Methods to assay anandamide hydrolysis and transport in synaptosomes.
Anandamide (AEA) is the most studied member of a new class of lipid mediators, collectively called endocannabinoids. The biological activity of AEA at cannabinoid and noncannabinoid receptors depends on its life span in the extracellular space, which is regulated by a rapid cellular uptake, followed by intracellular degradation by the enzyme AEA hydrolase (fatty acid amide hydrolase). Here, we present the methodological details of the procedures that we have developed to assay fatty acid amide hydrolase activity and to characterize AEA transport through cell membranes in a new ideal ex vivo system like brain synaptosomes.
Fezza
Filomena
F
Department of Experimental Medicine and Biochemical Sciences, University of Rome, Tor Vergata, Italy.
Battista
Natalia
N
Bari
Monica
M
Maccarrone
Mauro
M
D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D001692:Biological Transport; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D063388:Endocannabinoids; D006868:Hydrolysis; D053284:Polyunsaturated Alkamides; D051381:Rats; D012588:Scintillation Counting; D013574:Synaptosomes / Q000378:metabolism*
D001095:Arachidonic Acids; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; C078814:anandamide