Assay of Endocannabinoid Uptake

Ines Reynoso-Moreno; Mark Rau; Andrea Chicca; Simon Nicolussi; Jürg Gertsch

doi:10.1007/978-1-0716-2728-0_28

Assay of Endocannabinoid Uptake

Methods Mol Biol. 2023:2576:329-348. doi: 10.1007/978-1-0716-2728-0_28.

Authors

Ines Reynoso-Moreno¹, Mark Rau¹, Andrea Chicca¹, Simon Nicolussi¹, Jürg Gertsch²

Affiliations

¹ Institute of Biochemistry and Molecular Medicine, NCCR TransCure, University of Bern, Bern, Switzerland.
² Institute of Biochemistry and Molecular Medicine, NCCR TransCure, University of Bern, Bern, Switzerland. gertsch@ibmm.unibe.ch.

PMID: 36152200
DOI: 10.1007/978-1-0716-2728-0_28

Abstract

Endocannabinoids at nanomolar physiological concentrations cross cellular membranes by facilitated diffusion, a process that can be studied by measuring transport kinetics and endocannabinoid trafficking employing radioligands and mass spectrometry. Here, we describe radiosubstrate-based assays using arachidonoyl[1-³H]ethanolamine and 2-arachidonoyl[1,2,3-³H]glycerol to measure cellular endocannabinoid uptake in a three-phase assay with human U937 cells. Liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS)-based lipidomics was used to interrogate the roles of serum and albumin for endocannabinoid trafficking in U937 cells.

Keywords: 2-Arachidonoylglycerol; Anandamide; Endocannabinoid uptake; LC–ESI–MS/MS.

MeSH terms

Albumins
Endocannabinoids*
Ethanolamines
Glycerol
Humans
Tandem Mass Spectrometry* / methods

Substances

Albumins
Endocannabinoids
Ethanolamines
Glycerol