title: Characterization of the endocannabinoid system in mouse embryonic stem cells.
abstract: In this study, we have ascertained the presence and functionality in mouse embryonic stem cells (ESCs) of members of the endocannabinoid system that have been proposed as possible modulators of the survival and differentiation of various type of stem cells. We show that mouse ESCs, in addition to classical CB(1) and CB(2) cannabinoid receptors, express the transient receptor potential vanilloid receptor, at mRNA, protein, and binding levels. Remarkably, we demonstrate that ESCs have the mRNA, protein, and enzyme activity to synthesize and degrade the prominent endocannabinoids anandamide (through N-acyl-phosphatidylethanolamine-specific phospholipase D and fatty acid amide hydrolase) and 2-arachidonoylglycerol (through diacylglycerol lipase and monoacylglycerol lipase). In addition, both endocannabinoids were detected in ESCs that were also shown to constitutively release a fatty acid amide hydrolase-activating compound. Finally, we document that the stimulation of ESCs by methanandamide, a nonhydrolysable analog of anandamide, does not lead to overt alteration of the expression of Oct3/4, Nanog, and Cdx2, genes that are involved in early cell fate in the preimplantation embryo and stemness, or of the expression patterns of Brachyury and Hnf4, genes that are used as late markers of lineage differentiation capability of ESC-derived embryoid bodies. Similarly ineffective on the expression of the tested stemness genes was 2-arachidonoylglycerol. Taken together, these results confirm and extend the notion that ESCs express several functional members of the endocannabinoid system, but they leave open the question about their role in stem cells as modulators of stemness and differentiation potential.
authors.affiliation: Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Bari
authors.forename: Marianna
authors.initials: M
authors.lastname: Tedesco
authors.forename: Natalia
authors.initials: N
authors.lastname: Battista
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Pasquariello
authors.forename: Mariangela
authors.initials: M
authors.lastname: Pucci
authors.forename: Valeria
authors.initials: V
authors.lastname: Gasperi
authors.forename: Maria Lucia
authors.initials: ML
authors.lastname: Scaldaferri
authors.forename: Donatella
authors.initials: D
authors.lastname: Farini
authors.forename: Massimo
authors.initials: M
authors.lastname: De Felici
authors.forename: Mauro
authors.initials: M
authors.lastname: Maccarrone
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D063385:Cannabinoid Receptor Modulators / Q000235:genetics / Q000378:metabolism*; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D017077:Culture Media, Conditioned / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D063388:Endocannabinoids; D020536:Enzyme Activators / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D005989:Glycerides / Q000494:pharmacology; D051379:Mice; D053284:Polyunsaturated Alkamides / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D001095:Arachidonic Acids; D015415:Biomarkers; D063385:Cannabinoid Receptor Modulators; D017077:Culture Media, Conditioned; D063388:Endocannabinoids; D020536:Enzyme Activators; D005989:Glycerides; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger; D000099252:Fatty Acid Amide Hydrolases; C088155:methanandamide; C094503:glyceryl 2-arachidonate; C078814:anandamide