Depolarizing and calciumamobilizing stimuli fail to enhance synthesis and release of endocannabinoids from rat brain cerebral cortex slices

• Published in: Journal of neurochemistry Vol. 117; no. 4; pp. 665 - 677
• Main Authors: Sarmad, Sarir, Alexander, Stephen PH, Barrett, David A, Marsden, Charles A, Kendall, David A
• Format: Journal Article
• Language: English
• Published: 01.05.2011
• Subjects: Anandamide; Brain slice preparation; Calcium (extracellular); Cannabinoids; Cortex; Depolarization; Esters; Fatty-acid amide hydrolase; Glutamic acid receptors; Ionomycin; Ions; potassium chloride; Supplementation
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/j.1471-4159.2011.07235.x

The concentrations of the endocannabinoids 2-arachidonoylglycerol (2-AG) and N-arachidonylethanolamine (anandamide) were examined in rat brain cerebral cortex slices and surrounding medium. Basal concentrations of endocannabinoids were similar to those identified previously in rat brain, with anandamide content being much lower (19pmol/g) than that of 2-AG (7300pmol/g). In contrast, basal concentrations in the surrounding medium were proportionally much lower for 2-arachidonoylglycerol (16pmol/mL) compared to anandamide (0.6pmol/mL). Incubation of slices with glutamate receptor agonists, depolarizing concentrations of KCl, or ionomycin failed to alter tissue concentrations of endocannabinoids, while endocannabinoids in the medium were unaltered by elevated KCl. Cyclohexyl carbamic acid 3'-carbamoyl-biphenyl-3-yl ester, an inhibitor of fatty acid amide hydrolase, significantly enhanced tissue concentrations of anandamide (and related N-acylethanolamines), without altering 2-AG, while evoking proportional elevations of anandamide in the medium. Removal of extracellular calcium ions failed to alter tissue concentrations of anandamide, but significantly reduced 2-AG in the tissue by 90% and levels in the medium to below the detection limit. Supplementation of the medium with 50 mu M N-oleoylethanolamine only raised tissue concentrations of N-oleoylethanolamine in the presence of cyclohexyl carbamic acid 3'-carbamoyl-biphenyl-3-yl ester and failed to alter either tissue or medium anandamide or 2-AG concentrations. These results highlight the ongoing turnover of endocannabinoids, and the importance of calcium ions in maintaining 2-AG concentrations in this tissue.Original Abstract: J. Neurochem. (2011) 117, 665-677.