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The anxiolytic-like effects of cannabidiol injected into the bed nucleus of the stria terminalis are mediated by 5-HT1A receptors

Abstract

Rationale

Cannabidiol (CBD) is a non-psychotomimetic compound from Cannabis sativa that induces anxiolytic-like effects in rodents and humans after systemic administration. Previous results from our group showed that CBD injection into the bed nucleus of the stria terminalis (BNST) attenuates conditioned aversive responses. The aim of this study was to further investigate the role of this region on the anxiolytic effects of the CBD. Moreover, considering that CBD can activate 5-HT1A receptors, we also verified a possible involvement of these receptors in those effects.

Methods

Male Wistar rats received injections of CBD (15, 30, or 60 nmol) into the BNST and were exposed to the elevated plus-maze (EPM) or to the Vogel conflict test (VCT), two widely used animal models of anxiety.

Results

CBD increased open arms exploration in the EPM as well as the number of punished licks in the VCT, suggesting an anxiolytic-like effect. The drug did not change the number of entries into the enclosed arms of the EPM nor interfered with water consumption or nociceptive threshold, discarding potential confounding factors in the two tests. Moreover, pretreatment with the 5-HT1A receptor antagonist WAY100635 (0.37 nmol) blocked the effects of CBD in both models.

Conclusions

These results give further support to the proposal that BNST is involved in the anxiolytic-like effects of CBD observed after systemic administration, probably by facilitating local 5-HT1A receptor-mediated neurotransmission.

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References

  1. Alheid GF (2003) Extended amygdala and basal forebrain. Ann NY Acad Sci 985:185–205

  2. Alves FHF, Crestani CC, Gomes FV, Guimaraes FS, Correa FMA, Resstel LBM (2010) Cannabidiol injected into the bed nucleus of the stria terminalis modulates the baroreflex activity through 5-HT1A receptors. Pharmacol Res 62:228–236

  3. Barnes NM, Sharp T (1999) A review of central 5-HT receptors and their function. Neuropharmacology 38:1083–1152

  4. Bisogno T, Hanus L, De Petrocellis L, Tchilibon S, Ponde DE, Brandi I, Moriello AS, Davis JB, Mechoulam R, Di Marzo V (2001) Molecular targets for cannabidiol and its synthetic analogues: effect on vanilloid VR1 receptors and on the cellular uptake and enzymatic hydrolysis of anandamide. Br J Pharmacol 134:845–852

  5. Bosker FJ, Klompmakers A, Westenberg HGM (1997) Postsynaptic 5-HT1A receptors mediate 5-hydroxytryptamine release in the amygdala through a feedback to the caudal linear raphe. Eur J Pharmacol 333:147–157

  6. Campos AC, Guimaraes FS (2008) Involvement of 5HT1A receptors in the anxiolytic-like effects of cannabidiol injected into the dorsolateral periaqueductal gray of rats. Psychopharmacol (Berl) 199:223–230

  7. Carobrez AP, Bertoglio LJ (2005) Ethological and temporal analyses of anxiety-like behavior: the elevated plus-maze model 20 years on. Neurosci Biobehav Rev 29:1193–1205

  8. Casada JH, Dafny N (1991) Restraint and stimulation of bed nucleus of the stria terminalis produce similar stress-like behaviors. Brain Res Bull 27:207–212

  9. Chalmers DT, Watson SJ (1991) Comparative anatomical distribution of 5-HT1A receptor mRNA and 5-HT1A binding in rat brain—a combined in situ hybridisation/in vitro receptor autoradiographic study. Brain Res 561:51–60

  10. Commons KG, Connolley KR, Valentino RJ (2003) A neurochemically distinct dorsal raphe-limbic circuit with a potential role in affective disorders. Neuropsychopharmacology 28:206–215

  11. Crippa JA, Zuardi AW, Garrido GE, Wichert-Ana L, Guarnieri R, Ferrari L, Azevedo-Marques PM, Hallak JE, McGuire PK, Filho Busatto G (2004) Effects of cannabidiol (CBD) on regional cerebral blood flow. Neuropsychopharmacology 29:417–426

