Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons
Summary: Neuronal membrane proteasomes (NMPs) are expressed on a subset of somatosensory dorsal root ganglion (DRG) neurons and influence mechanical and pain sensitivity. Here, we present a protocol for studying NMP function in mouse peripheral sensory neurons. We describe steps for procuring and cu...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007172 |
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| author | Emily R. Krueger Taylor R. Church Anna Brennan Fulya Türker Eric Villalón Landeros |
| author_facet | Emily R. Krueger Taylor R. Church Anna Brennan Fulya Türker Eric Villalón Landeros |
| author_sort | Emily R. Krueger |
| collection | DOAJ |
| description | Summary: Neuronal membrane proteasomes (NMPs) are expressed on a subset of somatosensory dorsal root ganglion (DRG) neurons and influence mechanical and pain sensitivity. Here, we present a protocol for studying NMP function in mouse peripheral sensory neurons. We describe steps for procuring and culturing primary DRG neurons. We then detail biochemical and antibody feeding approaches to analyze NMP expression and localization. Finally, we include Ca2+ imaging techniques for investigating NMP function in DRG neurons.For complete details on the use and execution of this protocol, please refer to Villalón Landeros et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-088a76b7cb0648cc85cd43d39ac06e9f |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-088a76b7cb0648cc85cd43d39ac06e9f2025-01-11T06:41:59ZengElsevierSTAR Protocols2666-16672025-03-0161103552Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neuronsEmily R. Krueger0Taylor R. Church1Anna Brennan2Fulya Türker3Eric Villalón Landeros4Department of Molecular Pharmacology and Neuroscience, Loyola University Chicago Stritch School of Medicine, Maywood, IL 60153, USADepartment of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USADepartment of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USADepartment of Molecular Biology and Genetics, Faculty of Science, İhsan Doğramacı Bilkent University, Ankara 06800, TurkeyDepartment of Molecular Pharmacology and Neuroscience, Loyola University Chicago Stritch School of Medicine, Maywood, IL 60153, USA; Center for Translational Research and Education, Health Sciences Campus, 2160 South First Avenue, Maywood, IL 60153, USA; Corresponding authorSummary: Neuronal membrane proteasomes (NMPs) are expressed on a subset of somatosensory dorsal root ganglion (DRG) neurons and influence mechanical and pain sensitivity. Here, we present a protocol for studying NMP function in mouse peripheral sensory neurons. We describe steps for procuring and culturing primary DRG neurons. We then detail biochemical and antibody feeding approaches to analyze NMP expression and localization. Finally, we include Ca2+ imaging techniques for investigating NMP function in DRG neurons.For complete details on the use and execution of this protocol, please refer to Villalón Landeros et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724007172Cell cultureAntibodyNeuroscienceProtein expression and purification |
| spellingShingle | Emily R. Krueger Taylor R. Church Anna Brennan Fulya Türker Eric Villalón Landeros Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons STAR Protocols Cell culture Antibody Neuroscience Protein expression and purification |
| title | Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons |
| title_full | Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons |
| title_fullStr | Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons |
| title_full_unstemmed | Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons |
| title_short | Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons |
| title_sort | protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons |
| topic | Cell culture Antibody Neuroscience Protein expression and purification |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007172 |
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