Identification of alpha,beta-Hydrolase Domain Containing Protein 6 as a Diacylglycerol Lipase in Neuro-2a Cells

Esbroeck, A.C.M. van; Kantae, V.; Di, X.; Wel, T. van der; Dulk, J. den; Stevens, A.F.; ... ; Stelt, M. van der (2019)

Article / Letter to editor

The endocannabinoid 2-arachidonoylglycerol (2-AG) is involved in neuronal differentiation. This study aimed to identify the biosynthetic enzymes responsible for 2-AG production during retinoic acid (RA)-induced neurite outgrowth of Neuro-2a cells. First, we confirmed that RA stimulation of Neuro-2a cells increases 2-AG production and neurite outgrowth. The diacylglycerol lipase (DAGL) inhibitor DH376 blocked 2-AG production and reduced neuronal differentiation. Surprisingly, CRISPR/Cas9-mediated knockdown of DAGL alpha and DAGL beta in Neuro-2a cells did not reduce 2-AG levels, suggesting another enzyme capable of producing 2-AG in this cell line. Chemical proteomics revealed DAGL beta and alpha,beta-hydrolase domain containing protein (ABHD6) as the only targets of DH376 in Neuro-2a cells. Biochemical, genetic and lipidomic studies demonstrated that ABHD6 possesses DAGL activity in conjunction with its previously reported monoacylglycerol lipase activity. RA treatment of Neuro-2a cells increased by three-fold the amount of active ABHD6. Our study shows that ABHD6 exhibits significant DAG lipase activity in Neuro-2a cells in addition to its known MAG lipase activity and suggest it is involved in neuronal differentiation.

• All authors: Esbroeck, A.C.M. van; Kantae, V.; Di, X.; Wel, T. van der; Dulk, J. den; Stevens, A.F.; Singh, S.; Bakker, A.T.; Florea, B.I.; Stella, N.; Overkleeft, H.S.; Hankemeier, T.; Stelt, M. van der
• Date: 2019-11-26
• Journal: Frontiers in Molecular Neuroscience
• Volume: 12
• Pages: 286
• DOI: doi:10.3389/fnmol.2019.00286