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Concentration, population, and context-dependent effects of AM251 in zebrafish

Abstract

Rationale

The function of the cannabinoid type 1 receptor (CB1-R) is poorly understood in zebrafish, and numerous inconsistent effects have been reported on it in the literature.

Objective

The objective of the present study is to determine whether differences in the reported effects of CB1-R antagonism on anxiety-like behavioural responses, dopaminergic and serotonergic responses are due to concentration, context-dependent and/or population (genotype-related) effects.

Method

Two genetically distinct populations of zebrafish (AB and short fin (SF)) were treated with different concentrations of AM251 (0, 0.1, 1mg/L), and behavioural responses were quantified under two different contexts: one, following habituation and two, subsequently in a novel environment. The levels of dopamine, serotonin and their metabolites 3,4-dihydroxyindole acetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA) were quantified from whole-brain tissue.

Results

We demonstrate that a 60-min exposure to AM251 (0, 0.1, 1mg/L) does not alter behavioural performance following habituation in either populations. However, when subsequently transferred to a novel environment, zebrafish that were pre-treated with the highest dose of AM251 (1mg/L) exhibited increased anxiety-like behavioural responses including elevated absolute turn angle, freezing and bottom dwelling. We found that exposure to the highest dose of AM251 (1mg/L) for 60min increased serotonin in fish of both populations tested. In contrast, exposure to 0.1mg/L AM251 decreased, whereas to 1mg/L AM251 increased dopamine, DOPAC and 5-HIAA in fish of both populations.

Conclusion

Our results demonstrate a genotype-independent effect of AM251 but imply that the inconsistent findings obtained after pharmacological blockade of CB1-Rs in zebrafish may be due to a combination of concentration- and environmental context-dependent effects.

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References

  • Akhtar MT, Ali S, Rashidi H, van der Kooy F, Verpoorte R, Richardson MK (2013) Developmental effects of cannabinoids on zebrafish larvae. Zebrafish 10:283–293

    CAS  Article  PubMed  Google Scholar 

  • Barba-Escobedo PA, Gould GG (2012) Visual social preferences of lone zebrafish in a novel environment; strain and anxiolytic effects. Genes Brain Behav 11:366–373

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Blaser RE, Chadwick L, McGinnis GC (2010a) Behavioural measures of anxiety in zebrafish (Danio rerio). Behav Brain Res 208:56–62

    CAS  Article  PubMed  Google Scholar 

  • Blaser RE, Koid A, Poliner RM (2010b) Context-dependent sensitization to ethanol in zebrafish (Danio rerio). Pharmacol Biochem Behav 95:278–284

    CAS  Article  PubMed  Google Scholar 

  • Bortolotto JW, Cognato GP, Christoff RR, Roesler LN, Leite CE, Kist LW, Bogo MR, Vianna MR, Bonan CD (2014) Long-term exposure to paraquat alters behavioural parameters and dopamine levels in adult zebrafish (Danio rerio). Zebrafish 11:142–153

    CAS  Article  PubMed  Google Scholar 

  • Braida D, Limonta V, Pegorini S, Zani A, Guerini-Rocco C, Gori E, Sala M (2007) Hallucinatory and rewarding effect of salvinorin a in zebrafish: kappa-opioid and CB1-cannabinoid receptor involvement. Psychopharmacol 190:441–448

    CAS  Article  Google Scholar 

  • Chatterjee D, Gerlai R (2009) High precision liquid chromatography analysis of dopaminergic and serotonergic responses to acute alcohol exposure in zebrafish. Behav Brain Res 200:208–213

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Chatterjee D, Shams S, Gerlai R (2014) Chronic and acute alcohol administration administration induced neurochemical changes in the brain: comparison of distinct zebrafish populations. Amino Acids 46:921–930

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Connors KA, Valenti TW, Lawless K, Sackerman J, Onaivi ES, Brooks BW, Gould GG (2014) Similar anxiolytic effects of agonists targeting serotonin 5-HT1A or cannabinoid CB receptors on zebrafish behaviour in novel environments. Aquat Toxicol 151:105–113

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Dazzi L, Talani G, Biggio F, Utzeri C, Lallai V, Licheri V, Lutzu S, Mostallino MC, Secci PP, Sanna E (2014) Involvement of the cannabinoid CB1 receptor in modulation of dopamine output in the prefrontal cortex associated with food restriction in rats. PLoS One 9:e92224

