Skip to main content
SpringerLink
Log in

Opposing roles of dorsomedial hypothalamic CB1 and TRPV1 receptors in anandamide signaling during the panic-like response elicited in mice by Brazilian rainbow Boidae snakes

  • Original Investigation
  • Published:
Psychopharmacology Aims and scope Submit manuscript

Abstract

Rationale

The endocannabinoid system plays an important role in the organization of panic-like defensive behavior. Threatening situations stimulate brain areas, such as the dorsomedial hypothalamus (DMH). However, there is a lack of studies addressing the role of the DMH endocannabinoid system in panic-like responses.

Objectives

We aimed to verify which mechanisms underlie anandamide-mediated responses in the DMH.

Methods

To test the hypothesis that the anandamide produces panicolytic-like effects, we treated mice with intra-DMH microinjections of vehicle or increasing doses of anandamide (0.5, 5, or 50 pmol) and then performed confrontation with the South American snake Epicrates cenchria assisi.

Results

Intra-DMH anandamide treatment yielded a U-shaped dose-response curve with no effect of the lowest (0.5 pmol) or the highest (50 pmol) dose and significant inhibition of panic-like responses at the intermediate (5 pmol) dose. In addition, this panicolytic-like effect was prevented by pretreatment of the DMH with the CB1 receptor antagonist AM251 (100 pmol). However, pretreatment of the DMH with the TRPV1 receptor antagonist 6-iodo-nordihydrocapsaicin (3 nmol) restored the panicolytic-like effect of the highest dose of anandamide. Immunohistochemistry revealed that CB1 receptors were present primarily on axonal fibers, while TRPV1 receptors were found almost exclusively surrounding the perikarya in DMH.

Conclusions

The present results suggest that anandamide exerts a panicolytic-like effect in the DMH by activation of CB1 receptors and that TRPV1 receptors are related to the lack of effect of the highest dose of anandamide.

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

  • Aguiar DC, Almeida-Santos AF, Moreira FA, Guimarães FS (2015) Involvement of TRPV1 channels in the periaqueductal grey on the modulation of innate fear responses. Acta Neuropsychiatr 27:97–105

    Article  PubMed  Google Scholar 

  • Almada RC, Coimbra NC (2015) Recruitment of striatonigral disinhibitory and nigrotectal inhibitory GABAergic pathways during the organization of defensive behavior by mice in a dangerous environment with the venomous snake Bothrops alternatus (Reptilia, Viperidae). Synapse 69:299–313

  • Almada RC, Roncon CM, Elias-Filho DH, Coimbra NC (2015) Endocannabinoid signaling mechanisms in the substantia nigra pars reticulata modulate GABAergic nigrotectal pathways in mice threatened by urutu-cruzeiro venomous pit viper. Neuroscience 303:503–514

  • Bard P (1928) A diencephalic mechanism for the expression of rage with special reference to the sympathetic nervous system. Am J Phys 84:490–515

    Article  Google Scholar 

  • Batista LA, Bastos JR, Moreira FA (2015) Role of endocannabinoid signalling in the dorsolateral periaqueductal grey in the modulation of distinct panic-like responses. J Psychopharmacol 29:335–343

    Article  CAS  PubMed  Google Scholar 

  • Biagioni AF, de Oliveira RC, de Oliveira R, da Silva JA, dos Anjos-Garcia T, Roncon CM, Corrado AP, Zangrossi HJ, Coimbra NC (2016a) 5-Hydroxytryptamine 1A receptors in the dorsomedial hypothalamus connected to dorsal raphe nucleus inputs modulate defensive behaviours and mediate innate fear-induced antinociception. Eur Neuropsychopharmacol 26:532–545

    Article  CAS  PubMed  Google Scholar 

  • Biagioni AF, dos Anjos-Garcia T, Ullah F, Fisher IR, Falconi-Sobrinho LL, de Freitas RL, Felippotti TT, Coimbra NC (2016b) Neuroethological validation of an experimental apparatus to evaluate oriented and non-oriented escape behaviours: comparison between the polygonal arena with a burrow and the circular enclosure of an open-field test. Behav Brain Res 298(Part B):65–77

