Skip to main content
    • Aa
    • Aa
  • The International Journal of Neuropsychopharmacology, Volume 16, Issue 6
  • July 2013, pp. 1407-1419

The anxiolytic effect of cannabidiol on chronically stressed mice depends on hippocampal neurogenesis: involvement of the endocannabinoid system

  • Alline C. Campos (a1), Zaira Ortega (a2) (a3), Javier Palazuelos (a2) (a3), Manoela V. Fogaça (a1), Daniele C. Aguiar (a4), Javier Díaz-Alonso (a2) (a3), Silvia Ortega-Gutiérrez (a5), Henar Vázquez-Villa (a5), Fabricio A. Moreira (a4), Manuel Guzmán (a2) (a3), Ismael Galve-Roperh (a2) (a3) and Francisco S. Guimarães (a1)
  • DOI:
  • Published online: 09 January 2013

Cannabidiol (CBD), the main non-psychotomimetic component of the plant Cannabis sativa, exerts therapeutically promising effects on human mental health such as inhibition of psychosis, anxiety and depression. However, the mechanistic bases of CBD action are unclear. Here we investigate the potential involvement of hippocampal neurogenesis in the anxiolytic effect of CBD in mice subjected to 14 d chronic unpredictable stress (CUS). Repeated administration of CBD (30 mg/kg i.p., 2 h after each daily stressor) increased hippocampal progenitor proliferation and neurogenesis in wild-type mice. Ganciclovir administration to GFAP-thymidine kinase (GFAP-TK) transgenic mice, which express thymidine kinase in adult neural progenitor cells, abrogated CBD-induced hippocampal neurogenesis. CBD administration prevented the anxiogenic effect of CUS in wild type but not in GFAP-TK mice as evidenced in the novelty suppressed feeding test and the elevated plus maze. This anxiolytic effect of CBD involved the participation of the CB1 cannabinoid receptor, as CBD administration increased hippocampal anandamide levels and administration of the CB1–selective antagonist AM251 prevented CBD actions. Studies conducted with hippocampal progenitor cells in culture showed that CBD promotes progenitor proliferation and cell cycle progression and mimics the proliferative effect of CB1 and CB2 cannabinoid receptor activation. Moreover, antagonists of these two receptors or endocannabinoid depletion by fatty acid amide hydrolase overexpression prevented CBD-induced cell proliferation. These findings support that the anxiolytic effect of chronic CBD administration in stressed mice depends on its proneurogenic action in the adult hippocampus by facilitating endocannabinoid-mediated signalling.

Corresponding author
Address for correspondence: Dr I. Galve-Roperh, Department of Biochemistry and Molecular Biology, School of Biology, Universidad Complutense, c/Jose Antonio Novais 2, 28040 Madrid, Spain. Tel.: +34 913 944668Fax: +34 913 944672Email:
Linked references
Hide All

This list contains references from the content that can be linked to their source. For a full set of references and notes please see the PDF or HTML where available.

T Aguado , E Romero , K Monory , J Palazuelos , M Sendtner , G Marsicano , B Lutz , M Guzmán , I Galve-Roperh (2007) The CB1 cannabinoid receptor mediates excitotoxicity-induced neural progenitor proliferation and neurogenesis. J Biol Chem 282:2389223898.

MM Bergamaschi , RH Queiroz , MH Chagas , DC de Oliveira , BS De Martinis , F Kapczinski , J Quevedo , R Roesler , N Schröder , AE Nardi , R Martín-Santos , JE Hallak , AW Zuardi , JA Crippa (2011) Cannabidiol reduces the anxiety induced by simulated public speaking in treatment-naive social phobia patients. Neuropsychopharmacology 36:12191226.

A Bilkei-Gorzo , I Racz , O Valverde , M Otto , K Michel , M Sastre , A Zimmer (2005) Early age-related cognitive impairment in mice lacking cannabinoid CB1 receptors. Proc Natl Acad Sci USA 102:1567015675.

T Bisogno , L Hanus , L De Petrocellis , S Tchilibon , DE Ponde , I Brandi , AS Moriello , JB Davis , R Mechoulam , V Di Marzo (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 134:845852.

