Skip to main content

Diazepam blocks 50 kHz ultrasonic vocalizations and stereotypies but not the increase in locomotor activity induced in rats by amphetamine

Abstract

Rationale

We have recently shown that the benzodiazepine diazepam inhibits dopamine release in the NAc and blocks the increased release of dopamine induced by DL-amphetamine. Rewarding stimuli and many drugs of abuse can induce dopamine release in the nucleus accumbens as well as 50-kHz ultrasonic vocalizations (USVs) in rats.

Objectives

In the present study, we tested the hypothesis that diazepam can also block the increase in locomotor activity and USVs elicited by amphetamine.

Methods

Fifty-kilohertz USVs, stereotypy, and locomotor behavior were scored in adult male Wistar rats treated with i.p. injections of saline, 3 mg/kg DL-amphetamine, 2 mg/kg diazepam, 0.2 mg/kg haloperidol, or a combination of these drugs.

Results

In agreement with previous studies, amphetamine caused significant increases in the number of USV calls, stereotypies, and locomotor activity. The D2 dopamine receptor antagonist haloperidol blocked the effects of amphetamine on USVs, stereotypy, and locomotor activity. Diazepam blocked the effect of amphetamine on USV and stereotypy, but not on horizontal locomotion.

Conclusions

These results suggest that diazepam blocks the rewarding effect of amphetamine. This finding is promising for basic research regarding treatments of substance use disorders and evaluation of the impact of benzodiazepines on motivation.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  • Ahrens AM, Ma ST, Maier EY, Duvauchelle CL, Schallert T (2009) Repeated intravenous amphetamine exposure: rapid and persistent sensitization of 50-kHz ultrasonic trill calls in rats. Behav Brain Res 197:205–209

    Article  PubMed  CAS  Google Scholar 

  • Ahrens AM, Nobile CW, Page LE, Maier EY, Duvauchelle CL, Schallert T (2013) Individual differences in the conditioned and unconditioned rat 50-kHz ultrasonic vocalizations elicited by repeated amphetamine exposure. Psychopharmacology 229:687–700

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Angrist B, Lee HK, Gershon S (1974) Antagonism of amphetamine-induced symptomatology by a neuroleptic. Am J Psychiatr 131:817–819

    Article  PubMed  CAS  Google Scholar 

  • Barker DJ, Root DH, Ma S, Jha S, Megehee L, Pawlak AP, West MO (2010) Dose-dependent differences in short ultrasonic vocalizations emitted by rats during cocaine self-administration. Psychopharmacology 211:435–442

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Barker DJ, Simmons SJ, Servilio LC, Bercovicz D, Ma S, Root DH, Pawlak AP, West MO (2014) Ultrasonic vocalizations: evidence for an affective opponent process during cocaine self administration. Psychopharmacology 231:909–918

    Article  PubMed  CAS  Google Scholar 

  • Barker DJ, Simmons SJ, West MO (2015) Ultrasonic vocalizations as a measure of affect in preclinical models of drug abuse: a review of current findings. Curr Neuropharmacol 13:193–210

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Brenes J, Schwarting RKW (2014) Attribution and expression of incentive salience are differentially signalled by ultrasonic vocalizations in rats. PLoS One 9(7):e102414

    Article  PubMed  PubMed Central  Google Scholar 

  • Brudzynski SM (2005) Principles of rat communication: quantitative parameters of ultrasonic calls in rats. Behav Genet 35:85–92

    Article  PubMed  Google Scholar 

  • Brudzynski SM (2013) Ethotransmission: communication of emotional states through ultrasonic vocalization in rats. Curr Opin Neurobiol 23:310–317

    Article  PubMed  CAS  Google Scholar 

  • Brudzynski SM (2015) Pharmacology of ultrasonic vocalizations in adult rats: significance, call classification and neural substrate. Curr Neuropharmacol 13:180–192

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Burgdorf J, Knutson B, Panksepp J, Ikemoto S (2001) Nucleus accumbens amphetamine microinjections unconditionally elicit 50-kHz ultrasonic vocalizations in rats. Behav Neurosci 115:940–944

    Article  PubMed  CAS  Google Scholar 

  • Burgdorf J, Kroes RA, Moskal JR, Pfaus JG, Brudzynski SM, Panksepp J (2008) Ultrasonic vocalizations of rats (Rattus norvegicus) during mating, play, and aggression: behavioral concomitants, relationship to reward, and self-administration of playback. J Comp Psychol 122:357–367

