Multiple forms of endocannabinoid and endovanilloid signaling regulate the tonic control of GABA release

PersistentCB₁cannabinoidreceptoractivitylimitsneurotransmitterreleaseatvarioussynapsesthroughoutthebrain.However,itisnotfullyunderstood how constitutively active CB₁ receptors, tonic endocannabinoid signaling, and its regulation by multiple serine hydrolases contribute to the synapsespecific calibration of neurotransmitter release probability. To address this question at perisomatic and dendritic GABAergic synapses in the mouse hippocampus,weusedacombinationof paired whole-cellpatch-clamprecording, liquidchromatography/tandemmassspectrometry, stochastic optical reconstruction microscopy super-resolution imaging, and immunogold electron microscopy. Unexpectedly, application of the CB1 antagonist and inverseagonistAM251[N-1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-1-piperidinyl-1H-pyrazole-3-carboxamide],butnottheneutralantagonist NESS0327 [8-chloro-1-(2,4-dichlorophenyl)-N-piperidin-1-yl-5,6-dihydro-4H-benzo[2,3]cyclohepta[2,4-b]pyrazole-3-carboxamine], significantly increased synaptic transmission between CB1-positive perisomatic interneurons and CA1 pyramidal neurons. JZL184 (4-nitrophenyl 4-[bis(1,3-benzodioxol-5-yl)(hydroxy)methyl]piperidine-1-carboxylate), a selective inhibitor of monoacylglycerol lipase (MGL), the presynaptic degrading enzyme of the endocannabinoid 2-arachidonoylglycerol (2-AG), elicited a robust increase in 2-AG levels and concomitantly decreased GABAergic transmission. In contrast, inhibition of fatty acid amide hydrolase (FAAH) by PF3845 (N-pyridin-3-yl-4-[[3-[5- (trifluoromethyl)pyridin-2-yl]oxyphenyl]methyl]piperidine-1-carboxamide) elevated endocannabinoid/endovanilloid anandamide levels but did not change GABAergic synaptic activity. However, FAAH inhibitors attenuated tonic 2-AG increase and also decreased its synaptic effects. This antagonistic interaction required the activation of the transient receptor potential vanilloid receptor TRPV1, which was concentrated on postsynaptic intracellular membrane cisternae at perisomatic GABAergic symmetrical synapses. Interestingly, neither AM251, JZL184, nor PF3845 affected CB₁-positive dendritic interneuron synapses. Together, these findings are consistent with the possibility that constitutively activeCB₁ receptors substantially influence perisomaticGABArelease probability and indicate that the synaptic effects of tonic 2-AG release are tightly controlled by presynapticMGLactivity and also by postsynaptic endovanilloid signaling and FAAH activity.