RM Bitencourt , FA Pamplona , RN Takahashi (2008) Facilitation of contextual fear memory extinction and anti-anxiogenic effects of AM404 and cannabidiol in conditioned rats. Eur Neuropsychopharmacol 18:849859.

A Busquets-Garcia , E Puighermanal , A Pastor , R de la Torre , R Maldonado , A Ozaita (2011) Differential role of anandamide and 2-arachidonoylglycerol in memory and anxiety-like responses. Biol Psychiatry 70:479486.

AC Campos , FS Guimaraes (2008) Involvement of 5HT1A receptors in the anxiolytic-like effects of cannabidiol injected into the dorsolateral periaqueductal gray of rats. Psychopharmacology (Berl) 199:223230.

AC Campos , FS Guimaraes (2009) Evidence for a potential role for TRPV1 receptors in the dorsolateral periaqueductal gray in the attenuation of the anxiolytic effects of cannabinoids. Prog Neuropsychopharmacol Biol Psychiatry 33:15171521.

PC Casarotto , FV Gomes , LB Resstel , FS Guimaraes (2010) Cannabidiol inhibitory effect on marble-burying behaviour: involvement of CB1 receptors. Behav Pharmacol 21:353358.

A Castillo , MR Tolón , J Fernández-Ruiz , J Romero , J Martinez-Orgado (2010) The neuroprotective effect of cannabidiol in an in vitro model of newborn hypoxic-ischemic brain damage in mice is mediated by CB(2) and adenosine receptors. Neurobiol Dis 37:434440.

DJ David , BA Samuels , Q Rainer , JW Wang , D Marsteller , I Mendez , M Drew , DA Craig , BP Guiard , JP Guilloux , RP Artymyshyn , AM Gardier , C Gerald , IA Antonijevic , ED Leonardo , R Hen (2009) Neurogenesis-dependent and -independent effects of fluoxetine in an animal model of anxiety/depression. Neuron 62:479493.

DJ David , J Wang , BA Samuels , Q Rainer , I David , AM Gardier , R Hen (2010) Implications of the functional integration of adult-born hippocampal neurons in anxiety-depression disorders. Neuroscientist 16:578591.

S Deiana , A Watanabe , Y Yamasaki , N Amada , M Arthur , S Fleming , H Woodcock , P Dorward , B Pigliacampo , S Close , B Platt , G Riedel (2012) Plasma and brain pharmacokinetic profile of cannabidiol (CBD), cannabidivarine (CBDV), Delta(9)-tetrahydrocannabivarin (THCV) and cannabigerol (CBG) in rats and mice following oral and intraperitoneal administration and CBD action on obsessive-compulsive behaviour. Psychopharmacology (Berl) 219:859873.

W Deng , JB Aimone , FH Gage (2010) New neurons and new memories: how does adult hippocampal neurogenesis affect learning and memory? Nat Rev Neurosci 11:339350.

A Dranovsky , AM Picchini , T Moadel , AC Sisti , A Yamada , S Kimura , ED Leonardo , R Hen (2011) Experience dictates stem cell fate in the adult hippocampus. Neuron 70:908923.

G Esposito , C Scuderi , M Valenza , GI Togna , V Latina , D De Filippis , M Cipriano , MR Carratù , T Iuvone , L Steardo (2011) Cannabidiol reduces Abeta-induced neuroinflammation and promotes hippocampal neurogenesis through PPARgamma involvement. PLoS ONE 6:e28668.

P Fadda , L Robinson , W Fratta , RG Pertwee , G Riedel (2004) Differential effects of THC- or CBD-rich cannabis extracts on working memory in rats. Neuropharmacology 47:11701179.

I Galve-Roperh , J Palazuelos , T Aguado , M Guzman (2009) The endocannabinoid system and the regulation of neural development: potential implications in psychiatric disorders. Eur Arch Psychiatry Clin Neurosci 259:371382.

AD Garcia , NB Doan , T Imura , TG Bush , MV Sofroniew (2004) GFAP-expressing progenitors are the principal source of constitutive neurogenesis in adult mouse forebrain. Nat Neurosci 7:12331241.

FV Gomes , LB Resstel , FS Guimaraes (2011) The anxiolytic-like effects of cannabidiol injected into the bed nucleus of the stria terminalis are mediated by 5-HT1A receptors. Psychopharmacology (Berl) 213:465473.