    Article  PubMed  Google Scholar 

  • Ciucci MR, Ma ST, Fox C, Kane JR, Ramig LO, Schallert T (2007) Qualitative changes in ultrasonic vocalization in rats after unilateral dopamine depletion or haloperidol: a preliminary study. Behav Brain Res 182:284–289

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Clark D, Furmidge LJ, Petry N, Tong ZY, Ericsson M, Johnson D (1991) Behavioral profile of partial D2 dopamine receptor agonists. 1. Atypical inhibition of D-amphetamine-induced locomotor hyperactivity and stereotypy. Psychopharmacology 105:381–392

    Article  PubMed  CAS  Google Scholar 

  • Clark JJ, Sandberg SG, Wanat MJ, Gan JO, Horne EA, Hart AS, Akers CA, Parker JG, Willuhn I, Martinez V, Evans SB, Stella N, Phillips PE (2010) Chronic microsensors for longitudinal, subsecond dopamine detection in behaving animals. Nat Methods 7:126–129

    Article  PubMed  CAS  Google Scholar 

  • Creese I, Iversen SD (1973) Blockage of amphetamine induced motor stimulation and stereotypy in adult rat following neonatal treatment with 6-hydroxydopamine. Brain Res 55:369–382

    Article  PubMed  CAS  Google Scholar 

  • Di Chiara G, Imperato A (1988) Drugs abused by humans preferentially increase synaptic dopamine concentrations in the mesolimbic system of freely moving rats. Proc Natl Acad Sci U S A 85:5274–5278

    Article  PubMed  PubMed Central  Google Scholar 

  • Easton N, Steward C, Marshal F, Fone K, Marsden C (2007) Effects of amphetamine isomers, methylphenidate and atomoxetine on synaptosomal and synaptic vesicle accumulation and release of dopamine and noradrenaline in vitro in the rat brain. Neuropharmacology 52:405–414

    Article  PubMed  CAS  Google Scholar 

  • Eilam D, Clements KVA, Szechtman H (1991) Differential-effects of d1-dopamine and D2-dopamine agonists on stereotyped locomotion in rats. Behav Brain Res 45:117–124

    Article  PubMed  CAS  Google Scholar 

  • Finlay JM, Damsma G, Fibiger HC (1992) Benzodiazepine-induced decreases in extracellular concentrations of dopamine in the nucleus-accumbens after acute and repeated administration. Psychopharmacology 106:202–208

    Article  PubMed  CAS  Google Scholar 

  • Flagel SB, Clark JJ, Robinson TE, Mayo L, Czuj A, Willuhn I, Akers CA, Clinton SM, Phillips PE, Akil H (2011) A selective role for dopamine in stimulus-reward learning. Nature 469:53–57

    Article  PubMed  CAS  Google Scholar 

  • Fleckenstein AE, Volz TJ, Riddle EL, Gibb JW, Hanson GR (2007) New insights into the mechanism of action of amphetamines. Annu Rev Pharmacol Toxicol 47:681–698

    Article  PubMed  CAS  Google Scholar 

  • Goeders NE (1997) A neuroendocrine role in cocaine reinforcement. Psychoneuroendocrinology 22:237–259

    Article  PubMed  CAS  Google Scholar 

  • Gomez-A A, Fiorenza AM, Boschen SL, Sugi AH, Beckman D, Ferreira ST, Lee K, Blaha CH, Da Cunha C (2017) Diazepam inhibits electrically evoked and tonic dopamine release in the nucleus accumbens and reverses the effect of amphetamine. A C S Chem Neurosci 8:300–309

    Article  CAS  Google Scholar 

  • Hodgson RA, Guthrie DH, Varty GB (2008) Duration of ultrasonic vocalizations in the isolated rat pup as a behavioral measure: sensitivity to anxiolytic and antidepressant drugs. Pharmacol Biochem Behav 88:341–348

    Article  PubMed  CAS  Google Scholar 

  • Ikemoto S (2007) Dopamine reward circuitry: two projection systems from the ventral midbrain to the nucleus accumbens-olfactory tubercle complex. Brain Res Rev 56:27–78

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Imperato A, Di Chiara G (1986) Preferential stimulation of dopamine release in the nucleus accumbens of freely moving rats by ethanol. J Pharmacol Exp Ther 239:219–228

    PubMed  CAS  Google Scholar 

  • Kablinger AS, Lindner MA, Casso S, Hefti F, DeMuth G, Fox BS, McNair LA, McCarthy BG, Goeders NE (2012) Effects of the combination of metyrapone and oxazepam on cocaine craving and cocaine taking: a double-blind, randomized, placebo-controlled pilot study. J Psychopharmacol 26:973–981

