Skip to main content
SpringerLink
Log in

Further Evidence for the Neuroplastic Role of Cannabinoids: A Study in Organotypic Hippocampal Slice Cultures

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Endocannabinoid receptors CB1R and CB2R are present in the CNS and modulate synaptic activity. By using an in vitro model, two concentrations of CB1R agonist ACEA at 0.5 and 5 μM doses and CB1R antagonist AM251 at 1 and 10 μM doses were administered in organotypic slice cultures of mouse hippocampus, and their effects on neurons and glial cells were analyzed at different time points. Exposure to low concentrations of ACEA (0.5 μM) did not seem to affect tissue organization, neuronal morphology, or glial response. In contrast, at a higher concentration of ACEA, many neurons in the dentate gyrus exhibited strong caspase-3 immunoreactivity. After treatment with AM251, we observed an increase in caspase-3 immunoreactivity and a downregulation of CB1R expression. Results show that long-term hippocampal slice cultures respond to both CB1R activation and inactivation by changing neuronal protein expression patterns. In the present study, we demonstrate that CB1R agonist ACEA promotes alterations in the neuronal cytoskeleton as well as changes in CB1R expression in organotypic hippocampal slice cultures, and that CB1R antagonist AM251 promotes neuronal death and astroglial reaction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Ameri A (1999) The effects of cannabinoids on the brain. Prog Neurobiol 58(4):315–348

  • Ashton J, Wright J, McPartland J, Tyndall J (2008) Cannabinoid CB1 and CB2 receptor ligand specificity and the development of CB2-selective agonists. Curr Med Chem 15(14):1428–1443

    Article  CAS  PubMed  Google Scholar 

  • Berghuis P, Rajnicek AM, Morozov YM, Ross RA, Mulder J, Urbán GM, Monory K, Marsicano G, Matteoli M, Canty A, Irving AJ, Katona I, Yanagawa Y, Rakic P, Lutz B, Mackie K, Harkany T (2007) Hardwiring the brain: Endocannabinoids shape neuronal connectivity. Science 316:1212–16

    Article  CAS  PubMed  Google Scholar 

  • Borner C, Holt V, Kraus J (2007) Activation of human T cells induces upregulation of cannabinoid type 1 transcription. Neuroimmunomodulation 14:281–286

    Article  PubMed  Google Scholar 

  • Brinks H, Conrad S, Vogt J, Oldekamp J, Sierra A, Deitinghoff L, Bechmann I, Alvarez-Bolado G, Heimrich B, Monnier PP, Mueller BK, Skutella T (2004) The repulsive guidance molecule RGMa is involved in the formation of afferent connections in the dentate gyrus. J Neurosci 24(15):3862–3869

    Article  CAS  PubMed  Google Scholar 

  • Brusco A, Tagliaferro PA, Saez T, Onaivi ES (2008) Ultrastructural localization of neuronal brain CB2 cannabinoid receptors. Ann N Y Acad Sci 1139:450–7

    Article  CAS  PubMed  Google Scholar 

  • Evrard S, Duhalde-Vega M, Tagliaferro P, Mirochnic S, Caltana L, Brusco A (2006) A low chronic ethanol exposure induces morphological changes in the adolescent rat brain that are not fully recovered even after a long abstinence: an immunohistochemical study. Exp Neurol 200(2):438–459

    Article  CAS  PubMed  Google Scholar 

  • Galve-Roperh I, Aguado T, Palazuelos J, Guzman M (2008) Mechanisms of control of neuron survival by the endocannabinoid system. Curr Pharm Des 14:2279–2288

    Article  CAS  PubMed  Google Scholar 

  • Gilbert GL, Kim HJ, Waataja JJ, Thayer SA (2007) Delta9-tetrahydrocannabinol protects hippocampal neurons from excitotoxicity. Brain Res 1128(1):61–9

    Article  CAS  PubMed  Google Scholar 

  • Goncalves M, Suetterlin P, Yip P, Molina Holgado F, Walker D, Oudin M, Zentar M, Pollard S, Yanez-Muñoz R, Williams G, Walsh F, Pangalos M, Doherty P (2008) A diacylglycerol lipase-CB2 cannabinoid pathway regulates adult subventricular zone neurogenesis in an age-dependent manner. Mol Cell Neurosci 38(4):526–536

    Article  CAS  PubMed  Google Scholar 

  • Grant I, Rael Cahn B (2005) Cannabis and endocannabinoid modulators: Therapeutic promises and challenges. Clin Neurosci Res 5(2-4):185–199

