UPLC-MS/MS Method for Analysis of Endocannabinoid and Related Lipid Metabolism in Mouse Mucosal Tissue

The endocannabinoid system is expressed in cells throughout the body and controls a variety of physiological and pathophysiological functions. We describe robust and reproducible UPLC-MS/MS-based methods for analyzing metabolism of the endocannabinoids, 2-arachidonoyl- sn -glycerol and arachidonoyl...

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Published inFrontiers in physiology Vol. 12; p. 699712
Main Authors Wiley, Mark B., Perez, Pedro A., Argueta, Donovan A., Avalos, Bryant, Wood, Courtney P., DiPatrizio, Nicholas V.
Format Journal Article
LanguageEnglish
Published Frontiers Media S.A 14.07.2021
Subjects
endocannabinoids
enzyme kinetics
fatty acid amide hydrolase
lipid metabolism
monoacylglycerol lipase
Physiology
UPLC-MS/MS
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Summary:The endocannabinoid system is expressed in cells throughout the body and controls a variety of physiological and pathophysiological functions. We describe robust and reproducible UPLC-MS/MS-based methods for analyzing metabolism of the endocannabinoids, 2-arachidonoyl- sn -glycerol and arachidonoyl ethanolamide, and related monoacylglycerols (MAGs) and fatty acid ethanolamides (FAEs), respectively, in mouse mucosal tissues (i.e., intestine and lung). These methods are optimized for analysis of activity of the MAG biosynthetic enzyme, diacylglycerol lipase (DGL), and MAG degradative enzymes, monoacylglycerol lipase (MGL) and alpha/beta hydrolase domain containing-6 (ABHD6). Moreover, we describe a novel UPLC-MS/MS-based method for analyzing activity of the FAE degradative enzyme, fatty acid amide hydrolase (FAAH), that does not require use of radioactive substrates. In addition, we describe in vivo pharmacological methods to inhibit MAG biosynthesis selectively in the mouse small-intestinal epithelium. These methods will be useful for profiling endocannabinoid metabolism in rodent mucosal tissues in health and disease.
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Reviewed by: Patrizia Lopalco, University of Bari Aldo Moro, Italy; Giulio G. Muccioli, Catholic University of Louvain, Belgium
This article was submitted to Lipid and Fatty Acid Research, a section of the journal Frontiers in Physiology
Edited by: Simona Lobasso, University of Bari Aldo Moro, Italy
ISSN:1664-042X
1664-042X
DOI:10.3389/fphys.2021.699712