Sex differences in the elevated plus-maze test and large open field test in adult Wistar rats.
Open accessAbstract
There is a growing need for a better understanding of sex differences in animal models of psychiatric disorders. The elevated plus-maze (EPM) test and large open field (LOF) test are widely used to study anxiety-like behavior in rodents. Our studies explored sex differences in anxiety and activity parameters in the LOF and EPM and determined whether these parameters correlate within and between tests. Drug naïve adult male and female Wistar rats (n = 47/sex) were used for the studies, and the rats were tested for 5 min in the EPM and 10 min in the LOF. The females spent more time on the open arms of the EPM and made more open arms entries than the males. The females also spent more time in the center zone of the LOF and made more center zone entries. The females traveled a greater distance in the LOF and EPM. There was a moderate positive correlation between time on the open arms of the EPM and time in the center zone of the LOF. There was also a moderate positive correlation between open arms entries in the EPM and center zone entries in the LOF. A hierarchical cluster analysis revealed one cluster with LOF parameters, one cluster with EPM parameters, and one cluster with parameters related to the avoidance of open spaces. In conclusion, these findings indicate that female rats display less anxiety-like behavior in the EPM and LOF. Furthermore, there are sex differences for almost all behavioral parameters in these anxiety tests.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The sex effects in the OFT are in agreement with recent findings on sex differences in the OFT.
“The sex effects observed in the OFT were indicative of lower anxiety in females, and this is in agreement with recent findings on sex differences in the OFT ( Knight et al., 2021).”
From Stage 2 Registered Report: Refinement of tickling protocols to improve positive animal welfare in laboratory rats (2025) · cites this paper as Knight et al., 2021
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
ggplot2: Elegant Graphics for Data Analysis
Wickham, H. (2009) · doi:10.1007/978-0-387-98141-3 · Cited by 14,447 · see citing works
McLean, C. P., Asnaani, A., Litz, B. T., & Hofmann, S. G. (2011) · Journal of Psychiatric Research · doi:10.1016/j.jpsychires.2011.03.006 · Cited by 2,519 · see citing works
Gene–environment interactions: early life stress and risk for depressive and anxiety disorders
Nugent, N. R., Tyrka, A. R., Carpenter, L. L., & Price, L. H. (2011) · Psychopharmacology · doi:10.1007/s00213-010-2151-x · Cited by 269 · see citing works
Meerlo, P., Overkamp, G. J. F., Benning, M. A., Koolhaas, J. M., & Van Den Hoofdakker, R. H. (1996) · Physiology & Behavior · doi:10.1016/0031-9384(95)02271-6 · Cited by 151 · see citing works
Pohl, J., Olmstead, M. C., Wynne-Edwards, K. E., Harkness, K., & Menard, J. L. (2007) · Behavioral Neuroscience · doi:10.1037/0735-7044.121.3.462 · Cited by 120 · see citing works
Sex differences in social interaction in rats: role of the immediate-early gene Zif268
Stack, A., Carrier, N., Dietz, D., Hollis, F., Sorenson, J., & Kabbaj, M. (2010) · Neuropsychopharmacology · doi:10.1038/npp.2009.163 · Cited by 113 · see citing works
Flagged first, then by citations
Research works printed but not linked: 58
- Albrechet-Souza L, Schratz CL, Gilpin NW, 2020. Sex differences in traumatic stress reactivity in rats with and without a history of alcohol drinking. Biol Sex Differ 11(1), 27– 27. 32393336 PMC7216391
- Asami T, Yamasue H, Hayano F, Nakamura M, Uehara K, Otsuka T, Roppongi T, Nihashi N, Inoue T, Hirayasu Y, 2009. Sexually dimorphic gray matter volume reduction in patients with panic disorder. Psychiatry Research: Neuroimaging 173(2), 128– 134. 19560907
- Beery AK, Zucker I, 2011. Sex bias in neuroscience and biomedical research. Neuroscience & Biobehavioral Reviews 35(3), 565– 572. PMC3008499 20620164