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Heifets B, Castillo P (2009) Endocannabinoid signaling and long-term synaptic plasticity. Annu Rev Physiol 71:283–306

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Herkenham M, Lynn AB, Little MD, Johnson MR, Melvin LS, de Costa BR, Rice KC (1990) Cannabinoid receptor localization in brain. Proc Natl Acad Sci U S A 87(5):1932–6

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hillard CJ, Manna S, Greenberg MJ, DiCamelli R, Ross RA, Stevenson LA, Murphy V, Pertwee RG, Campbell WB (1999) Synthesis and characterization of potent and selective agonists of the neuronal cannabinoid receptor (CB1). J Pharmacol Exp Ther 289(3):1427–33

    CAS  PubMed  Google Scholar 

  • Howlett AC, Barth F, Bonner TI, Cabral G, Casellas P, Devane WA, Felder CC, Herkenham M, Mackie K, Martin BR, Mechoulam R, Pertwee RG (2002) International Union of Pharmacology. XXVII. Classification of cannabinoid receptors. Pharmacol Rev 54:161–202

    Article  CAS  PubMed  Google Scholar 

  • Humar M, Graetz C, Roesslein M, Goebel U, Geiger K, Heimrich B, Pannen B (2009) Heterocyclic thioureylenes protect from calcium-dependent neuronal cell death. Mol Pharmacol 75:667–676

    Article  CAS  PubMed  Google Scholar 

  • Iversen L (2003) Cannabis and the brain. Brain 126(Pt 6):1252–1270

  • Jordan JD, He JC, Eungdamrong NJ, Gomes I, Ali W, Nguyen T, Bivona TG, Philips MR, Devi LA, Iyengar R (2005) Cannabinoid receptor induced neurite outgrowth is mediated by Rap1 activation through Galphao/I-triggered proteasomal degradation of Rap1GAPII. J Biol Chem 280(12):11413–21

    Article  CAS  PubMed  Google Scholar 

  • Katona I, Sperlágh B, Sík A, Käfalvi A, Vizi ES, Mackie K, Freund TF (1999) Presynaptically located CB1 cannabinoid receptors regulate GABA release from axon terminals of specific hippocampal interneurons. J Neurosci 19(11):4544–58

    CAS  PubMed  Google Scholar 

  • Kim D, Thayer S (2000) Activation of CB1 cannabinoid receptors inhibits neurotransmitter release from identified synaptic sites in rat hippocampal cultures. Brain Res 852:398–405

    Article  CAS  PubMed  Google Scholar 

  • Kim D, Thayer SA (2001) Cannabinoids inhibit the formation of new synapses between hippocampal neurons in culture. J Neurosci 21(10):RC146

  • Klein TW (2005) Cannabinoid-based drugs as anti-inflammatory therapeutics. Nat Rev Immunol 5:400–411

    Article  CAS  PubMed  Google Scholar 

  • Lacour P, Heimrich B, Pröls F (2007) Induction of cellular stress and chaperone activation in organotypic slice cultures of hippocampus. J Neurosci Methods 166:24–31

    Article  CAS  PubMed  Google Scholar 

  • Landucci E, Scartabelli T, Gerace E, Moroni F, Pellegrini-Giampietro D (2011) CB1 receptors and post-ischemic brain damage: Studies on the toxic and neuroprotective effects of cannabinoids in rat organotypic hippocampal slices. Neuropharmacology 60:674–682

    Article  CAS  PubMed  Google Scholar 

  • Mackie K (2005) Distribution of cannabinoid receptors in the central and peripheral nervous system. Handb Exp Pharmacol 168:299–325

    Article  CAS  PubMed  Google Scholar 

  • Mackie K (2006) Mechanisms of CB1 receptor signaling: Endocannabinoid modulation of synaptic strength. Int J Obes (Lond) 30(Suppl 1):S19–23

    Article  CAS  Google Scholar 

  • Marchalant Y, Rosi S, Wenk GL (2007) Anti-inflammatory property of the cannabinoid agonist WIN-55212-2 in a rodent model of chronic brain inflammation. Neuroscience 144:1516–1522

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mukhopadhyay B, Liu J, Osei-Hyiaman D, Godlewski G, Mukhopadhyay P, Wang L, Jeong WI, Gao B, Duester G, Mackie K, Kojima S, Kunos G (2010) Transcriptional regulation of cannabinoid receptor-1 expression in the liver by retinoic acid acting via retinoic acid receptor-gamma. J Biol Chem 285:19002–19011

