation and analysis methods.
- Enabled correlative investigation of synapse structure and function, and analysis of miniature events at single synapses.
Conclusions:
- The described subcellular patch-clamp techniques significantly advance the ability to analyze the biophysics, plasticity, and circuit function of mammalian cortical synapses.
- This protocol provides a powerful tool for neuroscience research, particularly for studying genetically modified models and complex synaptic events.