  12. Davis M, Walker DL, Miles L, Grillon C (2010) Phasic vs sustained fear in rats and humans: role of the extended amygdala in fear vs anxiety. Neuropsychopharmacology 35:105–135

  13. Dunn JD (1987) Plasma corticosterone responses to electrical stimulation of the bed nucleus of the stria terminalis. Brain Res 407:327–331

  14. Dunn JD, Williams TJ (1995) Cardiovascular responses to electrical stimulation of the bed nucleus of the stria terminalis. J Comp Neurol 352:227–234

  15. Engel JA, Hjorth S, Svensson K, Carlsson A, Liljequist S (1984) Anticonflict effect of the putative serotonin receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). Eur J Pharmacol 105:365–368

  16. Fusar-Poli P, Crippa JA, Bhattacharyya S, Borgwardt SJ, Allen P, Martin-Santos R, Seal M, Surguladze SA, O'Carrol C, Atakan Z, Zuardi AW, McGuire PK (2009) Distinct effects of {delta}9-tetrahydrocannabinol and cannabidiol on neural activation during emotional processing. Arch Gen Psychiatry 66:95–105

  17. Griebel G, Rodgers RJ, Perrault G, Sanger DJ (1999) Behavioural profiles in the mouse defence test battery suggest anxiolytic potential of 5-HT1A receptor antagonists. Psychopharmacol (Berl) 144:121–130

  18. Griebel G, Rodgers RJ, Perrault G, Sanger DJ (2000) The effects of compounds varying in selectivity as 5-HT1A receptor antagonists in three rat models of anxiety. Neuropharmacology 39:1848–1857

  19. Grlie L (1976) A comparative study on some chemical and biological characteristics of various samples of cannabis resin. Bull Narc 14:37–46

  20. Gross C, Zhuang X, Stark K, Ramboz S, Oosting R, Kirby L, Santarelli L, Beck S, Hen R (2002) Serotonin1A receptor acts during development to establish normal anxiety-like behaviour in the adult. Nature 416:396–400

  21. Guimaraes FS, Chiaretti TM, Graeff FG, Zuardi AW (1990) Antianxiety effect of cannabidiol in the elevated plus-maze. Psychopharmacol (Berl) 100:558–559

  22. Guo JD, Hammack SE, Hazra R, Levita L, Rainnie DG (2009) Bi-directional modulation of bed nucleus of stria terminalis neurons by 5-HT: molecular expression and functional properties of excitatory 5-HT receptor subtypes. Neuroscience 164:1776–1793

  23. Heidmann DE, Szot P, Kohen R, Hamblin MW (1998) Function and distribution of three rat 5-hydroxytryptamine7 (5-HT7) receptor isoforms produced by alternative splicing. Neuropharmacology 37:1621–1632

  24. Heisler LK, Chu HM, Brennan TJ, Danao JA, Bajwa P, Parsons LH, Tecott LH (1998) Elevated anxiety and antidepressant-like responses in serotonin 5-HT1A receptor mutant mice. Proc Natl Acad Sci USA 95:15049–15054

  25. Izzo AA, Borrelli F, Capasso R, Di Marzo V, Mechoulam R (2009) Non-psychotropic plant cannabinoids: new therapeutic opportunities from an ancient herb. Trends Pharmacol Sci 30:515–527

  26. Karniol IG, Shirakawa I, Kasinski N, Pfeferman A, Carlini EA (1974) Cannabidiol interferes with the effects of delta 9—tetrahydrocannabinol in man. Eur J Pharmacol 28:172–177

  27. Lemos JI, Resstel LB, Guimaraes FS (2010) Involvement of the prelimbic prefrontal cortex on cannabidiol-induced attenuation of contextual conditioned fear in rats. Behav Brain Res 207:105–111

  28. Levita L, Hammack SE, Mania I, Li XY, Davis M, Rainnie DG (2004) 5-Hydroxytryptamine1A-like receptor activation in the bed nucleus of the stria terminalis: electrophysiological and behavioral studies. Neuroscience 128:583–596

