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Effects of Repeated Treatment with Midazolam in SHR and SLA16 Rat Strains in the Triple Test

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Abstract

We exposed male and female rats of SHR (Spontaneously Hypertensive Rats) and SLA16 (SHR.LEW-Anxrr16) strains, in a non-drugged state, for five consecutive days to the Triple Test (experiment 1); or after repeated treatment with midazolam (MDZ), for four consecutive days. The fifth day was performed without treatment (experiment 2). The first experiment showed that males did not avoid and females increased the exploration of the open arms over the days. In experiment 2, SLA16 from both sexes approached more the open arms than SHR rats. The MDZ anxiolytic-like effect was sustained in both strains and sexes over the days. On the fifth day, SLA16 still approached more the open arms than SHR rats. Data suggest an absence of repeated-trial tolerance to MDZ anxiolytic-like effects. Testing the SHR and SLA16 strains, especially females, could be necessary for the future search for the genes and molecular pathways underlying anxiety/emotionality.

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References

  • Adamec R, Head D, Blundell J, Burton P, Berton O (2006) Lasting anxiogenic effects of feline predator stress in mice: sex differences in vulnerability to stress and predicting severity of anxiogenic response from the stress experience. Physiol Behav 2:12–29

    Article  Google Scholar 

  • Albrechet-Souza L, Borelli KG, Brandao ML (2008) Activity of the medial prefrontal cortex and amygdala underlies one-trial tolerance of rats in the elevated plus-maze. J Neurosci Methods 169:109–118

    Article  Google Scholar 

  • Andreatini R, Vital M, Santos GL (2003) The reinstatement of conflict reverses the one-trial tolerance to diazepam on elevated plus maze. Eur Neuropsychopharmacol 13(Suppl. 4):S378–S379

    Article  Google Scholar 

  • Anselmi M, Correa FJ, Santos JR, Silva AF, Cunha JA, Leão AH, Campêlo CL, Ribeiro AM, Silva RH, Izídio GS (2016) Genetic evidence for chromosome 4 loci influencing learning and memory. Neurobiol Learn Mem 131:182–191

    Article  Google Scholar 

  • Becker JB, Prendergast BJ, Liang JW (2016) Female rats are not more variable than male rats: a meta-analysis of neuroscience studies. Biol Sex Differ 26:7–34

    Google Scholar 

  • Beery AK, Zucker I (2011) Sex bias in neuroscience and biomedical research. Neurosci Biobehav Rev 35:565–572

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2002a) Prior maze experience required to alter midazolam effects in rats submitted to the elevated plus maze. Pharmacol Biochem Behav 72:449–455

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2002b) Anxiolytic effects of ethanol and phenobarbital are abolished in test-experienced rats submitted to the elevated plus maze. Pharmacol Biochem Behav 73(4):963–969

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2003) Anxiolytic-like effects of NMDA/glycine-B receptor ligands are abolished during the elevated plus-maze Trial 2 in rats. Psychopharmacology 170:335–342

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2004) Scopolamine given pre-Trial 1 prevents the one-trial tolerance phenomenon in the elevated plus-maze Trial 2. Behav Pharmacol 15(1):45–54

    Article  Google Scholar 

  • Bertoglio LJ, Joca SR, Guimaraes FS (2006) Further evidence that anxiety and memory are regionally dissociated within the hippocampus. Behav Brain Res 175:183–188

    Article  Google Scholar 

  • Carobrez AP, Bertoglio LJ (2005) Ethological and temporal analyses of anxiety-like behavior: the elevated plus-maze model 20 years on. Neurosci Biobehav Rev 29:1193–1205

    Article  Google Scholar 

  • Carvalho MC, Moreira CM, Zanoveli JM, Brandão ML (2012) Central, but not basolateral, amygdala involvement in the anxiolytic-like effects of midazolam in rats in the elevated plus maze. J Psychopharmacol 26(4):543–554

    Article  Google Scholar 

  • Clayton JA, Collins FS (2014) Policy: NIH to balance sex in cell and animal studies. Nature 509:282–283

