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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Abstract 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.
AbstractList 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.
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.
Author Wood, Courtney P.
Wiley, Mark B.
Perez, Pedro A.
Argueta, Donovan A.
Avalos, Bryant
DiPatrizio, Nicholas V.
AuthorAffiliation 1 Division of Biomedical Sciences, School of Medicine, University of California, Riverside , Riverside, CA , United States
2 Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of California, Irvine , Irvine, CA , United States
AuthorAffiliation_xml – name: 2 Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of California, Irvine , Irvine, CA , United States
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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
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Snippet The endocannabinoid system is expressed in cells throughout the body and controls a variety of physiological and pathophysiological functions. We describe...
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StartPage 699712
SubjectTerms endocannabinoids
enzyme kinetics
fatty acid amide hydrolase
lipid metabolism
monoacylglycerol lipase
Physiology
UPLC-MS/MS
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Title UPLC-MS/MS Method for Analysis of Endocannabinoid and Related Lipid Metabolism in Mouse Mucosal Tissue
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Volume 12
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