    Article  PubMed  PubMed Central  Google Scholar 

  • Gerlai R, Chatterjee D, Pereira T, Sawashima T, Krishnannair R (2009) Acute and chronic alcohol dose: population differences in behaviour and neurochemistry of zebrafish. Genes Brain Behav 8:586–599

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Guryev V (2006) Genetic variation in the zebrafish. Genome Res 4:491–497

    Article  Google Scholar 

  • Haring M, Kaiser N, Monory K, Lutz B (2011) Circuit specific functions on cannabinoid Cb1 receptor in the balance of investigatory drive and exploration. PLoS One 6:e26617

    Article  PubMed  PubMed Central  Google Scholar 

  • Haring M, Grieb M, Monory K, Lutz B, Moreira F (2013) Cannabinoid CB1 receptor in the modulation of stress coping behaviour in mice: the role of serotonin and different forebrain neuronal subpopulations. Neuropharmacol 65:83–89

    CAS  Article  Google Scholar 

  • Haring M, Enk V, Aparisi Rey A, Loch S, Ruiz de Azua I, Weber T, Bartsch D, Monory K, Lutz B (2015) Cannabinoid type-1 receptor signalling in central serotonergic neurons regulates anxiety-like behaviour and sociability. Front Behav Neurosci 9:235

    Article  PubMed  PubMed Central  Google Scholar 

  • Huang J, Zhong Z, Wang M, Chen X, Tan Y, Zhang S, He W, He X, Huang G, Lu H, Wu P, Che Y, YanYL PJH, Chen W, Wang H (2015) Circadian modulation of dopamine levels and dopaminergic neuron development contributes to attention deficiency and hyperactive behaviour. J Neurosci 35:2572–2587

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Jay M, De Faveri F, McDearmid JR (2015) Firing dynamics and modulatory actions of supraspinal dopaminergic neurons during zebrafish locomotor behaviour. Curr Biol 25:435–444

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Lafenetre P, Chaouloff F, Marsicano G (2009) Bidirectional regulation of novelty-induced behavioural inhibition by the endocannabinoid system. Neuropharmacol 57:715–721

    CAS  Article  Google Scholar 

  • Lam CS, Rastegar S, Strahle U (2006) Distribution of cannabinoid receptor 1 in the CNS of zebrafish. Neuroscience 138:83–95

  • Levin ED, Bencan Z, Cerutti DT (2007) Anxiolytic effects of nicotine in zebrafish. Physiol Behav 90:54–58

    CAS  Article  PubMed  Google Scholar 

  • Liu X, Guo N, Lin J, Zhang Y, Chen XQ, Li S, He L, Li Q (2014) Strain-dependent differential behavioural responses of zebrafish larvae to acute MK-801 treatment. Pharmacol Biochem Behav 127:82–89

    CAS  Article  PubMed  Google Scholar 

  • Marsicano G, Lutz B (1999) Expression of the cannabinoid receptor CB1 in distinct neuronal subpopulations in the adult mouse forebrain. Eur J Neurosci 11:4213–4225

    CAS  Article  PubMed  Google Scholar 

  • Maximino C, Puty B, Benzecry R, Araujo J, Lima MG, de Jesus Oliveira Batista E, Renata de Matos Oliveira K, Crespo-Lopez ME, Herculano AM (2013) Role of serotonin in zebrafish (Danio rerio) anxiety: relationship with serotonin levels and effect of buspirone, WAY 100635, SB 224289, fluoxetine and para-chlorophenylalanine (pCPA) in two behavioural models. Neuropharmacol 71:83–97

    CAS  Article  Google Scholar 

  • Maximino C, Lima MG, Costa CC, Guedes IM, Herculano AM (2014) Fluoxetine and WAY 100,635 dissociate increases in scototaxis and analgesia induced by alarm substance in zebrafish (Danio rerio Hamilton 1822). Pharmacol Biochem Behav 124:425–433

    CAS  Article  PubMed  Google Scholar 

  • McPartland JM, Glass M, Matias I, Norris RW, Kilpatrick CW (2007) A shifted repertoire of endocannabinoid genes in the zebrafish (Danio rerio). Mol Genet Genomics 277:555–570

    CAS  Article  PubMed  Google Scholar 

  • Migliarini B, Carnevali O (2008) Anandamide modulates growth and lipid metabolism in the zebrafish Danio rerio. Mol Cell Endocrinol 286S:12–16