    Article  PubMed  CAS  Google Scholar 

  • Knutson B, Burgdorf J, Panksepp J (1999) High-frequency ultrasonic vocalizations index conditioned pharmacological reward in rats. Physiol Behav 66:639–643

    Article  PubMed  CAS  Google Scholar 

  • Koob GF, Volkow ND (2010) Neurocircuitry of addiction. Neuropsychopharmacology 35:217–238

    Article  PubMed  Google Scholar 

  • Kuczenski R, Segal DS, Aizenstein ML (1991) Amphetamine, cocaine, and fencamfamine -relationship between locmotor and stereotypy response profiles and caudate and accumbens-dopamine dynamics. J Neurosci 11:2703–2712

  • Leri F, Franklin KB (2000) Effects of diazepam on conditioned place preference induced by morphine or amphetamine in the rat. Psychopharmacology 150:351–360

    Article  PubMed  CAS  Google Scholar 

  • Ma ST, Maier EY, Ahrens AM, Schallert T, Duvauchelle CL (2010) Repeated intravenous cocaine experience: development and escalation of pre-drug anticipatory 50-kHz ultrasonic vocalizations in rats. Behav Brain Res 212:109–114

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Mackey WB, Vanderkooy D (1985) Neuroleptics block the positive reinforcing effects of amphetamine but not of morphine as measured by place conditioning. Pharmacol Biochem Behav 22:101–105

    Article  PubMed  CAS  Google Scholar 

  • Maier EY, Ahrens AM, Ma ST, Schallert T, Duvauchelle CL (2010) Cocaine deprivation effect: cue abstinence over weekends boosts anticipatory 50-kHz ultrasonic vocalizations in rats. Behav Brain Res 214:75–79

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Maier EY, Abdalla M, Ahrens AM, Schallert T, Duvauchelle CL (2012) The missing variable: ultrasonic vocalizations reveal hidden sensitization and tolerance-like effects during long-term cocaine administration. Psychopharmacology 219:1141–1152

    Article  PubMed  CAS  Google Scholar 

  • Meririnne E, Kankaanpaa A, Lillsunde P, Seppala T (1999) The effects of diazepam and zolpidem on cocaine- and amphetamine-induced place preference. Pharmacol Biochem Behav 62:159–164

    Article  PubMed  CAS  Google Scholar 

  • Mithani S, Martin-Iverson MT, Phillips AG, Fibiger HC (1986) The effects of haloperidol on amphetamine- and methylphenidate induced conditioned place preferences and locomotor activity. Psychopharmacology 90:247–252

    Article  PubMed  CAS  Google Scholar 

  • Mu P, Fuchs T, Saal DB, Sorg BA, Dong Y, Panksepp J (2009) Repeated cocaine exposure induces sensitization of ultrasonic vocalization in rats. Neurosci Lett 453:31–35

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Murai T, Koshikawa N, Kanayama T, Takada K, Tomiyama K, Kobayashi M (1994) Local-administration of flurazepam has different effects on dopamine release in striatum and nucleus accumbens—a microdialysis study. Eur J Pharmacol 261:65–71

    Article  PubMed  CAS  Google Scholar 

  • Natusch C, Schwarting RKW (2010) Using bedding in a test environment critically affects 50 kHz ultrasonic vocalizations in laboratory rats. Pharmacol Biochem Behav 96:251–259

    Article  PubMed  CAS  Google Scholar 

  • O'Brien DP, White FJ (1987) Inhibition of non-dopamine cells in the ventral tegmental area by benzodiazepines—relationship to A10 dopamine cell-activity. Eur J Pharmacol 142:343–354

    Article  PubMed  CAS  Google Scholar 

  • Olivier B, Molewijk E, van Oorschot R, van der Heyden J, Ronken E, Mos J (1998) Rat pup ultrasonic vocalization: effects of benzodiazepine receptor ligands. Eur J Pharmacol 358:117–128

    Article  PubMed  CAS  Google Scholar 

  • Pereira M, Andreatini R, Schwarting RK, Brenes JC (2014) Amphetamine-induced appetitive 50-kHz calls in rats: a marker of affect in mania? Psychopharmacology 231:2567–2577

    Article  PubMed  CAS  Google Scholar 

  • Ringel LE, Basken JN, Grant LM, Ciucci MR (2013) Dopamine D1 and D2 receptor antagonism effects on rat ultrasonic vocalizations. Behav Brain Res 252-259