- Blizard DA, Lippman HR, Chen JJ, 1975. Sex differences in open-field behavior in the rat: the inductive and activational role of gonadal hormones. Physiology & behavior 14(5), 601– 608. 1135316
- Bruijnzeel AW, Knight P, Panunzio S, Xue S, Bruner MM, Wall SC, Pompilus M, Febo M, Setlow B, 2019. Effects in rats of adolescent exposure to cannabis smoke or THC on emotional behavior and cognitive function in adulthood. Psychopharmacology 236, 2773– 2784. 31044291 PMC6752736
- Bruijnzeel AW, Qi X, Guzhva LV, Wall S, Deng JV, Gold MS, Febo M, Setlow B, 2016. Behavioral Characterization of the Effects of Cannabis Smoke and Anandamide in Rats. PloS one 11(4), e0153327. 27065006 PMC4827836
- Burke NN, Coppinger J, Deaver DR, Roche M, Finn DP, Kelly J, 2016. Sex differences and similarities in depressive-and anxiety-like behaviour in the Wistar-Kyoto rat. Physiology & behavior 167, 28– 34. 27591842
- Cahill L, 2006. Why sex matters for neuroscience. Nature reviews neuroscience 7(6), 477. 16688123
- Dagan SY, Tsoory MM, Fainzilber M, Panayotis N, 2016. COLORcation: a new application to phenotype exploratory behavior models of anxiety in mice. Journal of neuroscience methods 270, 9– 16. 27269190
- Dalla C, Pitychoutis PM, Kokras N, Papadopoulou-Daifoti Z, 2010. Sex differences in response to stress and expression of depressive-like behaviours in the rat, Biological basis of sex differences in psychopharmacology. Springer, pp. 97– 118. 21769725
- de Jongh R, Geyer MA, Olivier B, Groenink L, 2005. The effects of sex and neonatal maternal separation on fear-potentiated and light-enhanced startle. Behavioural brain research 161(2), 190– 196. 15878207
- Dichter GS, Brunelli SA, Hofer MA, 1996. Elevated plus-maze behavior in adult offspring of selectively bred rats. Physiology & Behavior 60(1), 299– 304. 8804680
- Domonkos E, Borbélyová V, Csongová M, Bosý M, Kačmárová M, Ostatníková D, Hodosy J, Celec P, 2017. Sex differences and sex hormones in anxiety-like behavior of aging rats. Horm Behav 93, 159– 165. 28576648
- Doremus-Fitzwater TL, Varlinskaya EI, Spear LP, 2009. Effects of pretest manipulation on elevated plus-maze behavior in adolescent and adult male and female Sprague-Dawley rats. Pharmacology, biochemistry, and behavior 92(3), 413– 423. PMC3064078 19344672
- Elliott BM, Faraday MM, Phillips JM, Grunberg NE, 2004. Effects of nicotine on elevated plus maze and locomotor activity in male and female adolescent and adult rats. Pharmacol.Biochem.Behav 77(1), 21– 28. 14724038
- Fraley C, Raftery AE, 1998. How many clusters? Which clustering method? Answers via model-based cluster analysis. The computer journal 41(8), 578– 588.
- Galili T, 2015. dendextend: an R package for visualizing, adjusting and comparing trees of hierarchical clustering. Bioinformatics 31(22), 3718– 3720. 26209431 PMC4817050
- Genn RF, Tucci SA, Thomas A, Edwards JE, File SE, 2003. Age-associated sex differences in response to food deprivation in two animal tests of anxiety. Neuroscience & Biobehavioral Reviews 27(1–2), 155– 161. 12732231
- Gentsch C, Lichtsteiner M, Feer H, 1987. Open field and elevated plus-maze: a behavioural comparison between spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats and the effects of chlordiazepoxide. Behavioural brain research 25(2), 101– 107. 3675823
- Goodwin RD, Faravelli C, Rosi S, Cosci F, Truglia E, de Graaf R, Wittchen H-U, 2005. The epidemiology of panic disorder and agoraphobia in Europe. European Neuropsychopharmacology 15(4), 435– 443. 15925492
- Gray JA, Lalljee B, 1974. Sex differences in emotional behaviour in the rat: correlation between open-field defecation and active avoidance. Animal Behaviour 22, 856– 861. 4477944
- Gué M, Bravard A, Meunier J, Veyrier R, Gaillet S, Recasens M, Maurice T, 2004. Sex differences in learning deficits induced by prenatal stress in juvenile rats. Behavioural brain research 150(1–2), 149– 157. 15033288
- Harrington GM, 1972. Strain differences in open-field behavior of the rat. Psychonomic Science 27(1), 51– 53.
