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Effects of synthetic cannabinoids, identified in smoked herbal products, on synaptic transmission in the brain Bela Szabo, Mario Lederer, Eszter Boros Inst. f. Pharmakologie Albert-Ludwigs-Universität Freiburg 1) Cannabinoid receptors and their typical effects in the brain Cannabinoid receptors NATURAL NH2 NH CB1 2 CB2 COOH COOH OH 9-tetrahydrocannabinol O Gi/o proteins ENDOGENOUS O OH N H neurons immune cells anandamide O OH O OH 2-arachidonoylglycerol CB1-Rezeptoren im menschlichen Gehirn: PET-Untersuchung mit [18F]MK-9470 Negative correlation: extravagance – CB1 receptor Van Laere et al., Arch Gen disorderlinessVan Laere – CB et1 al.,receptor NeuroimagePsychiatry 39: 1533-1541, 66:196-204, 20082009 2) Synthetic cannabinoids Synthetic cannabinoids from Sterling Winthrop O indomethacin pravadoline O O O N O N Cl O N O WIN55212 N O O N Ward et al., J Pharmacol Exp Ther 255: 1230-1239, 1990 O Haubrich et al., J Pharmacol Exp Ther 255: 511-522, 1990 Kuster et al., J Pharmacol Exp Ther 264: 1352-1363, 1993 Synthetic cannabinoids from John William Huffman: JWH-compounds JWH-018 JWH-210 O N O N O JWH-122 JWH-081 Aung et al., Drug Alcohol Depend 60: 133-140, 2000 Huffman JW, Cannabimimetic indoles, pyrroles, and O N O N indenes: structure–activity relationships and receptor Interactions. In: Reggio PH (ed), The Cannabinoid Receptors, The Receptors, Humana Press, 2009, pp 49- 94 3) Methods Patch-Clamp-Setup Verstärker Rollquetschpumpe Objektiv Patch-Pipette Teflonschlauch Badkammer Absaugung Hirnschnitt Begasung mit Carbogen (95% O2; 5% CO2) Becherglas mit ACSF Cerebellum: microscopic views white matter granule cell layer Purkinje cell layer molecular layer basket cell molecular layer Purkinje cell CB -R Purkinje cell 1 layer patch pipette sIPSC recording granule cell depolarisation layer 4) Synthetic cannabinoids inhibit synaptic transmission WIN55212-2 inhibits GABAergic synaptic transmission JWH-018 inhibits GABAergic synaptic transmission JWH-210 inhibits GABAergic synaptic transmission 5) Synthetic cannabinoids interfere with endocannabinoid-mediated retrograde signal transmission Retrograde signaling by cannabinoids Purkinje cell soma / dendrite mG lu 1 R 1 basket cell / q axon terminal G Ca2+ GABA-R GABA Vm endocannabinoid (2-AG) 1-R CB THC, WIN, JWH-018, JWH-210 depolarisation Depolarisation-induced suppression of inhibition (DSI) 120 depol (9P) 120 1 100 100 80 80 60 60 PRE 2 PRE sIPSC amplitude [% of PRE] sIPSC [% frequency of PRE] 050100150s 0 50 100 150 s PRE (1) 120 100 80 0.5 nA [% of PRE] DSI (2) 2 s 60 PRE sIPSC cumulative amplitude sIPSC cumulative 0 50 100 150 s JWH-210 occludes DSI depol 120 depol depol 100 SOL superfusion 80 * 60 PRE sIPSC frequency [% of PRE] 0 50 100 150 s depol 120 * depol depol 100 * JWH-210 5 x 10-6 M superfusion 80 60 PRE sIPSC [% frequency of PRE] 0 50 100 150 s Effects of synthetic cannabinoids, identified in smoked herbal products, on synaptic transmission in the brain ) The synthetic aminoalkylindole cannabinoids WIN5512-2, JHW-018 and JWH-210 inhibit GABAergic synaptic transmission in the cerebellar cortex 2) The basis of the inhibition is very likely inhibition of GABA release from the presynaptic axon terminals 3) The synthetic cannabinoids occlude endocannabinoid-mediated retrograde signal transmission between neurons, indicating that they can interfere with elementary neuronal learning basket processes cell Purkinje CB -R cell 1 WIN, JWH endocannabinoid depolarisation End.