  29. Lowry CA, Hale MW, Evans AK, Heerkens J, Staub DR, Gasser PJ, Shekhar A (2008) Serotonergic systems, anxiety, and affective disorder: focus on the dorsomedial part of the dorsal raphe nucleus. Ann NY Acad Sci 1148:86–94

  30. Mengod G, Nguyen H, Le H, Waeber C, Lubbert H, Palacios JM (1990) The distribution and cellular localization of the serotonin 1C receptor mRNA in the rodent brain examined by in situ hybridization histochemistry. Comparison with receptor binding distribution. Neuroscience 35:577–591

  31. Millan MJ (2003) The neurobiology and control of anxious states. Prog Neurobiol 70:83–244

  32. Millan MJ, Brocco M (2003) The Vogel conflict test: procedural aspects, gamma-aminobutyric acid, glutamate and monoamines. Eur J Pharmacol 463:67–96

  33. Moreira FA, Aguiar DC, Guimaraes FS (2006) Anxiolytic-like effect of cannabidiol in the rat Vogel conflict test. Prog Neuropsychopharmacol Biol Psychiatry 30:1466–1471

  34. Parks CL, Robinson PS, Sibille E, Shenk T, Toth M (1998) Increased anxiety of mice lacking the serotonin1A receptor. Proc Natl Acad Sci USA 95:10734–10739

  35. Paxinos G, Watson C (1997) The rat brain in stereotaxic coordinates, 2nd edn. Academic, Sydney

  36. Petitet F, Jeantaud B, Reibaud M, Imperato A, Dubroeucq MC (1998) Complex pharmacology of natural cannabinoids: evidence for partial agonist activity of delta9-tetrahydrocannabinol and antagonist activity of cannabidiol on rat brain cannabinoid receptors. Life Sci 63:PL1–PL6

  37. Phelix CF, Liposits Z, Paull WK (1992) Serotonin–CRF interaction in the bed nucleus of the stria terminalis: a light microscopic double-label immunocytochemical analysis. Brain Res Bull 28:943–948

  38. Pompeiano M, Palacios JM, Mengod G (1994) Distribution of the serotonin 5-HT2 receptor family mRNAs: comparison between 5-HT2A and 5-HT2C receptors. Brain Res Mol Brain Res 23:163–178

  39. Rainnie DG (1999) Neurons of the bed nucleus of the stria terminalis (BNST). Electrophysiological properties and their response to serotonin. Ann NY Acad Sci 877:695–699

  40. Ramboz S, Oosting R, Amara DA, Kung HF, Blier P, Mendelsohn M, Mann JJ, Brunner D, Hen R (1998) Serotonin receptor 1A knockout: an animal model of anxiety-related disorder. Proc Natl Acad Sci USA 95:14476–14481

  41. Resstel LB, Joca SR, Moreira FA, Correa FM, Guimaraes FS (2006) Effects of cannabidiol and diazepam on behavioral and cardiovascular responses induced by contextual conditioned fear in rats. Behav Brain Res 172:294–298

  42. Resstel LB, Alves FH, Reis DG, Crestani CC, Correa FM, Guimaraes FS (2008) Anxiolytic-like effects induced by acute reversible inactivation of the bed nucleus of stria terminalis. Neuroscience 154:869–876

  43. Resstel LB, Tavares RF, Lisboa SF, Joca SR, Correa FM, Guimaraes FS (2009) 5-HT1A receptors are involved in the cannabidiol-induced attenuation of behavioural and cardiovascular responses to acute restraint stress in rats. Br J Pharmacol 156:181–188

  44. Rickels K, Rynn M (2002) Pharmacotherapy of generalized anxiety disorder. J Clin Psychiatry 63 (Suppl 14):9–16

  45. Russo EB, Burnett A, Hall B, Parker KK (2005) Agonistic properties of cannabidiol at 5-HT1a receptors. Neurochem Res 30:1037–1043