    Article  Google Scholar 

  • Migliarini B, Carnevali O (2009) A novel role for the endocannabinoid system during zebrafish development. Mol Cell Endocrinol 299:172–177

    CAS  Article  PubMed  Google Scholar 

  • Nowicki M, Tran S, Muraleetharan A, Markovic S, Gerlai R (2014) Serotonin antagonist induce anxiolytic and anxiogenic-like behaviour in zebrafish in a receptor-subtype dependent manner. Pharmacol Biochem Behav 126:170–180

    CAS  Article  PubMed  Google Scholar 

  • Pan Y, Chatterjee D, Gerlai R (2012) Strain dependent gene expression and neurochemical levels in the brain of zebrafish: focus on a few alcohol related targets. Physiol Behav 107:773–780

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Pannia E, Tran S, Rampersad M, Gerlai R (2014) Acute ethanol exposure induces behavioural differences in two zebrafish (Danio rerio) strains: a time course analysis. Behav Brain Res 259:174–185

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Pertwee R (2007) GPR55: a new member of the cannabinoid receptor clan? Br J Pharmacol 152:984–986

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Pertwee RG (2008) Ligands that target cannabinoid receptors in the brain: from THC to anandamide and beyond. Addict Biol 13:147–159

    CAS  Article  PubMed  Google Scholar 

  • Quadros VA, Silveira A, Giuliani GS, Didonet F, Silveira AS, Nunes ME, Silva TO, Loro VL, Rosemberg DB (2016) Strain- and context-dependent behavioural responses of acute alarm substance exposure in zebrafish. Behav Process 122:1–11

    Article  Google Scholar 

  • Rey AA, Purrio M, Viveros MP, Lutz B (2012) Biphasic effects of cannabinoids in anxiety responses: CB1 and GABA(B) receptors in the balance of GABAergic and glutamatergic neurotransmission. Neuropsychopharmacology 37:2624–2634

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Ruhl T, Prinz N, Oellers N, Seidel NI, Jonas A, Albayram O, Bilkei-Gorzo A, von der Emde G (2014) Acute administration of THC impairs spatial but not associative memory function in zebrafish. Psychopharmacol 231:3829–3842

    CAS  Article  Google Scholar 

  • Ruhl T, Moesbauer K, Oellers N, von der Emde G (2015) The endocannabinoid system and associative learning and memory in zebrafish. Behav Brain Res 290:61–69

    CAS  Article  PubMed  Google Scholar 

  • Ryberg E, Larsson N, Sjogren S, Hiorth S, Hermansson NO, Leonova J, Elebring T, Nilsson K, Drmota T, Greasley PJ (2007) The orphan receptor GPR55 is a novel cannabinoid receptor. Br J Pharmacol 152:1092–1101

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Sackerman J, Donegan JJ, Cunningham CS, Nguyen NN, Lawless K, Long A, Benno RH, Gould GG (2010) Zebrafish behavior in novel environments: effects of acute exposure to anxiolytic compounds and choice of Danio rerio line. Int J Comp Psychol 23:43–61

    PubMed  PubMed Central  Google Scholar 

  • Simone JJ, Green MR, Hodges TE, McCormick CM (2015a) Differential effects of CB1 receptor antagonism in behavioural tests of unconditioned and conditioned fear in adult male rats. Behav Brain Res 279:9–16

    CAS  Article  PubMed  Google Scholar 

  • Simone JJ, Malivoire BL, McCormick CM (2015b) Effects of CB1 receptor agonism and antagonism on behavioural fear and physiological stress responses in adult intact, ovariectomized, and estradiol-replaced female rats. Neuroscience 306:123–137

    CAS  Article  PubMed  Google Scholar 

  • Song J, Ampatzis K, Ausborn J, El Manira A (2015) A hardwired circuit supplemented with endocannabinoids encodes behavioural choice in zebrafish. Curr Biol 25:2610–2620

    CAS  Article  PubMed  Google Scholar 

  • Stewart AM, Kalueff AV (2014) The behavioural effects of acute delta(9)-tetrahydrocannabinol and heroin (diacetylmorphine) exposure in adult zebrafish. Brain Res 1543:109–119