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Navarrete M, Araque A (2010) Endocannabinoids potentiate synaptic transmission through stimulation of astrocytes. Neuron 68:113–126

    Article  CAS  PubMed  Google Scholar 

  • Onaivi ES, Ishiguro H, Gong JP, Patel S, Perchuk A, Meozzi PA, Myers L, Mora Z, Tagliaferro P, Gardner E, Brusco A, Akinshola BE, Liu QR, Hope B, Iwasaki S, Arinami T, Teasenfitz L, Uhl GR (2006) Discovery of the presence and functional expression of cannabinoid CB2 receptors in brain. Ann N Y Acad Sci 1074:514–536

    Article  CAS  PubMed  Google Scholar 

  • Pacher P, Bátkai S, Kunos G (2006) The endocannabinoid system as an emerging target of pharmacotherapy. Pharmacol Rev 58:389–462

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Parmentier-Batteur S, Jin K, Mao XO, Xie L, Greenberg DA (2002) Increased severity of stroke in CB1 cannabinoid receptor knock-out mice. J Neurosci 22:9771–9775

    CAS  PubMed  Google Scholar 

  • Pertwee RG (2005) Pharmacological actions of cannabinoids. Handb Exp Pharmacol (168):1-51

  • Proto MC, Gazzerro P, Di Croce L, Santoro A, Malfitano AM, Pisanti S, Laezza C, Bifulco M (2011) Interaction of endocannabinoid system and steroid hormones in the control of colon cancer cell growth. J Cell Physiol 227:250–258

    Article  Google Scholar 

  • Rubio-Araiz A, Arevalo-Martín A, Gomez-Torres O, Navarro-Galve B, Garcia-Ovejero D, Suetterlin P, Sanchez-Heras E, Molina-Holgado E, Molina-Holgado F (2008) The endocannabinoid system modulates a transient TNF pathway that induces neural stem cell proliferation. Mol Cell Neurosci 38:374–380

    Article  CAS  PubMed  Google Scholar 

  • Scallet AC (1991) Neurotoxicology of cannabis and THC: a review of chronic exposure studies in animals. Pharmacol Biochem Behav 40(3):671–6

    Article  CAS  PubMed  Google Scholar 

  • Stoppini L, Buchs P, Muller D (1991) A simple method for organotypic cultures of nervous tissue. J Neurosci Methods 37:173–182

    Article  CAS  PubMed  Google Scholar 

  • Tagliaferro P, Ramos AJ, Onaivi ES, Evrard SG, Lujilde J, Brusco A (2006) Neuronal cytoskeleton and synaptic densities are altered after a chronic treatment with the cannabinoid receptor agonist WIN 55,212-2. Brain Res 1085(1):163–76

    Article  CAS  PubMed  Google Scholar 

  • Uchigashima M, Yamazaki M, Yamasaki M, Tanimura A, Sakimura K, Kano M, Watanabe M (2011) Molecular and morphological configuration for 2-arachidonoylglycerol-mediated retrograde signaling at mossy cell-granule cell synapses in the dentate gyrus. J Neurosci 31(21):7700–14

    Article  CAS  PubMed  Google Scholar 

  • Van der Stelt M, Di Marzo V (2005) Cannabinoid receptors and their role in neuroprotection. Neuromol Med 7(1–2):37–50

    Article  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Simone Zenker for her kind technical support.

Financial Support

UBACYT 20020130300033BA (L.C.) and UBACYT 20020100100093BA (A.B...), UBACYT 20020130100258BA (A.B...), PIP CONICET 00269 (A.B...)

Author information

Authors and Affiliations

  1. Institute of Anatomy and Cell Biology, Center for Neuroscience, University of Freiburg, Freiburg, Germany

    Laura Romina Caltana & Bernd Heimrich

  2. Cell Biology and Neuroscience Institute, School of Medicine, University of Buenos Aires, Paraguay 2155, 3rd floor, Buenos Aires, 1114, Argentina

    Laura Romina Caltana & Alicia Brusco

Authors
  1. Laura Romina Caltana
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bernd Heimrich
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alicia Brusco
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Laura Romina Caltana.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Caltana, L.R., Heimrich, B. & Brusco, A. Further Evidence for the Neuroplastic Role of Cannabinoids: A Study in Organotypic Hippocampal Slice Cultures. J Mol Neurosci 56, 773–781 (2015). https://doi.org/10.1007/s12031-015-0499-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-015-0499-4

Keywords

Search

Navigation