  46. Soares VP, Campos AC, Bortoli VC, Zangrossi H Jr, Guimaraes FS, Zuardi AW (2010) Intra-dorsal periaqueductal gray administration of cannabidiol blocks panic-like response by activating 5-HT1A receptors. Behav Brain Res 213:225–229

  47. Thomas BF, Gilliam AF, Burch DF, Roche MJ, Seltzman HH (1998) Comparative receptor binding analyses of cannabinoid agonists and antagonists. J Pharmacol Exp Ther 285:285–292

  48. Treit D, Aujla H, Menard J (1998) Does the bed nucleus of the stria terminalis mediate fear behaviors? Behav Neurosci 112:379–386

  49. Verge D, Daval G, Patey A, Gozlan H, el Mestikawy S, Hamon M (1985) Presynaptic 5-HT autoreceptors on serotonergic cell bodies and/or dendrites but not terminals are of the 5-HT1A subtype. Eur J Pharmacol 113:463–464

  50. Waddell J, Morris RW, Bouton ME (2006) Effects of bed nucleus of the stria terminalis lesions on conditioned anxiety: aversive conditioning with long-duration conditional stimuli and reinstatement of extinguished fear. Behav Neurosci 120:324–336

  51. Walker DL, Toufexis DJ, Davis M (2003) Role of the bed nucleus of the stria terminalis versus the amygdala in fear, stress, and anxiety. Eur J Pharmacol 463:199–216

  52. Xu T, Pandey SC (2000) Cellular localization of serotonin(2A) (5HT(2A)) receptors in the rat brain. Brain Res Bull 51:499–505

  53. Zanelati TV, Biojone C, Moreira FA, Guimaraes FS, Joca SR (2010) Antidepressant-like effects of cannabidiol in mice: possible involvement of 5-HT1A receptors. Br J Pharmacol 159:122–128

  54. Zangrossi H Jr, Viana MB, Graeff FG (1999) Anxiolytic effect of intra-amygdala injection of midazolam and 8-hydroxy-2(di-n-propylamino)tetralin in the elevated T-maze. Eur J Pharmacol 369:267–270

  55. Zanoveli JM, Nogueira RL, Zangrossi H Jr (2003) Serotonin in the dorsal periaqueductal gray modulates inhibitory avoidance and one-way escape behaviors in the elevated T-maze. Eur J Pharmacol 473:153–161

  56. Zuardi AW (2008) Cannabidiol: from an inactive cannabinoid to a drug with wide spectrum of action. Rev Bras Psiquiatr 30:271–280

  57. Zuardi AW, Shirakawa I, Finkelfarb E, Karniol IG (1982) Action of cannabidiol on the anxiety and other effects produced by delta 9-THC in normal subjects. Psychopharmacol (Berl) 76:245–250

  58. Zuardi AW, Cosme RA, Graeff FG, Guimaraes FS (1993) Effects of ipsapirone and cannabidiol on human experimental anxiety. J Psychopharmacol 7:82–88

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Acknowledgements

The authors thank THC Pharm for having kindly donated CBD; Ivanilda A.C. Fortunato and J.C. de Aguiar for technical support. FV Gomes has a CNPq MsC fellowship (130171/2009-3). This research was also supported by grants from FAPESP (2009/03187-9 and 2007/03685-3), CNPq (480550/2007-7 and 305996/2008-8), and FAEPA.

Conflicts of interest

The authors declare no conflicts of interest.

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Correspondence to Francisco S. Guimarães.

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Gomes, F.V., Resstel, L.B.M. & Guimarães, F.S. The anxiolytic-like effects of cannabidiol injected into the bed nucleus of the stria terminalis are mediated by 5-HT1A receptors. Psychopharmacology 213, 465–473 (2011). https://doi.org/10.1007/s00213-010-2036-z

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Keywords

  • Cannabinoids
  • Anxiety
  • Serotonin receptor
  • Rat
  • Maze
  • Limbic system
  • Behavior
  • Anxiolytic
  • Animal model