ABHD6 selectively controls metabotropic-dependent increases in 2-AG production

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract The most abundant endocannabinoid ( eCB ) in the brain, 2-arachidonoyl glycerol ( 2-AG) , is hydrolyzed by α/β-hydrolase domain containing 6 ( ABHD6 ); yet how ABHD6 controls stimuli-dependent increases in 2-AG production is unknown. To explore this question, we leveraged the recently developed 2-AG sensor, GRAB eCB2.0 , and found that stimulation of Neuro2a cells in culture with bradykinin ( BK ) acting at metabotropic B 2 K receptors and ATP acting at ionotropic P2X 7 receptors led to differential increases in 2-AG levels. B 2 K triggered increases in 2-AG levels via diacylglycerol lipase ( DAGL ), and this mechanism was potentiated by increases in intracellular calcium and ABHD6 inhibition. By contrast, P2X 7 -triggered increases in 2-AG levels were dependent on DAGL and extracellular calcium but unaffected by ABHD6 inhibition. Thus, ABHD6 preferentially regulates metabotropic-dependent increases in 2-AG levels over ionotropic-dependent increases in 2-AG levels. Our study indicates that ABHD6 selectively controls stimuli-dependent increases in 2-AG production and emphasizes its specific role in eCB signaling.

Authors 11

1. Simar J. Singh

   University of Washington

   Affiliation as printed

   Departments of Pharmacology, University of Washington, Seattle, USA

2. Dennis Sarroza

   University of Washington

   Affiliation as printed

   Departments of Pharmacology, University of Washington, Seattle, USA

3. Anthony English

   University of Washington

   Affiliation as printed

   Departments of Pharmacology, University of Washington, Seattle, USA

4. Dale Whittington

   University of Washington

   Affiliation as printed

   Medicinal Chemistry, University of Washington, Seattle, USA

5. Ao Dong

   Peking University · Center for Life Sciences

   Affiliation as printed

   Peking University School of Life Sciences, PKU-IDG/McGovern Institute for Brain Research, Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China

6. Mario van der Stelt Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands

7. Yulong Li

   Peking University · Center for Life Sciences

   Affiliation as printed

   Peking University School of Life Sciences, PKU-IDG/McGovern Institute for Brain Research, Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China

8. Larry S. Zweifel

   University of Washington

   Affiliation as printed

   Departments of Center for Cannabis Research, University of Washington, Seattle, USA

   Departments of Center for the Neurobiology of Addiction, Pain, and Emotion, University of Washington, Seattle, USA

   Departments of Psychiatry and Behavioral Sciences, University of Washington, Seattle, USA

9. Michael R. Bruchas

   University of Washington

   Affiliation as printed

   Departments of Anesthesiology and Pain Medicine, University of Washington, Seattle, USA

   Departments of Center for Cannabis Research, University of Washington, Seattle, USA

   Departments of Center for the Neurobiology of Addiction, Pain, and Emotion, University of Washington, Seattle, USA

10. Benjamin Bruce Land

    University of Washington

    Affiliation as printed

    Departments of Center for Cannabis Research, University of Washington, Seattle, USA

    Departments of Center for the Neurobiology of Addiction, Pain, and Emotion, University of Washington, Seattle, USA

    Departments of Pharmacology, University of Washington, Seattle, USA

11. Nephi Stella corresponding

    University of Washington

    Affiliation as printed

    Departments of Center for Cannabis Research, University of Washington, Seattle, USA

    Departments of Center for the Neurobiology of Addiction, Pain, and Emotion, University of Washington, Seattle, USA

    Departments of Pharmacology, University of Washington, Seattle, USA

    Departments of Psychiatry and Behavioral Sciences, University of Washington, Seattle, USA