MB Goncalves , P Suetterlin , P Yip , F Molina-Holgado , DJ Walker , MJ Oudin , MP Zentar , S Pollard , RJ Yáñez-Muñoz , G Williams , FS Walsh , MN Pangalos , P Doherty (2008) A diacylglycerol lipase-CB2 cannabinoid pathway regulates adult subventricular zone neurogenesis in an age-dependent manner. Mol Cell Neurosci 38:526536.

FS Guimaraes , TM Chiaretti , FG Graeff , AW Zuardi (1990) Anti-anxiety effect of cannabidiol in the elevated plus-maze. Psychopharmacology (Berl) 100:558559.

BD Heifets , PE Castillo (2009) Endocannabinoid signaling and long-term synaptic plasticity. Annu Rev Physiol 71:283306.

MN Hill , CJ Hillard , FR Bambico , S Patel , BB Gorzalka , G Gobbi (2009) The therapeutic potential of the endocannabinoid system for the development of a novel class of antidepressants. Trends Pharmacol Sci 30:484493.

MN Hill , JS Kambo , JC Sun , BB Gorzalka , LA Galea (2006) Endocannabinoids modulate stress-induced suppression of hippocampal cell proliferation and activation of defensive behaviours. Eur J Neurosci 24:18451849.

BL Hungund , KY Vinod , SA Kassir , BS Basavarajappa , R Yalamanchili , TB Cooper , JJ Mann , V Arango (2004) Upregulation of CB1 receptors and agonist-stimulated [35S]GTPgammaS binding in the prefrontal cortex of depressed suicide victims. Mol Psychiatry 9:184190.

AA Izzo , F Borrelli , R Capasso , V Di Marzo , R Mechoulam (2009) Non-psychotropic plant cannabinoids: new therapeutic opportunities from an ancient herb. Trends Pharmacol Sci 30:515527.

W Jiang , Y Zhang , L Xiao , J Van Cleemput , SP Ji , G Bai , X Zhang (2005) Cannabinoids promote embryonic and adult hippocampus neurogenesis and produce anxiolytic- and antidepressant-like effects. J Clin Invest 115:31043116.

S Kathuria , S Gaetani , D Fegley , F Valiño , A Duranti , A Tontini , M Mor , G Tarzia , G La Rana , A Calignano , A Giustino , M Tattoli , M Palmery , V Cuomo , D Piomelli (2003) Modulation of anxiety through blockade of anandamide hydrolysis. Nat Med 9:7681.

SG Kinsey , ST O'Neal , JZ Long , BF Cravatt , AH Lichtman (2011) Inhibition of endocannabinoid catabolic enzymes elicits anxiolytic-like effects in the marble burying assay. Pharmacol Biochem Behav 98:2127.

DL Krebs-Kraft , MN Hill , CJ Hillard , MM McCarthy (2010) Sex difference in cell proliferation in developing rat amygdala mediated by endocannabinoids has implications for social behavior. Proc Natl Acad Sci USA 107:2053520540.

FM Leweke , D Piomelli , F Pahlisch , D Muhl , CW Gerth , C Hoyer , J Klosterkötter , M Hellmich , D Koethe (2012) Cannabidiol enhances anandamide signaling and alleviates psychotic symptoms of schizophrenia. Transl Psychiatry 2:e94.

Y Marchalant , HM Brothers , GJ Norman , K Karelina , AC DeVries , GL Wenk (2009a) Cannabinoids attenuate the effects of aging upon neuroinflammation and neurogenesis. Neurobiol Dis 34:300307.

Y Marchalant , HM Brothers , GL Wenk (2009b) Cannabinoid agonist WIN-55,212-2 partially restores neurogenesis in the aged rat brain. Mol Psychiatry 14:10681069.

J Mulder , T Aguado , E Keimpema , K Barabás , CJ Ballester Rosado , L Nguyen , K Monory , G Marsicano , V Di Marzo , YL Hurd , F Guillemot , K Mackie , B Lutz , M Guzmán , HC Lu , I Galve-Roperh , T Harkany (2008) Endocannabinoid signaling controls pyramidal cell specification and long-range axon patterning. Proc Natl Acad Sci USA 105:87608765.

