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Preparation of Synaptoneurosomes from Mouse Cortex using a Discontinuous Percoll-Sucrose Density Gradient
Published on: September 17, 2011
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Synaptoneurosome Preparation from C57BL/6 Striata
Emma Puighermanal1,2,3, Anne Biever1,2,3, Emmanuel Valjent1,2,3
1CNRS, UMR5203, Institut de Génomique Fonctionnelle, Montpellier, France.
Bio-Protocol
|January 20, 2025
Summary
Synaptic plasticity relies on local mRNA translation, regulated by protein phosphorylation. This study details a method to analyze synaptic phospho-proteins in mouse striatal synaptoneurosomes, aiding research into synaptic remodeling.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Activity-dependent local mRNA translation is crucial for synaptic plasticity, enabling structural and functional remodeling.
- Protein phosphorylation of translational machinery components, including initiation factors and ribosomal proteins, tightly controls this process.
Purpose of the Study:
- To describe a reproducible method for preparing striatal synaptoneurosomes from adult mice.
- To enable quantification of synaptic phospho-proteins within these isolated synaptoneurosomes.
Main Methods:
- Preparation of striatal synaptoneurosomes from adult mice.
- Isolation of synaptoneurosomes containing both pre- and postsynaptic elements.
- Quantification of synaptic phospho-protein levels.
Main Results:
- The protocol successfully yields synaptoneurosomes suitable for phospho-protein analysis.
- The method allows for the assessment of phosphorylation states within the synaptic compartment.
Conclusions:
- This protocol provides a valuable tool for investigating the role of phosphorylation in activity-dependent mRNA translation at synapses.
- Understanding synaptic phospho-protein regulation is key to elucidating mechanisms of synaptic plasticity and function.
