研究小鼠外周感觉神经元中神经膜蛋白酶体功能的实验方案。

Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons.

作者信息

Krueger Emily R, Church Taylor R, Brennan Anna, Türker Fulya, Villalón Landeros Eric

机构信息

Department of Molecular Pharmacology and Neuroscience, Loyola University Chicago Stritch School of Medicine, Maywood, IL 60153, USA.

Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103552. doi: 10.1016/j.xpro.2024.103552. Epub 2025 Jan 10.

DOI:10.1016/j.xpro.2024.103552

PMID:39798095

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11969408/

Abstract

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 Ca 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..

摘要

神经元膜蛋白酶体（NMPs）在一部分躯体感觉背根神经节（DRG）神经元上表达，并影响机械敏感性和疼痛敏感性。在此，我们介绍一种用于研究小鼠外周感觉神经元中NMP功能的方案。我们描述了获取和培养原代DRG神经元的步骤。然后，我们详细介绍了用于分析NMP表达和定位的生化和抗体导入方法。最后，我们纳入了用于研究DRG神经元中NMP功能的钙成像技术。有关本方案使用和执行的完整详细信息，请参阅比利亚隆·兰德罗斯等人的研究。