  • Rippberger H, van Gaalen MM, Schwarting RKW, Wöhr M (2015) Environmental and pharmacological modulation of amphetamine induced 50-kHz ultrasonic vocalizations in rats. Curr Neuropharmacol 13:220–232

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Robertson SD, Matthies HJG, Galli A (2009) A closer look at amphetamine-induced reverse transport and trafficking of the dopamine and norepinephrine transporters. Mol Neurobiol 39:73–80

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Robinson TE, Becker JB (1986) Enduring changes in brain and behavior produced by chronic amphetamine administration: a review and evaluation of animal models of amphetamine psychosis. Brain Res 396:157–198

    Article  PubMed  CAS  Google Scholar 

  • Rowlett JK, Tornatzky W, Cook JM, Ma C, Miczek KA (2001) Zolpidem, triazolam, and diazepam decrease distress vocalizations in mouse pups: differential antagonism by flumazenil and betacarboline-3-carboxylate-t-butyl ester (beta-CCt). J Pharmacol Exp Ther 297:247–253

    PubMed  CAS  Google Scholar 

  • Rudolph U, Knoflach F (2011) Beyond classical benzodiazepines: novel therapeutic potential of GABA(a) receptor subtypes. Nat Rev Drug Discov 10:685–697

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Rush CR, Stoops WW, Wagner FP, Hays LR, Glaser PEA (2004) Alprazolam attenuates the behavioral effects of D-amphetamine in humans. J Clin Psychopharmacol 24:410–420

    Article  PubMed  CAS  Google Scholar 

  • Sadananda M, Natusch C, Karrenbauer B, Schwarting RKW (2012) 50-kHz calls in rats: effects of MDMA and the 5-HT(1A) receptor agonist 8-OH-DPAT. Pharmacol Biochem Behav 101:258–264

    Article  PubMed  CAS  Google Scholar 

  • Sams-Dodd F (1998) Effects of continuous D-amphetamine and phencyclidine administration on social behaviour, stereotyped behaviour, and locomotor activity in rats. Neuropsychopharmacol 19:18–25

  • Schwarting RKW, Wöhr M (2012) On the relationships between ultrasonic calling and anxiety-related behavior in rats. Braz J Med Biol Res 45:337–348

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Schwarting RKW, Natusch C, Llano L, Bedenk B, Borchers M, Wohr M (2007a) 50-kHz ultrasonic calling in male rats: environmental, social and subjective factors. Behav Pharmacol 18:S66–S67

    Google Scholar 

  • Schwarting RKW, Jegan N, Wöhr M (2007b) Situational factors, conditions and individual variables which can determine ultrasonic vocalizations in male adult Wistar rats. Behav Brain Res 182:208–222

    Article  PubMed  Google Scholar 

  • Simola N (2015) Rat Ultrasonic Vocalizations and Behavioral Neuropharmacology: From the Screening of Drugs to the Study of Disease. Current Neuropharmacology 13:164–179

  • Simola N, Ma ST, Schallert T (2010) Influence of acute caffeine on 50-kHz ultrasonic vocalizations in male adult rats and relevance to caffeine-mediated psychopharmacological effects. Int J Neuropsychopharmacol 13:123–132

    Article  PubMed  Google Scholar 

  • Simola N, Fenu S, Costa G, Pinna A, Plumitallo A, Morelli M (2012) Pharmacological characterization of 50-kHz ultrasonic vocalizations in rats: comparison of the effects of different psychoactive drugs and relevance in drug-induced reward. Neuropharmacology 63:224–234

    Article  PubMed  CAS  Google Scholar 

  • Simola N, Frau L, Plumitallo A, Morelli M (2014) Direct and long lasting effects elicited by repeated drug administration on 50-kHz ultrasonic vocalizations are regulated differently: implications for the study of the affective properties of drugs of abuse. Int J Neuropsychopharmacol 17:429–441

    Article  PubMed  CAS  Google Scholar 

  • Spence AL, Guerin GF, Goeders NE (2016) The differential effects of alprazolam and oxazepam on methamphetamine self-administration in rats. Drug Alcohol Depend 166:209–217

    Article  PubMed  CAS  Google Scholar 

  • Straub CJ, WAJr C, Rudolph U (2010) Diazepam and cocaine potentiate brain stimulation reward in C57BL/6J mice. Behav Brain Res 206:17–20

    Article  PubMed  CAS  Google Scholar 

  • Tan KR, Brown M, Labouebe G, Yvon C, Creton C, Fritschy JM, Rudolph U, Luscher C (2010) Neural bases for addictive properties of benzodiazepines. Nature 463:769–U78