    Article  PubMed  Google Scholar 

  • Blanchard DC, Griebel G, Blanchard RJ (2001) Mouse defensive behaviors: pharmacological and behavioral assays for anxiety and panic. Neurosci Biobehav Rev 25:205–218

    Article  CAS  PubMed  Google Scholar 

  • Borelli KG, Nobre MJ, Brandão ML, Coimbra NC (2004) Effects of acute and chronic fluoxetine and diazepam on freezing behavior induced by electrical stimulation of dorsolateral and lateral columns of the periaqueductal gray matter. Pharmacol Biochem Behav 77:557–566

    Article  CAS  PubMed  Google Scholar 

  • Bortolato M, Campolongo P, Mangieri RA, Scattoni ML, Frau R, Trezza V, La Rana G, Russo R, Calignano A, Gessa GL, Cuomo V, Piomelli D (2006) Anxiolytic-like properties of the anandamide transport inhibitor AM404. Neuropsychopharmacol 31:2652–2659

    Article  CAS  Google Scholar 

  • Calvo F, Lobão-Soares B, de Freitas RL, Paschoalin-Maurin T, dos Anjos-Garcia T, Medeiros P, da Silva JA, Lovick TA, Coimbra NC (2019) The endogenous opioid system modulates defensive behavior evoked by Crotalus durissus terrificus: panicolytic-like effect of intracollicular non-selective opioid receptors blockade. J Psychopharmacol 33:51–61

  • Campos AC, Guimarães FS (2009) Evidence for a potential role for TRPV1 receptors in the dorsolateral periaqueductal gray in the attenuation of the anxiolytic effects of cannabinoids. Prog Neuro-Psychopharmacol Biol Psychiatry 33:1517–1521

    Article  CAS  Google Scholar 

  • Casarotto PC, Terzian ALB, Aguiar DC, Zangrossi H, Guimarães FS, Wotjak CT, Moreira FA (2012) Opposing roles for cannabinoid receptor type-1 (CB1) and transient receptor potential vanilloid type-1 channel (TRPV1) on the modulation of panic-like responses in rats. Neuropsychopharmacology 37:478–486

    Article  CAS  PubMed  Google Scholar 

  • Coimbra NC, Leão-Borges PC, Brandao ML (1989) GABAergic fibers from substantia nigra pars reticulata modulate escape behavior induced by midbrain central gray stimulation. Braz J Med Biol Res 22:111–114

    CAS  PubMed  Google Scholar 

  • Coimbra NC, Paschoalin-Maurin T, Bassi GS, Kanashiro A, Biagioni AF, Felippotti TT, Elias-Filho DH, Mendes-Gomes J, Cysne-Coimbra JP, Almada RC, Lobão-Soares B (2017) Critical neuropsychobiological analysis of panic attack- and anticipatory anxiety-like behaviors in rodents confronted with snakes in polygonal arenas and complex labyrinths: a comparison to the elevated plus- and T-maze behavioral tests. Braz J Psychiatr 39:72–83

    Article  Google Scholar 

  • Cristino L, de Petrocellis L, Pryce G, Baker D, Guglielmotti V, Di Marzo V (2006) Immunohistochemical localization of cannabinoid type 1 and vanilloid transient receptor potential vanilloid type 1 receptors in the mouse brain. Neuroscience 139:1405–1415

    Article  CAS  PubMed  Google Scholar 

  • de Bortoli VC, Yamashita PSM, Zangrossi H Jr (2013) 5-HT1A and 5-HT2A receptor control of a panic-like defensive response in the rat dorsomedial hypothalamic nucleus. J Psychopharmacol 27:1116–1123

    Article  CAS  PubMed  Google Scholar 

  • dos Anjos-Garcia T, Ullah F, Falconi-Sobrinho LL, Coimbra NC (2017) CB1 cannabinoid receptor-mediated anandamide signalling reduces the defensive behaviour evoked through GABAA receptor blockade in the dorsomedial division of the ventromedial hypothalamus. Neuropharmacology 113(Part A):156–166

    Article  CAS  PubMed  Google Scholar 

  • Egertová M, Giang DK, Cravatt BF, Elphick MR (1998) A new perspective on cannabinoid signalling: complementary localization of fatty acid amide hydrolase and the CB1 receptor in rat brain. Proc Biol Sci 265:2081–2085

