Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2576:329-348.
doi: 10.1007/978-1-0716-2728-0_28.

Assay of Endocannabinoid Uptake

Ines Reynoso-Moreno  1 , Mark Rau  1 , Andrea Chicca  1 , Simon Nicolussi  1 , Jürg Gertsch  2
Affiliations

Assay of Endocannabinoid Uptake

Ines Reynoso-Moreno et al. Methods Mol Biol. 2023.

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.

PubMed Disclaimer

References

    1. Nicolussi S, Gertsch JJ (2015) Endocannabinoid transport revisited. Vitam Horm 98:441–485 - DOI
    1. Fowler CJ, Tiger G, Ligresti A et al (2004) Selective inhibition of anandamide cellular uptake versus enzymatic hydrolysis—a difficult issue to handle. Eur J Pharmacol 492:1–11 - DOI
    1. Oddi S, Fezza F, Catanzaro G et al (2010) Pitfalls and solutions in assaying anandamide transport in cells. J Lipid Res 51:2435–2444 - DOI
    1. Nicolussi S, Chicca A, Rau M et al (2014) Correlating FAAH and anandamide cellular uptake inhibition using N-alkylcarbamate inhibitors: From ultrapotent to hyperpotent. Biochem Pharmacol 92:669–689 - DOI
    1. Fowler CJ (2013) Transport of endocannabinoids across the plasma membrane and within the cell. FEBS J 280:1895–1904 - DOI

LinkOut - more resources