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

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.