    Article  Google Scholar 

  • Cruz-Morales SE, Santos NR, Brandão ML (2002) One trial tolerance to midazolam is due to enhancement of fear and reduction of anxiolytic-sensitive behaviors in the elevated plus-maze retest in the rat. Pharmacol Biochem Behavior 72:973–978

    Article  Google Scholar 

  • Dal-Cól ML, Pereira LO, Rosa VP, Calixto AV, Carobrez AP, Faria MS (2003) Lack of midazolam-induced anxiolysis in the plus-maze Trial 2 is dependent on the length of Trial 1. Pharmacol Biochem Behav 74(2):395–400

    Article  Google Scholar 

  • Davids E, Zhang K, Tarazi FI, Baldessarini RJ (2003) Animal models of attention-deficit hyperactivity disorder. Brain Res Brain Res Rev 2003 42(1):1–21

    Article  Google Scholar 

  • de Araújo Godinho MA, Meneghini L, Lucinda AM, Borges FR, Sakae DY, Marino Neto J, Paschoalini MA, De Lima TC, Faria MS (2007) An approach to evaluate the ability of rats to discriminate different levels of illumination in the plus maze test: effects of scopolamine. Behav Brain Res 4 180(1):86–94

    Article  Google Scholar 

  • De Medeiros GF, Pereira E, Granzotto N, Ramos A (2013) Low-anxiety rat phenotypes can be further reduced through genetic intervention. PLoS ONE 8(12):e83666

    Article  Google Scholar 

  • De Medeiros GF, Corrêa FJ, Corvino ME, Izídio GS, Ramos A (2014) The long way from complex phenotypes to genes: the story of rat chromosome 4 and its behavioral effects. World J Neurosci 4:203–215

    Article  Google Scholar 

  • Díaz-Véliz G, Butrón S, Benavides MS, Dussaubat N, Mora S (2000) Gender, estrous cycle, ovariectomy, and ovarian hormones influence the effects of diazepam on avoidance conditioning in rats. Pharmacol Biochem Behav 66(4):887–892

    Article  Google Scholar 

  • File SE (1990) One-trial tolerance to the anxiolytic effects of chlordiazepoxide in the plus-maze. Psychopharmacology 100:281–282

    Article  Google Scholar 

  • File SE, Zangrossi H (1993) “One-trial tolerance” to the anxiolytic actions of benzodiazepines in the elevated plus-maze, or the development of a phobic state? Psychopharmacology 110:240–244

    Article  Google Scholar 

  • File SE, Zangrossi H Jr, Viana M, Graeff FG (1993) Trial 2 in the elevated plus-maze: a different form of fear? Psychopharmacology 111(4):491–494

    Article  Google Scholar 

  • File SE, Gonzalez LE, Gallant R (1998) Role of the basolateral nucleus of the amygdala in the formation of a phobia. Neuropsychopharmacology 19:397–405

    Article  Google Scholar 

  • File SE, Gonzalez LE, Gallant R (1999) Role of the dorsomedial hypothalamus in mediating the response to benzodiazepines on trial 2 in the elevated plus-maze test of anxiety. Neuropsychopharmacology 21:312–320

    Article  Google Scholar 

  • Fraser LM, Brown RE, Hussin A, Fontana M, Whittaker A, O’Leary TP, Lederle L, Holmes A, Ramos A (2010) Measuring anxiety- and locomotion related behaviours in mice: a new way of using old models. Psychopharmacology 211:99–112

    Article  Google Scholar 

  • Frussa-Filho R, Barbosa-Junior H, Silva RH, Da Cunha C, Mello CF (1999) Naltrexone potentiates the anxiolytic effects of chlordiazepoxide in rats exposed to novel environments. Psychopharmacology (Berl) 147:168–173

    Article  Google Scholar 

  • Gonzalez LE, File SE (1997) A five minute experience in the elevated plus-maze alters the state of the benzodiazepine receptor in the dorsal raphe nucleus. J Neurosci 17:1505–1511

    Article  Google Scholar 

  • Handley SL, Mcblane JW (1993) An assessment of the elevated X-maze for studying anxiety and anxiety-modulating drugs. J Pharmacol Toxicol Methods 29:129–138

