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The serine hydrolase ABHD6 controls the accumulation and efficacy of 2-AG at cannabinoid receptors

Nature Neuroscience volume 13, pages 951–957 (2010)Cite this article

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Abstract

The endocannabinoid 2-arachidonoylglycerol (2-AG) regulates neurotransmission and neuroinflammation by activating CB1 cannabinoid receptors on neurons and CB2 cannabinoid receptors on microglia. Enzymes that hydrolyze 2-AG, such as monoacylglycerol lipase, regulate the accumulation and efficacy of 2-AG at cannabinoid receptors. We found that the recently described serine hydrolase α-β-hydrolase domain 6 (ABHD6) also controls the accumulation and efficacy of 2-AG at cannabinoid receptors. In cells from the BV-2 microglia cell line, ABHD6 knockdown reduced hydrolysis of 2-AG and increased the efficacy with which 2-AG can stimulate CB2-mediated cell migration. ABHD6 was expressed by neurons in primary culture and its inhibition led to activity-dependent accumulation of 2-AG. In adult mouse cortex, ABHD6 was located postsynaptically and its selective inhibition allowed the induction of CB1-dependent long-term depression by otherwise subthreshold stimulation. Our results indicate that ABHD6 is a rate-limiting step of 2-AG signaling and is therefore a bona fide member of the endocannabinoid signaling system.

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Figure 1: ABHD6 hydrolyzes 2-AG in BV-2 cells and controls the efficacy of 2-AG at CB2 receptors.
Figure 2: Effect of the ABHD6 inhibitor WWL70 and the MAGL inhibitor JZL184 on [3H]–2-AG hydrolysis in cell homogenates.
Figure 3: Effect of WWL70 and JZL184 on [3H]–2-AG hydrolysis and 2-AG accumulation in intact neurons in primary culture.
Figure 4: Visualization of ABHD6 protein in different cell types.
Figure 5: Localization of ABHD6 protein in mouse prefrontal cortex.
Figure 6: Effect of WWL70 and JZL184 on CB1-dependent LTD in mouse prefrontal cortex.

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Acknowledgements

This work was supported by grants from the National Institute on Drug Abuse (DA14486 and DA26430 to N.S., DA017259, DA009789 and DA025285 to B.F.C. and DA026161 to J.L.B.) and from the National Institute of General Medical Sciences (PHS NRSA 2T32 GM007270 to W.R.M.).

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Author notes

  1. Jacqueline L Blankman and Eric A Horne: These authors contributed equally to this work.

Authors and Affiliations

  1. Neurobiology and Behavior Graduate Program, University of Washington, Seattle, Washington, USA

    William R Marrs & Susan Fung

  2. The Skaggs Institute for Chemical Biology and Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA

    Jacqueline L Blankman, Jessica P Alexander, Jonathan Z Long, Weiwei Li & Benjamin F Cravatt

  3. Department of Pharmacology, University of Washington, Seattle, Washington, USA

    Eric A Horne, Yi Hsing Lin, Jonathan Coy, Cong Xu & Nephi Stella

  4. INSERM U862, Equipe Physiopathologie de la plasticité synaptique, Bordeaux, France

    Aurore Thomazeau, Mathieu Lafourcade & Olivier J Manzoni

  5. Department of Otolaryngology, University of Washington, Seattle, Washington, USA

    Agnes L Bodor

  6. Bioanalysis and Pharmacology of Bioactive Lipids Laboratory, CHAM7230, Louvain Drug Research Institute, Université catholique de Louvain, Bruxelles, Belgium

    Giulio G Muccioli

  7. Department of Psychological and Brain Sciences, Gill Center for Biomolecular Science, Indiana University, Bloomington, Indiana, USA

    Sherry Shu-Jung Hu & Ken Mackie

  8. Neurobiology Undergraduate Program, University of Washington, Seattle, Washington, USA

    Grace Woodruff

  9. Department of Neurology, University of Washington, Seattle, Washington, USA

    Thomas Möller

  10. Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, Washington, USA

    Nephi Stella

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Contributions

W.R.M. prepared the cell cultures, performed the hydrolysis experiments, conducted the data analysis and wrote the manuscript. J.L.B. performed the ABPP experiments and contributed to the data analysis. E.A.H. performed the GC-MS and immunofluoresence experiments and contributed to the electron microscopy experiments and data analysis. A.T. and M.L. performed the electrophysiology experiments. Y.H.L. prepared the cell culture transfections and the shRNA constructs, and performed the qPCR experiments. J.C. contributed to the immunofluorescence experiments. A.L.B. performed the electron microscopy experiments. G.G.M. contributed to the hydrolysis experiments. S.S.-J.H. contributed to antibody production. G.W. and S.F. performed the cell migration experiments. J.P.A. contributed to the ABPP experiments. J.Z.L. and W.L. produced the hydrolase inhibitors. C.X. contributed to the cell culture experiments. T.M. provided the transgenic mice. K.M. provided antibodies. O.J.M. supervised the electrophysiology experiments. B.F.C. supervised the ABPP experiments and the development of hydrolase inhibitors. N.S. supervised the project and wrote the manuscript.

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Correspondence to Nephi Stella.

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Marrs, W., Blankman, J., Horne, E. et al. The serine hydrolase ABHD6 controls the accumulation and efficacy of 2-AG at cannabinoid receptors. Nat Neurosci 13, 951–957 (2010). https://doi.org/10.1038/nn.2601

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