- Hogg S, 1996. A review of the validity and variability of the elevated plus-maze as an animal model of anxiety. Pharmacology, biochemistry, and behavior 54(1), 21– 30. 8728535
- Imhof JT, Coelho ZM, Schmitt ML, Morato GS, Carobrez AP, 1993. Influence of gender and age on performance of rats in the elevated plus maze apparatus. Behavioural brain research 56(2), 177– 180. 8240712
- Johnston AL, File SE, 1991. Sex differences in animal tests of anxiety. Physiology & behavior 49(2), 245– 250. 2062894
- Johnston AL, File SE, 1991. Sex differences in animal tests of anxiety. Physiol Behav 49(2), 245– 250. 2062894
- Kessler RC, McGonagle KA, Zhao S, Nelson CB, Hughes M, Eshleman S, Wittchen H-U, Kendler KS, 1994. Lifetime and 12-month prevalence of DSM-III-R psychiatric disorders in the United States: results from the National Comorbidity Survey. Archives of general psychiatry 51(1), 8– 19. 8279933
- Korte SM, De Boer SF, 2003. A robust animal model of state anxiety: fear-potentiated behaviour in the elevated plus-maze. European journal of pharmacology 463(1–3), 163– 175. 12600708
- Lenroot RK, Giedd JN, 2010. Sex differences in the adolescent brain. Brain and cognition 72(1), 46– 55. 19913969 PMC2818549
- Liebsch G, Montkowski A, Holsboer F, Landgraf R, 1998. Behavioural profiles of two Wistar rat lines selectively bred for high or low anxiety-related behaviour. Behavioural brain research 94(2), 301– 310. 9722280
- Liu S, Seidlitz J, Blumenthal JD, Clasen LS, Raznahan A, 2020. Integrative structural, functional, and transcriptomic analyses of sex-biased brain organization in humans. Proceedings of the National Academy of Sciences 117(31), 18788– 18798. PMC7414084 32690678
- Maeng LY, Milad MR, 2015. Sex differences in anxiety disorders: interactions between fear, stress, and gonadal hormones. Hormones and behavior 76, 106– 117. 25888456 PMC4823998
- Mansouri MT, Fidler JA, Meng QC, Eckenhoff RG, Garcia PS, 2019. Sex effects on behavioral markers of emergence from propofol and isoflurane anesthesia in rats. Behavioural brain research 367, 59– 67. 30898682
- Mirzasoleiman B, Badanidiyuru A, Karbasi A, Vondrák J, Krause A, 2015. Lazier than lazy greedy, Twenty-Ninth AAAI Conference on Artificial Intelligence.
- Pavlova IV, Broshevitskaya ND, Onufriev MV, Moiseeva YV, 2020. Sex-Related Differences in Anxious-Depressive and Defensive Behavior in Wistar Rats. Neuroscience and Behavioral Physiology 50(9), 1163– 1175.