    Article  Google Scholar 

  • Hussin AT, Fraser LM, Ramos A, Brown RE (2012) The effect of chlordiazepoxide on measures of activity and anxiety in Swiss–Webster mice in the triple test. Neuropharmacology 63:883–889

    Article  Google Scholar 

  • Izídio GS, Spricigo LJR, Ramos A (2005) Genetic differences in the elevated plus-maze persist after first exposure of inbred rats to the test apparatus. Behav Process 68:129–134

    Article  Google Scholar 

  • Izídio GS, Oliveira LC, Oliveira LFG, Pereira E, Wehrmeister TD, Ramos A (2011) The influence of sex and estrous cycle on QTL for emotionality and ethanol consumption. Mamm Genome 22(5–6):329–340

    Article  Google Scholar 

  • Johnston AL, File SE (1991) Sex differences in animal tests of anxiety. Physiol Behav 49(2):245–250

    Article  Google Scholar 

  • Lister RG (1987) The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology 92:180–185

    PubMed  Google Scholar 

  • Lovick TA (2008) GABA in the female brain-oestrous cycle-related changes in GABAergic function in the periaqueductal grey matter. Pharmacol Biochem Behav 90(1):43–50

    Article  Google Scholar 

  • Maguire J, Mody I (2009) Steroid hormone fluctuations and GABAAR plasticity. Psychoneuroendocrinology 34(1):S84–S90

    Article  Google Scholar 

  • Mccullough LD, de Vries GJ, Miller VM, Becker JB, Sandberg K, Mccarthy MM (2014) NIH initiative to balance sex of animals in preclinical studies. Biol Sex Differ 5:15

    Article  Google Scholar 

  • Pellow S, Chopin P, File SE, Briley M (1985) Validation of open-closed arm entries in an elevated plus-maze as a measure of anxiety in the rat. J Neurosci Methods 14:149–167

    Article  Google Scholar 

  • Pereira JK, Vieira RJ, Konishi CT, Ribeiro RA, Frussa-Filho R (1999) The phenomenon of “one-trial tolerance” to the anxiolytic effect of chlordiazepoxide in the elevated plus-maze is abolished by the introduction of a motivational conflict situation. Life Sci 65(10):101–107

    Article  Google Scholar 

  • Pértile R, Corvino ME, Marchette RCN, Pavesi E, Cavalli J, Ramos A, Izídio GS (2017) The quinpirole hypolocomotive effects are strain and route of administration dependent in SHR and SLA16 isogenic rats. Behav Genet 47(5):552–563

    Article  Google Scholar 

  • Prut L, Belzung C (2003) The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review. Eur J Pharmacol 463:3–33

    Article  Google Scholar 

  • Ramos A, Berton O, Mormede P, Chaouloff F (1997) A multiple-test study of anxiety-related behaviours in six inbred rat strains. Behav Brain Res (85): 57–69

    Article  Google Scholar 

  • Ramos A, Moisan MP, Chaouloff F, Mormède C, Mormède P (1999) Identification of female-specific QTLs affecting an emotionality-related behavior in rats. Mol Psychiatry (4):453–462

    Article  Google Scholar 

  • Ramos A, Pereira E, Martins GC, Wehrmeister TD, Izidio GS (2008) Integrating the open field, elevated plus maze and light/dark box to assess different types of emotional behaviors in one single trial. Behav Brain Res 193(2):277–288

    Article  Google Scholar 

  • Rodgers RJ, Shepherd JK (1993) Influence of prior maze experience on behaviour and response to diazepam in the elevated plus-maze and light/dark tests of anxiety in mice. Psychopharmacology 113(2):237–242

    Article  Google Scholar 

  • Rodgers RJ, Lee C, Sheperd JK (1992) Effects of diazepam on behavioural an antinociceptive responses to the elevated plus-maze in male mice depend upon treatment regimen and prior maze experience. Psychopharmacology 106:102–110

    Article  Google Scholar 

  • Roy V, Chapillon P, Jeljeli M, Caston J, Belzung C (2009) Free versus forced exposure to an elevated plus-maze: evidence for new behavioral interpretations during test and retest. Psychopharmacology 203(1):131–141

