Activation of type 5 metabotropic glutamate receptors and diacylglycerol lipase-α initiates 2-arachidonoylglycerol formation and endocannabinoid-mediated analgesia.
Acute stress reduces pain sensitivity by engaging an endocannabinoid signaling circuit in the midbrain. The neural mechanisms governing this process and molecular identity of the endocannabinoid substance(s) involved are unknown. We combined behavior, pharmacology, immunohistochemistry, RNA interference, quantitative RT-PCR, enzyme assays, and lipidomic analyses of endocannabinoid content to uncover the role of the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG) in controlling pain sensitivity in vivo. Here, we show that footshock stress produces antinociception in rats by activating type 5 metabotropic glutamate receptors (mGlu(5)) in the dorsolateral periaqueductal gray (dlPAG) and mobilizing 2-AG. Stimulation of mGlu(5) in the dlPAG with DHPG [(S)-3,5-dihydroxyphenylglycine] triggered 2-AG formation and enhanced stress-dependent antinociception through a mechanism dependent upon both postsynaptic diacylglycerol lipase (DGL) activity, which releases 2-AG, and presynaptic CB(1) cannabinoid receptors. Pharmacological blockade of DGL activity in the dlPAG with RHC80267 [1,6-bis(cyclohexyloximinocarbonylamino)hexane] and (-)-tetrahydrolipstatin (THL), which inhibit activity of DGL-α and DGL-β isoforms, suppressed stress-induced antinociception. Inhibition of DGL activity in the dlPAG with THL selectively decreased accumulation of 2-AG without altering levels of anandamide. The putative 2-AG-synthesizing enzyme DGL-α colocalized with mGlu(5) at postsynaptic sites of the dlPAG, whereas CB(1) was confined to presynaptic terminals, consistent with a role for 2-AG as a retrograde signaling messenger. Finally, virally mediated silencing of DGL-α, but not DGL-β, transcription in the dlPAG mimicked effects of DGL inhibition in suppressing both endocannabinoid-mediated stress antinociception and 2-AG formation. The results indicate that activation of the postsynaptic mGlu(5)-DGL-α cascade triggers retrograde 2-AG signaling in vivo. This pathway is required for endocannabinoid-mediated stress-induced analgesia.
Neuroscience Program, Biomedical and Health Sciences Institute, and Psychology Department, University of Georgia, Athens, Georgia 30602-3013, USA.
Laura C
LC
Gregg
Kwang-Mook
KM
Jung
Jessica M
JM
Spradley
Rita
R
Nyilas
Richard L
RL
Suplita
Andreas
A
Zimmer
Masahiko
M
Watanabe
Ken
K
Mackie
István
I
Katona
Daniele
D
Piomelli
Andrea G
AG
Hohmann
D000698:Analgesia / Q000379:methods*; D000704:Analysis of Variance; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D063385:Cannabinoid Receptor Modulators / Q000819:agonists / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D003512:Cyclohexanones / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004565:Electroconvulsive Therapy / Q000379:methods; D063388:Endocannabinoids; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005989:Glycerides / Q000378:metabolism*; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008297:Male; D008734:Methoxyhydroxyphenylglycol / Q000008:administration & dosage / Q000031:analogs & derivatives; D051379:Mice; D016253:Microscopy, Immunoelectron; D010146:Pain / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D010487:Periaqueductal Gray / Q000187:drug effects / Q000378:metabolism; D010880:Piperidines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000627:therapeutic use; D051381:Rats; D017207:Rats, Sprague-Dawley; D043884:Receptor, Cannabinoid, CB1 / Q000378:metabolism; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism*; D000077285:Rimonabant; D013569:Synapses / Q000378:metabolism / Q000648:ultrastructure; D053719:Tandem Mass Spectrometry
D001095:Arachidonic Acids; D063385:Cannabinoid Receptor Modulators; D003512:Cyclohexanones; D063388:Endocannabinoids; D018691:Excitatory Amino Acid Antagonists; D005989:Glycerides; D010880:Piperidines; D011480:Protease Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D043884:Receptor, Cannabinoid, CB1; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate; D008734:Methoxyhydroxyphenylglycol; C121465:6-methyl-2-(phenylethynyl)pyridine; C036535:1,6-bis(cyclohexyloximinocarbonyl)hexane; C094503:glyceryl 2-arachidonate; D008071:Lipoprotein Lipase; D000077285:Rimonabant; C010117:3,4-dihydroxyphenylglycol