CB₂ cannabinoid receptors inhibit synaptic transmission when expressed in cultured autaptic neurons

Neuropharmacology. 2012 Sep;63(4):514-23. doi: 10.1016/j.neuropharm.2012.04.024. Epub 2012 May 8.

Abstract

The role of CB₂ in the central nervous system, particularly in neurons, has generated much controversy. Fueling the controversy are imperfect tools, which have made conclusive identification of CB₂ expressing neurons problematic. Imprecise localization of CB₂ has made it difficult to determine its function in neurons. Here we avoid the localization controversy and directly address the question if CB₂ can modulate neurotransmission. CB₂ was expressed in excitatory hippocampal autaptic neurons obtained from CB₁ null mice. Whole-cell patch clamp recordings were made from these neurons to determine the effects of CB₂ on short-term synaptic plasticity. CB₂ expression restored depolarization induced suppression of excitation to these neurons, which was lost following genetic ablation of CB₁. The endocannabinoid 2-arachidonylglycerol (2-AG) mimicked the effects of depolarization in CB₂ expressing neurons. Interestingly, ongoing basal production of 2-AG resulted in constitutive activation of CB₂, causing a tonic inhibition of neurotransmission that was relieved by the CB₂ antagonist AM630 or the diacylglycerol lipase inhibitor RHC80267. Through immunocytochemistry and analysis of spontaneous EPSCs, paired pulse ratios and coefficients of variation we determined that CB₂ exerts its function at a presynaptic site of action, likely through inhibition of voltage gated calcium channels. Therefore CB₂ expressed in neurons effectively mimics the actions of CB₁. Thus neuronal CB₂ is well suited to integrate into conventional neuronal endocannabinoid signaling processes, with its specific role determined by its unique and highly inducible expression profile.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Arachidonic Acids / antagonists & inhibitors
  • Arachidonic Acids / metabolism
  • Arachidonic Acids / pharmacology
  • Astrocytes / cytology
  • Cells, Cultured
  • Endocannabinoids / antagonists & inhibitors
  • Endocannabinoids / metabolism
  • Endocannabinoids / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Glycerides / antagonists & inhibitors
  • Glycerides / metabolism
  • Glycerides / pharmacology
  • Heterozygote
  • Hippocampus / cytology
  • Hippocampus / drug effects
  • Hippocampus / metabolism*
  • Lipoprotein Lipase / antagonists & inhibitors
  • Mice
  • Mice, Knockout
  • Nerve Tissue Proteins / agonists
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism*
  • Patch-Clamp Techniques
  • Presynaptic Terminals / drug effects
  • Presynaptic Terminals / metabolism
  • Receptor, Cannabinoid, CB1 / genetics
  • Receptor, Cannabinoid, CB1 / metabolism
  • Receptor, Cannabinoid, CB2 / agonists
  • Receptor, Cannabinoid, CB2 / antagonists & inhibitors
  • Receptor, Cannabinoid, CB2 / genetics
  • Receptor, Cannabinoid, CB2 / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Synaptic Transmission* / drug effects

Substances

  • Arachidonic Acids
  • Cnr2 protein, mouse
  • Endocannabinoids
  • Enzyme Inhibitors
  • Glycerides
  • Nerve Tissue Proteins
  • Receptor, Cannabinoid, CB1
  • Receptor, Cannabinoid, CB2
  • Recombinant Fusion Proteins
  • glyceryl 2-arachidonate
  • Lipoprotein Lipase