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Functional Analysis of Mitochondrial CB1 Cannabinoid Receptors (mtCB1) in the Brain.

By July 14, 2017No Comments
Methods Enzymol. 2017;593:143-174. doi: 10.1016/bs.mie.2017.06.023. Epub 2017 Jul 14.

Abstract

PM 2 site 207Recent evidence indicates that, besides its canonical localization at cell plasma membranes, the type-1 cannabinoid receptor, CB1 is functionally present at brain and muscle mitochondrial membranes (mtCB1). Through mtCB1 receptors, cannabinoids can directly regulate intramitochondrial signaling and respiration. This new and surprising discovery paves the way to new potential fields of research, dealing with the direct impact of G protein-coupled receptors on bioenergetic processes and its functional implications. In this chapter, we summarize some key experimental approaches established in our laboratories to identify anatomical, biochemical, and functional features of mtCB1 receptors in the brain. In particular, we describe the procedures to obtain reliable and controlled detection of mtCB1 receptors by immunogold electromicroscopy and by immunoblotting methods. Then, we address the study of direct cannabinoid effects on the electron transport system and oxidative phosphorylation. Finally, we present a functional example of the impact of mtCB1 receptors on mitochondrial mobility in cultured neurons. Considering the youth of the field, these methodological approaches will very likely be improved and refined in the future, but this chapter aims at presenting the methods that are currently used and, in particular, at underlining the need of rigorous controls to obtain reliable results. We hope that this chapter might help scientists becoming interested in this new and exciting field of research.

KEYWORDS:

Brain; Cannabinoid receptor CB1; Cannabinoids; Complex I activity; Electron microscopy; Immunoprecipitation; Mitochondria; Mitochondrial mobility; OXPHOS; Western immunoblotting

PMID: 28750801

 

DOI: 10.1016/bs.mie.2017.06.023
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