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Thompson B, Leonard KC, Brudzynski SM (2006) Amphetamine induced 50 kHz calls from rat nucleus accumbens: a quantitative mapping study and acoustic analysis. Behav Brain Res 168:64–73

    Article  PubMed  CAS  Google Scholar 

  • Volkow ND, Koob GF, McLellan AT (2016) Neurobiologic advances from the brain disease model of addiction. N Engl J Med 374:363–371

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Wendler E, De Souza CP, Vecchia DD, Kanazawa LKS, Soares d A, Hocayen P, Wöhr M, Schwarting RKW, Andreatini R (2016) Evaluation of 50-kHz ultrasonic vocalizations in animal models of mania: ketamine and lisdexamfetamine-induced hyperlocomotion in rats. Eur Neuropsychopharmacol 26:1900–1908

    Article  PubMed  CAS  Google Scholar 

  • Williams SN, Undieh AS (2010) Brain-derived neurotrophic factor signaling modulates cocaine induction of reward-associated ultrasonic vocalization in rats. J Pharmacol Exp Ther 332:463–468

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Willuhn I, Burgeno LM, Everitt BJ, Phillips PE (2012) Hierarchical recruitment of phasic dopamine signaling in the striatum during the progression of cocaine use. Proc Natl Acad Sci U S A 109:20703–20708

    Article  PubMed  PubMed Central  Google Scholar 

  • Wintink AJ, Brudzynski SM (2001) The related roles of dopamine and glutamate in the initiation of 50-kHz ultrasonic calls in adult rats. Pharmacol Biochem Behav 70:317–323

    Article  PubMed  CAS  Google Scholar 

  • Wise RA, Rompre PP (1989) Brain dopamine and reward. Annu Rev Psychol 40:191–225

    Article  PubMed  CAS  Google Scholar 

  • Wöhr M, Houx B, Schwarting RKW, Spruijt B (2008) Effects of experience and context on 50-kHz vocalizations in rats. Physiol Behav 93:766–776

    Article  PubMed  CAS  Google Scholar 

  • Wolgin DL (2012) Amphetamine stereotypy, the basal ganglia, and the “selection problem”. Behav Brain Res 231:297–308

    Article  PubMed  CAS  Google Scholar 

  • Woods JH, Katz JL, Winger G (1992) Benzodiazepines—use, abuse, and consequences. Pharmacol Rev 44:151–347

    PubMed  CAS  Google Scholar 

  • Wright JM, Gourdon JC, Clarke PB (2010) Identification of multiple call categories within the rich repertoire of adult rat 50-kHz ultrasonic vocalizations: effects of amphetamine and social context. Psychopharmacology 211:1–13

    Article  PubMed  CAS  Google Scholar 

  • Wright JM, Dobosiewicz MR, Clarke PB (2013) The role of dopaminergic transmission through D1-like and D2-like receptors in amphetamine induced rat ultrasonic vocalizations. Psychopharmacology 225:853–868

    Article  PubMed  CAS  Google Scholar 

  • Yoshida Y, Koide S, Hirose N, Takada K, Saigusa T, Koshikawa N (1999) In vivo microdialysis evidence that midazolam facilitates propofol-induced reduction in rat accumbal dopamine release. Neurosci Res Commun 25:121–127

    Article  CAS  Google Scholar 

  • Zetterstrom T, Fillenz M (1990) Local-administration of flurazepam has different effects on dopamine release in striatum and nucleus accumbens—a microdialysis study. Neuropharmacology 29:129–134

    Article  PubMed  CAS  Google Scholar 

Download references

Contributors

C.D.C, R.A., D.L.R., and R.K.W.S. designed the research; G.O.G. and D.D.V performed the research; G.O.G., C.D.C, D.D.V. D.L.R., and R.A. analyzed the data; and G.O.G. and C.D.C wrote the paper.

Role of the funding source

GOG, DDV, RA, and CDC were supported by INNT, CNPq, CAPES, and UFPR; SCHW was supported by DFG (SCHW 559/14–1); and DLR was supported by the UNC Bowles Center for Alcohol Studies.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Claudio Da Cunha.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

de Oliveira Guaita, G., Vecchia, D.D., Andreatini, R. et al. Diazepam blocks 50 kHz ultrasonic vocalizations and stereotypies but not the increase in locomotor activity induced in rats by amphetamine. Psychopharmacology 235, 1887–1896 (2018). https://doi.org/10.1007/s00213-018-4878-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00213-018-4878-8

Keywords

  • Diazepam
  • Amphetamine
  • Dopamine
  • Addiction
  • Drugs of abuse
  • Psychostimulants