Abstract
The endocannabinoids, 2-arachidonoylglycerol (2-AG) and arachidonylethanolamide (AEA), are endogenous ligands for the cannabinoid receptors (CB1 and CB2) and are implicated in a wide array of physiological processes. These neutral arachidonic acid (AA) derivatives have been identified as efficient substrates for the second isoform of the cyclooxygenase enzyme (COX-2). A diverse family of prostaglandin glycerol esters (PG-Gs) and prostaglandin ethanolamides (PG-EAs) is generated by the action of COX-2 (and downstream prostaglandin synthases) on 2-AG and AEA. As the biological importance of the endocannabinoid system becomes more apparent, there is a tremendous need for robust, sensitive, and efficient analytical methodology for the endocannabinoids and their metabolites. In this chapter, we describe methodology suitable for carrying out oxygenation of endocannabinoids by COX-2, and analysis of products of endocannabinoid oxygenation by COX-2 and of endocannabinoids themselves from in vitro and cell assays.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Devane WA, Hanus L, Breuer A et al (1992) Isolation and structure of a brain constituent that binds to the cannabinoid receptor. Science 258:1946–1949
Stella N, Schweitzer P, Piomelli D (1997) A second endogenous cannabinoid that modulates long-term potentiation. Nature 388:773–778
Kozak KR, Rowlinson SW, Marnett LJ (2000) Oxygenation of the endocannabinoid, 2-arachidonylglycerol, to glyceryl prostaglandins by cyclooxygenase-2. J Biol Chem 275:33744–33749
Yu M, Ives D, Ramesha CS (1997) Synthesis of prostaglandin E2 ethanolamide from anandamide by cyclooxygenase-2. J Biol Chem 272:21181–21186
Sang N, Zhang J, Chen C (2006) PGE2 glycerol ester, a COX-2 oxidative metabolite of 2-arachidonoyl glycerol, modulates inhibitory synaptic transmission in mouse hippocampal neurons. J Physiol (Lond) 572:735–745
Sang N, Zhang J, Chen C (2007) COX-2 oxidative metabolite of endocannabinoid 2-AG enhances excitatory glutamatergic synaptic transmission and induces neurotoxicity. J Neurochem 102:1966–1977
Nirodi CS, Crews BC, Kozak KR et al (2004) The glyceryl ester of prostaglandin E2 mobilizes calcium and activates signal transduction in RAW264.7 cells. Proc Natl Acad Sci U S A 101:1840–1845
Hu SS-J, Bradshaw HB, Chen JSC et al (2008) Prostaglandin E-2 glycerol ester, an endogenous COX-2 metabolite of 2-arachidonoylglycerol, induces hyperalgesia and modulates nf kappa b activity. Br J Pharmacol 153:1538–1549
Valdeolivas S, Pazos MR, Bisogno T et al (2013) The inhibition of 2-arachidonoyl-glycerol (2-AG) biosynthesis, rather than enhancing striatal damage, protects striatal neurons from malonate-induced death: A potential role of cyclooxygenase-2-dependent metabolism of 2-AG. Cell Death Dis 4:e862
Alhouayek M, Masquelier J, Cani PD et al (2013) Implication of the anti-inflammatory bioactive lipid prostaglandin d2-glycerol ester in the control of macrophage activation and inflammation by abhd6. Proc Natl Acad Sci U S A 110:17558–17563
Rouzer CA, Marnett LJ (2005) Glycerylprostaglandin synthesis by resident peritoneal macrophages in response to a zymosan stimulus. J Biol Chem 280:26690–26700
Kingsley PJ, Marnett LJ (2003) Analysis of endocannabinoids by Ag+ coordination tandem mass spectrometry. Anal Biochem 314:8–15
Rouzer CA, Ghebreselasie K, Marnett LJ (2002) Chemical stability of 2-arachidonylglycerol under biological conditions. Chem Phys Lipids 119:69–82
Kingsley PJ, Rouzer CA, Saleh S et al (2005) Simultaneous analysis of prostaglandin glyceryl esters and prostaglandins by electrospray tandem mass spectrometry. Anal Biochem 343:203–211
Weber A, Ni JS, Ling KHJ et al (2004) Formation of prostamides from anandamide in FAAH knockout mice analyzed by HPLC with tandem mass spectrometry. J Lipid Res 45:757–763
Marnett LJ, Rowlinson SW, Goodwin DC et al (1999) Arachidonic acid oxygenation by COX-1 and COX-2 - mechanisms of catalysis and inhibition. J Biol Chem 274:22903–22906
Acknowledgements
We are grateful to Carol Rouzer for editorial assistance. This work was supported by a research grant from the National Institutes of Health (GM 15431) to L.J.M.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kudalkar, S.N., Kingsley, P.J., Marnett, L.J. (2016). Assay of Endocannabinoid Oxidation by Cyclooxygenase-2. In: Maccarrone, M. (eds) Endocannabinoid Signaling. Methods in Molecular Biology, vol 1412. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3539-0_21
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3539-0_21
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3537-6
Online ISBN: 978-1-4939-3539-0
eBook Packages: Springer Protocols