    CAS  Article  PubMed  Google Scholar 

  • Thiemann G, Watt CA, Ledent C, Molleman A, Hasenohrl RU (2009) Modulation of anxiety by acute blockade and genetic deletion of the CB(1) cannabinoid receptor in mice together with biogenic amine changes in the forebrain. Behav Brain Res 200:60–67

    CAS  Article  PubMed  Google Scholar 

  • Tran S, Gerlai R (2013) Time-course of behavioural changes induced by ethanol in zebrafish (Danio rerio). Behav Brain Res 252:204–213

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Tran S, Chatterjee D, Gerlai R (2014) Acute net stressor increases whole-body cortisol levels without altering whole-brain monoamines in zebrafish. Behav Neurosci 128:621–624

    Article  PubMed  Google Scholar 

  • Tran S, Nowicki M, Muraleetharan A, Gerlai R (2015a) Differential effects of dopamine D1 and D2/3 receptor antagonism on motor responses. Pyschopharmacol 232:795–806

    CAS  Article  Google Scholar 

  • Tran S, Chatterjee D, Gerlai R (2015b) An integrative analysis of ethanol and withdrawal in zebrafish (Danio rerio). Behav Brain Res 276:161–170

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Tran S, Muraleetharan A, Fulcher N, Chatterjee D, Gerlai R (2015c) MK-801 increases locomotor activity in a context-dependent manner in zebrafish. Behav Brain Res 296:26–29

    Article  PubMed  Google Scholar 

  • Tran S, Nowicki M, Muraleetharan A, Chatterjee D, Gerlai R (2015d) Neurochemical factors underlying individual differences in locomotor activity and anxiety-like behavioural responses in zebrafish. Prog Neuropsychopharmacol Biol Psychiatry 65:25–33

    Article  PubMed  Google Scholar 

  • Tran S, Nowicki M, Muraleetharan A, Chatterjee D, Gerlai R (2015e) Differential effects of acute administration of SCH-23390, a D1 receptor antagonist, and of ethanol on swimming activity, anxiety-related responses, and neurochemistry of zebrafish. Psychopharmacol 232:3709–3718

    CAS  Article  Google Scholar 

  • Viveros MP, Marco EM, File SE (2005) Endocannabinoid system and stress and anxiety responses. Pharmacol Biochem Behav 81:331–342

    CAS  Article  PubMed  Google Scholar 

  • Wahlsten D (1990) Insensitivity of the analysis of variance of heredity-environment interaction. Behav Brain Sci 13:109–120

    Article  Google Scholar 

  • Wong K, Elegante M, Bartels B, Elkhayat S, Tien D, Roy S, Goodspeed J, Suciu C, Tan J, Grimes C, Chung A, Rosemberg M, Gaikwad S, Denmark A, Jackson A, Kadri F, Chung KM, Stewart A, Gilder T, Beeson E, Zapolsky I, Wu N, Cachat J, Kalueff AV (2010) Analyzing habituation responses to novelty in zebrafish (Danio rerio). Behav Brain Res 208:450–457

    CAS  Article  PubMed  Google Scholar 

  • Zarrindast MR, Mahboobi S, Sadat-Shirazi MS, Ahmadi S (2011) Anxiolytic effect induced by the cannabinoid CB1 receptor agonist, arachydonilcyclopropylamide (ACPA), in the rat amygdala is mediated through the D1 and D2 dopaminergic systems. J Psychopharmacol 25:131–140

    CAS  Article  PubMed  Google Scholar 

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Acknowledgments

The research was supported by an NSERC Discovery Grant (#311637) and a Brain Canada Foundation Grant issued to Robert Gerlai and an NSERC Canadian Graduate Scholarship (doctoral level) issued to Steven Tran.

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Correspondence to Steven Tran or Robert Gerlai.

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The research conducted was reviewed and approved by the Local Animal Care Committee (LACC) at the University of Toronto Mississauga and is in accordance with the guidelines created by the Canadian Council for Animal Care (CCAC).

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Tran, S., Chatterjee, D., Facciol, A. et al. Concentration, population, and context-dependent effects of AM251 in zebrafish. Psychopharmacology 233, 1445–1454 (2016). https://doi.org/10.1007/s00213-016-4240-y

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  • DOI: https://doi.org/10.1007/s00213-016-4240-y

Keywords

  • CB1 receptor
  • AM251
  • Anxiety
  • Dopamine
  • Serotonin