    Article  Google Scholar 

  • Sagvolden T (2000) Behavioral validation of the spontaneously hypertensive rat (SHR) as an animal model of attention-deficit/hyperactivity disorder (AD/HD). Neurosci Biobehav Rev 24:31–39

    Article  Google Scholar 

  • Sagvolden T, Russell VA, Aase H, Johansen EB, Farshbaf M (2005) Rodent models of attention-deficit/hyperactivity disorder. Biol Psychiatry 57:1239–1247

    Article  Google Scholar 

  • Schneider P, Ho YJ, Spanagel R, Pawlak CR (2011) A novel elevated plus-maze procedure to avoid the one-trial tolerance problem. Front Behav Neurosci 5:43

    Article  Google Scholar 

  • Stern CAJ, Carobrez AP, Bertoglio LJ (2008) Aversive learning as a mechanism for lack of repeated anxiolytic- like effect in the elevated plus-maze. Pharmacol Biochem Behav 90:545–550

    Article  Google Scholar 

  • Treit D, Menard J, Royan C (1993) Anxiogenic stimuli in the elevated plus-maze. Pharmacol Biochem Behav 44:463–469

    Article  Google Scholar 

  • Tropp J, Markus EJ (2001) Sex differences in the dynamics of cue utilization and exploratory behavior. Behav Brain Res 119(2):143–154

    Article  Google Scholar 

  • Vendruscolo LF, Terenina-Rigaldie E, Raba F, Ramos A, Takahashi RN, Morméde P (2006) Evidence for a female-specific effect of a chromosome 4 locus on anxiety-related behaviors and ethanol drinking in rats. Genes Brain Behav 5(6):441–450

    Article  Google Scholar 

  • Voikar V, Koks S, Vasar E, Rauvala H (2001) Strain and gender differences in the behavior of mouse lines commonly used in transgenic studies. Physiol Behav 72(1–2):271–281

    Article  Google Scholar 

  • Wehrmeister TD, Izídio GS, Pereira E, Izídio G, Ramos A (2010) Absence of repeated-trial tolerance to the anxiolytic-like effects of chlordiazepoxide in the rat triple test. Pharmacol Biochem Behav 97(2):301–309

    Article  Google Scholar 

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Acknowledgements

F. J. Correa and R. C. N. Marchette were recipients of fellowships from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). L. R. Acuña; N. GranzottoF. G. Mazur and E. Corvino were recipients of fellowships from Coordenadoria de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). The authors also thank R. A. N. Pértile and T. M. Alves for their technical assistance.

Funding

This work was supported by Edital MCTI/CNPq 14/2013.

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Authors and Affiliations

  1. Laboratório de Genética do Comportamento, Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, BEG, bloco C, 3° andar, sala 310C, Florianópolis, SC, CEP 88.040-900, Brazil

    Lucía Raily Acuña, Renata Cristina Nunes Marchette, Natalli Granzotto, Paula Gomes Dias, Maria Elisa Corvino, Fernando Gabriel Mazur, Fernanda Junques Corrêa & Geison Souza Izídio

  2. Department of Pharmacology, Federal University of Santa Catarina, Florianópolis, Brazil

    Renata Cristina Nunes Marchette, Natalli Granzotto & Fernanda Junques Corrêa

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  1. Lucía Raily Acuña
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  2. Renata Cristina Nunes Marchette
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  3. Natalli Granzotto
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  5. Maria Elisa Corvino
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  8. Geison Souza Izídio
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Correspondence to Geison Souza Izídio.

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Lucía Raily Acuña, Renata Cristina Nunes Marchette, Natalli Granzotto, Paula Gomes Dias, Maria Elisa Corvino, Fernando Gabriel Mazur, Fernanda Junques Corrêa and Geison Souza Izídio declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were by following the ethical standards of the institution or practice at which the studies were conducted and respected the guidelines of the local committee for Animal Care in Research (CEUA/UFSC) with the valid permission PP00903.

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Acuña, L.R., Marchette, R.C.N., Granzotto, N. et al. Effects of Repeated Treatment with Midazolam in SHR and SLA16 Rat Strains in the Triple Test. Behav Genet 48, 440–450 (2018). https://doi.org/10.1007/s10519-018-9922-2

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