    Article  PubMed  PubMed Central  Google Scholar 

  • Elokely K, Velisetty P, Delemotte L, Palovcak E, Klein ML, Rohacs T, Carnevale V (2016) Understanding TRPV1 activation by ligands: insights from the binding modes of capsaicin and resiniferatoxin. Proc Natl Acad Sci U S A 113:E137–E145

    Article  CAS  PubMed  Google Scholar 

  • Finn DP, Jhaveri MD, Beckett SR, Roe CH, Kendall DA, Marsden CA, Chapman V (2003) Effects of direct periaqueductal grey administration of a cannabinoid receptor agonist on nociceptive and aversive responses in rats. Neuropharmacology 45:594–604

    Article  CAS  PubMed  Google Scholar 

  • Fogaça MV, Aguiar DC, Moreira FA, Guimarães FS (2012) The endocannabinoid and endovanilloid systems interact in the rat prelimbic medial prefrontal cortex to control anxiety-like behavior. Neuropharmacology 63:202–210

    Article  CAS  PubMed  Google Scholar 

  • Fogaça MV, Gomes FV, Moreira FA, Guimarães FS, Aguiar DC (2013) Effects of glutamate NMDA and TRPV1 receptor antagonists on the biphasic responses to anandamide injected into the dorsolateral periaqueductal grey of Wistar rats. Psychopharmacology 226:579–587

    Article  CAS  PubMed  Google Scholar 

  • Freund TF, Katona I, Piomelli D (2003) Role of endogenous cannabinoids in synaptic signaling. Physiol Rev 83:1017–1066

    Article  CAS  PubMed  Google Scholar 

  • Griebel G, Blanchard DC, Blanchard RJ (1996) Predator-elicited flight responses in Swiss-Webster mice: an experimental model of panic attacks. Prog Neuro-Psychopharmacol Biol Psychiatry 20:185–205

    Article  CAS  Google Scholar 

  • Gross CT, Canteras NS (2012) The many paths to fear. Nat Rev Neurosci 13:651–658

    Article  CAS  PubMed  Google Scholar 

  • Heinz DE, Genewsky A, Wotjak CT (2017) Enhanced anandamide signaling reduces flight behavior elicited by an approaching robo-beetle. Neuropharmacology 126:233–241

    Article  CAS  PubMed  Google Scholar 

  • Lammers JHCM, Kruk MR, Meelis W, van der Poel AM (1988) Hypothalamic substrates for brain stimulation-induced patterns of locomotion and escape jumps in the rat. Brain Res 449:294–310

    Article  CAS  PubMed  Google Scholar 

  • Lisboa SF, Camargo LHA, Magesto AC, Resstel LBM, Guimarães FS (2014) Cannabinoid modulation of predator fear: involvement of the dorsolateral periaqueductal gray. Int J Neuropsychopharmacol 17:1193–1206

    Article  CAS  PubMed  Google Scholar 

  • Lisboa SF, Guimarães FS (2012) Differential role of CB1 and TRPV1 receptors on anandamide modulation of defensive responses induced by nitric oxide in the dorsolateral periaqueductal gray. Neuropharmacology 62:2455–2462

    Article  CAS  PubMed  Google Scholar 

  • Luchicchi A, Pistis M (2012) Anandamide and 2-arachidonoylglycerol: pharmacological properties, functional features, and emerging specificities of the two major endocannabinoids. Mol Neurobiol 46:374–392

    Article  CAS  Google Scholar 

  • Lutz B, Marsicano G, Maldonado R, Hillard CJ (2015) The endocannabinoid system in guarding against fear, anxiety and stress. Nat Rev Neurosci 16:705–718

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mascarenhas DC, Gomes KS, Nunes-de-Souza RL (2013) Anxiogenic-like effect induced by TRPV1 receptor activation within the dorsal periaqueductal gray matter in mice. Behav Brain Res 250:308–315

    Article  CAS  PubMed  Google Scholar 

  • Micale V, Cristino L, Tamburella A, Petrosino S, Leggio GM, Drago F, Di Marzo V (2009) Anxiolytic effects in mice of a dual blocker of fatty acid amide hydrolase and transient receptor potential vanilloid type-1 channels. Neuropsychopharmacology 34:593–606

    Article  CAS  PubMed  Google Scholar 

  • Milani H, Graeff FG (1987) GABA-benzodiazepine modulation of aversion in the medial hypothalamus of the rat. Pharmacol Biochem Behav 28:21–27

    Article  CAS  PubMed  Google Scholar 

  • Mobbs D, Marchant JL, Hassabis D, Seymour B, Tan G, Gray M, Petrovic P, Dolan RJ, Frith CD (2009) From threat to fear: the neural organization of defensive fear systems in humans. J Neurosci 29:12236–12243

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moreira FA, Aguiar DC, Guimarães FS (2007) Anxiolytic-like effect of cannabinoids injected into the rat dorsolateral periaqueductal gray. Neuropharmacology 52:958–965

    Article  CAS  PubMed  Google Scholar 

  • Moreira FA, Aguiar DC, Terzian ALB, Guimarães FS, Wotjak CT (2012) Cannabinoid type 1 receptors and transient receptor potential vanilloid type 1 channels in fear and anxiety-two sides of one coin? Neuroscience 204:186–192

    Article  CAS  PubMed  Google Scholar 

  • Paschoalin-Maurin T, dos Anjos-Garcia T, Falconi-Sobrinho LL, de Freitas RL, Coimbra JPC, Laure CJ, Coimbra NC (2018) The rodent-versus-wild snake paradigm as a model for studying anxiety- and panic-like behaviors: face, construct and predictive validities. Neuroscience 369:336–349

    Article  CAS  PubMed  Google Scholar 

  • Paxinos G, Franklin KBJ (2001) The mouse brain in stereotaxic coordinates. Academic Press, San Diego

    Google Scholar 

  • Rubino T, Realini N, Castiglioni C, Guidali C, Vigano D, Marras E, Petrosino S, Perletti G, Maccarrone M, Di Marzo V, Parolaro D (2008) Role in anxiety behavior of the endocannabinoid system in the prefrontal cortex. Cereb Cortex 18:1292–1301

    Article  CAS  PubMed  Google Scholar 

  • Schoenen J, Di Clemente L, Vandenheede M, Fumal A, De Pasqua V, Mouchamps M, Remacle JM, de Noordhout AM (2005) Hypothalamic stimulation in chronic cluster headache: a pilot study of efficacy and mode of action. Brain 128:940–947

    Article  CAS  PubMed  Google Scholar 

  • Shekhar A (1994) Effects of treatment with imipramine and clonazepam on an animal model of panic disorder. Biol Psychiatry 36:748–758

    Article  CAS  PubMed  Google Scholar 

  • Tóth A, Boczán J, Kedei N, Lizanecz E, Bagi Z, Papp Z, Édes I, Csiba L, Blumberg PM (2005) Expression and distribution of vanilloid receptor 1 (TRPV1) in the adult rat brain. Mol Brain Res 135:162–168

    Article  CAS  PubMed  Google Scholar 

  • Ullah F, dos Anjos-Garcia T, dos Santos IR, Biagioni AF, Coimbra NC (2015) Relevance of dorsomedial hypothalamus, dorsomedial division of the ventromedial hypothalamus and the dorsal periaqueductal gray matter in the organization of freezing or oriented and non-oriented escape emotional behaviors. Behav Brain Res 293:143–152

    Article  PubMed  Google Scholar 

  • Ullah F, dos Anjos-Garcia T, Mendes-Gomes J, Elias-Filho DH, Falconi-Sobrinho LL, de Freitas RL, Khan AU, Oliveira R, Coimbra NC (2017) Connexions between the dorsomedial division of the ventromedial hypothalamus and the dorsal periaqueductal grey matter are critical in the elaboration of hypothalamically mediated panic-like behaviour. Behav Brain Res 319:135–147

    Article  CAS  PubMed  Google Scholar 

  • Uribe-Mariño A, Francisco A, Castiblanco-Urbina MA, Twardowschy A, Salgado-Rohner CJ, Crippa JAS, Hallak JEC, Zuardi AW, Coimbra NC (2012) Anti-aversive effects of cannabidiol on innate fear-induced behaviors evoked by an ethological model of panic attacks based on a prey vs the wild snake Epicrates cenchria crassus confrontation paradigm. Neuropsychopharmacology 37:412–421

    Article  CAS  PubMed  Google Scholar 

  • Viana TG, Hott SC, Resstel LB, Aguiar DC, Moreira FA (2015) Anti-aversive role of the endocannabinoid system in the periaqueductal gray stimulation model of panic attacks in rats. Psychopharmacology 232:1545–1553

    Article  CAS  PubMed  Google Scholar 

  • Wilent WB, Oh MY, Buetefisch CM, Bailes JE, Cantella D, Angle C, Whiting DM (2010) Induction of panic attack by stimulation of the ventromedial hypothalamus. J Neurosurg 112:1295–1298

    Article  PubMed  Google Scholar 

  • Zarrindast MR, Sarahroodi S, Arzi A, Khodayar MJ, Taheri-Shalmani S, Rezayof A (2008) Cannabinoid CB1 receptors of the rat central amygdala mediate anxiety-like behavior: interaction with the opioid system. Behav Pharmacol 19:716–723

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Daoud Hibrahim Elias Filho for expert technical assistance with the immunohistochemistry procedure. D.H. Elias-Filhos was the recipient of technician scholarships from FAPESP (TT-2, grant 02/01497-1) and CNPq (grant 372838/2018-9).

Funding

This work was supported by the Conselho Nacional de Pesquisa e Desenvolvimento Tecnológico (CNPq) (grants 470119/2004-7 and 427397/2018-9)), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (grants 2012/03798-0, 2017/11855-8), and a Pro-Rectory of the University of São Paulo (USP) research grant (NAP-USP-NuPNE; grant IaPq2012-156-USP-12.1.25440.01.6). T. dos Anjos-Garcia was financially supported by CNPq (Scientiae Doctor Fellowship, grant 141124/2014-8) and is a postdoctorate researcher supported by FAPESP (grant 2917/22647-7). None of the organizations played further roles in the study design; the collection, analysis, or interpretation of the data; the writing of the report; or the decision to submit the paper for publication. N.C. Coimbra is a researcher (level 1A) from CNPq (grant 301341/2015-0).

Author information

Authors and Affiliations

  1. Laboratory of Neuroanatomy and Neuropsychobiology, Department of Pharmacology, Ribeirão Preto Medical School of the University of São Paulo (FMRP-USP), Av. Bandeirantes 3900, Ribeirão Preto, São Paulo, 14049-900, Brazil

    Tayllon dos Anjos-Garcia & Norberto Cysne Coimbra

  2. NAP-USP-Neurobiology of Emotions Research Centre (NuPNE), Ribeirão Preto Medical School of the University of São Paulo (FMRP-USP), Av. Bandeirantes, 3900, Ribeirão Preto, São Paulo, 14049-900, Brazil

    Tayllon dos Anjos-Garcia & Norberto Cysne Coimbra

  3. Ophidiarium LNN-FMRP-USP/INeC, Ribeirão Preto School of Medicine of the University of São Paulo (FMRP-USP), Av. Bandeirantes, 3900, Ribeirão Preto, São Paulo, 14049-900, Brazil

    Tayllon dos Anjos-Garcia & Norberto Cysne Coimbra

  4. Behavioural Neuroscience Institute (INeC), Av. do Café, 2450, Ribeirão Preto, São Paulo, 14050-220, Brazil

    Norberto Cysne Coimbra

Authors
  1. Tayllon dos Anjos-Garcia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Norberto Cysne Coimbra
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Norberto Cysne Coimbra.

Ethics declarations

The experimental procedures were approved by the Commission of Ethics in Animal Experimentation (CONCEA process 227/2014) of the Ribeirão Preto Medical School of the University of São Paulo (FMRP-USP).

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

dos Anjos-Garcia, T., Coimbra, N.C. Opposing roles of dorsomedial hypothalamic CB1 and TRPV1 receptors in anandamide signaling during the panic-like response elicited in mice by Brazilian rainbow Boidae snakes. Psychopharmacology 236, 1863–1874 (2019). https://doi.org/10.1007/s00213-019-5170-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00213-019-5170-2

Keywords

Search

Navigation