J Palazuelos , T Aguado , A Egia , R Mechoulam , M Guzmán , I Galve-Roperh (2006) Non-psychoactive CB2 cannabinoid agonists stimulate neural progenitor proliferation. FASEB J 20:24052407.

J Palazuelos , T Aguado , MR Pazos , B Julien , C Carrasco , E Resel , O Sagredo , C Benito , J Romero , I Azcoitia , J Fernández-Ruiz , M Guzmán , I Galve-Roperh (2009) Microglial CB2 cannabinoid receptors are neuroprotective in Huntington's disease excitotoxicity. Brain 132:31523164.

J Palazuelos , Z Ortega , J Díaz-Alonso , M Guzmán , I Galve-Roperh (2011) CB2 cannabinoid receptors promote neural progenitor cell proliferation via mTORC1 signaling. J Biol Chem 287:11981209.

VK Parihar , B Hattiangady , R Kuruba , B Shuai , AK Shetty (2011) Predictable chronic mild stress improves mood, hippocampal neurogenesis and memory. Mol Psychiatry 16:171183.

S Patel , CJ Hillard (2008) Adaptations in endocannabinoid signaling in response to repeated homotypic stress: a novel mechanism for stress habituation. Eur J Neurosci 27:28212829.

RG Pertwee , AC Howlett , ME Abood , SP Alexander , V Di Marzo , MR Elphick , PJ Greasley , HS Hansen , G Kunos , K Mackie , R Mechoulam , RA Ross (2010) International union of basic and clinical pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond CB and CB. Pharmacol Rev 62:588631.

JM Revest , D Dupret , M Koehl , C Funk-Reiter , N Grosjean , PV Piazza , DN Abrous (2009) Adult hippocampal neurogenesis is involved in anxiety-related behaviors. Mol Psychiatry 14:959967.

A Sahay , KN Scobie , AS Hill , CM O'Carroll , MA Kheirbek , NS Burghardt , AA Fenton , A Dranovsky , R Hen (2011) Increasing adult hippocampal neurogenesis is sufficient to improve pattern separation. Nature 472:466470.

L Santarelli , M Saxe , C Gross , A Surget , F Battaglia , S Dulawa , N Weisstaub , J Lee , R Duman , O Arancio , C Belzung , R Hen (2003) Requirement of hippocampal neurogenesis for the behavioral effects of antidepressants. Science 301:805809.

RJ Schloesser , M Lehmann , K Martinowich , HK Manji , M Herkenham (2010) Environmental enrichment requires adult neurogenesis to facilitate the recovery from psychosocial stress. Mol Psychiatry 15:11521163.

JS Snyder , A Soumier , M Brewer , J Pickel , HA Cameron (2011) Adult hippocampal neurogenesis buffers stress responses and depressive behaviour. Nature 476:458461.

A Surget , A Tanti , ED Leonardo , A Laugeray , Q Rainer , C Touma , R Palme , G Griebel , Y Ibarguen-Vargas , R Hen , C Belzung (2011) Antidepressants recruit new neurons to improve stress response regulation. Mol Psychiatry 16:11771188.

A Thomas , GL Baillie , AM Phillips , RK Razdan , RA Ross , RG Pertwee (2007) Cannabidiol displays unexpectedly high potency as an antagonist of CB1 and CB2 receptor agonists in vitro. Br J Pharmacol 150:613623.

SA Wolf , A Bick-Sander , K Fabel , P Leal-Galicia , S Tauber , G Ramirez-Rodriguez , A Müller , A Melnik , TP Waltinger , O Ullrich , G Kempermann (2010) Cannabinoid receptor CB1 mediates baseline and activity-induced survival of new neurons in adult hippocampal neurogenesis. Cell Commun Signal 8:12.

TV Zanelati , C Biojone , FA Moreira , FS Guimarães , SR Joca (2010) Antidepressant-like effects of cannabidiol in mice: possible involvement of 5-HT1A receptors. Br J Pharmacol 159:122128.

Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

The International Journal of Neuropsychopharmacology
  • ISSN: 1461-1457
  • EISSN: 1469-5111
  • URL: /core/journals/the-international-journal-of-neuropsychopharmacology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *