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Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons
Emily R Krueger1, Taylor R Church2, Anna Brennan2
1Department of Molecular Pharmacology and Neuroscience, Loyola University Chicago Stritch School of Medicine, Maywood, IL 60153, USA.
STAR Protocols
|January 11, 2025
Summary
This study outlines a protocol to investigate neuronal membrane proteasomes (NMPs) in mouse sensory neurons. The methods enable analysis of NMP function, expression, and localization, impacting pain sensitivity research.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Neuronal membrane proteasomes (NMPs) are present on specific somatosensory dorsal root ganglion (DRG) neurons.
- NMPs play a role in modulating mechanical sensitivity and pain perception.
Purpose of the Study:
- To provide a detailed protocol for studying the function of NMPs in mouse peripheral sensory neurons.
- To establish methods for analyzing NMP expression, localization, and functional impact.
Main Methods:
- Procurement and primary culture of mouse dorsal root ganglion (DRG) neurons.
- Biochemical assays and antibody feeding techniques for NMP analysis.
- Calcium (Ca2+) imaging to assess NMP function in DRG neurons.
Main Results:
- The protocol successfully details the steps for NMP study in primary DRG neurons.
- Methods for analyzing NMP expression, localization, and function are established.
- Demonstrates the utility of Ca2+ imaging for functional assessment.
Conclusions:
- This protocol facilitates research into the role of NMPs in sensory neuron function.
- Understanding NMP function can provide insights into mechanisms of pain sensitivity.
- The presented methods are valuable for neuroscientists studying peripheral sensory pathways.