- 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(3), 149– 167. 2864480
- Pijlman FT, Wolterink G, Van Ree JM, 2003. Physical and emotional stress have differential effects on preference for saccharine and open field behaviour in rats. Behavioural brain research 139(1–2), 131– 138. 12642184
- Prut L, Belzung C, 2003. The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review. European journal of pharmacology 463(1–3), 3– 33. 12600700
- Qi X, Guzhva L, Yang Z, Febo M, Shan Z, Wang KK, Bruijnzeel AW, 2016. Overexpression of CRF in the BNST diminishes dysphoria but not anxiety-like behavior in nicotine withdrawing rats. Eur Neuropsychopharmacol 26, 1378– 1389. 27461514 PMC5067082
- Ramos A, Kangerski AL, Basso PF, Da Silva Santos JE, Assreuy J, Vendruscolo LF, Takahashi RN, 2002. Evaluation of Lewis and SHR rat strains as a genetic model for the study of anxiety and pain. Behavioural brain research 129(1–2), 113– 123. 11809502
- Ramos A, Pereira E, Martins GC, Wehrmeister TD, Izídio GS, 2008. Integrating the open field, elevated plus maze and light/dark box to assess different types of emotional behaviors in one single trial. Behavioural brain research 193(2), 277– 288. 18590774
- Renard GM, Suárez MM, Levin GM, Rivarola MA, 2005. Sex differences in rats: effects of chronic stress on sympathetic system and anxiety. Physiol Behav 85(3), 363– 369. 15939444
- Rylkova D, Shah HP, Small E, Bruijnzeel AW, 2009. Deficit in brain reward function and acute and protracted anxiety-like behavior after discontinuation of a chronic alcohol liquid diet in rats. Psychopharmacology (Berl) 203(3), 629– 640. 19048233 PMC3138076
- Sakhaie N, Sadegzadeh F, Mohammadnia A, Dadkhah M, Saadati H, 2020. Sex‐dependent effects of post‐weaning exposure to an enriched environment on novel objective recognition memory and anxiety‐like behaviors: the role of hippocampal BDNF level. International Journal of Developmental Neuroscience. 32416621
- Scholl JL, Afzal A, Fox LC, Watt MJ, Forster GL, 2019. Sex differences in anxiety-like behaviors in rats. Physiology & behavior 211, 112670. 31487491
- Shansky RM, 2019. Are hormones a “female problem” for animal research? Science 364(6443), 825– 826. 31147505
- Soares-Cunha C, Coimbra B, Borges S, Domingues AV, Silva D, Sousa N, Rodrigues AJ, 2018. Mild Prenatal Stress Causes Emotional and Brain Structural Modifications in Rats of Both Sexes. Frontiers in behavioral neuroscience 12, 129. 30034328 PMC6043801
- Steenbergen HL, Farabollini F, Heinsbroek RP, 1991. Sex-dependent effects of aversive stimulation on holeboard and elevated plus-maze behavior. Behavioural brain research 43(2), 159– 165. 1867757
- Tan S, Xue S, Behnood-Rod A, Chellian R, Wilson R, Knight P, Panunzio S, Lyons H, Febo M, Bruijnzeel AW, 2019. Sex differences in the reward deficit and somatic signs associated with precipitated nicotine withdrawal in rats. Neuropharmacology 160, 107756. 31487496 PMC6842125
- Turano A, Osborne BF, Schwarz JM, 2018. Sexual differentiation and sex differences in neural development. 29967999
- Walker DL, Toufexis DJ, Davis M, 2003. Role of the bed nucleus of the stria terminalis versus the amygdala in fear, stress, and anxiety. Eur.J.Pharmacol 463, 199– 216. 12600711
- Wang T, Han W, Chitre AS, Polesskaya O, Woods LCS, Palmer AA, Chen H, 2018. Social and anxiety-like behaviors contribute to nicotine self-administration in adolescent outbred rats. Scientific reports 8(1), 18069. 30584246 PMC6305389
- Ward JH Jr, 1963. Hierarchical grouping to optimize an objective function. Journal of the American statistical association 58(301), 236– 244.
- Whitley E, Ball J, 2002. Statistics review 2: Samples and populations. Critical Care 6(2), 1– 6. 11983040 PMC137296
- Winther G, Elfving B, Müller HK, Lund S, Wegener G, 2018. Maternal High-fat Diet Programs Offspring Emotional Behavior in Adulthood. Neuroscience 388, 87– 101. 30025860
- Yang R, Sun H, Wu Y, Lu G, Wang Y, Li Q, Zhou J, Sun H, Sun L, 2019. Long-lasting sex-specific effects based on emotion-and cognition-related behavioral assessment of adult rats after post-traumatic stress disorder from different lengths of maternal separation. Frontiers in Psychiatry 10, 289. 31231246 PMC6558979
- Zanni G, DeSalle MJ, Deutsch HM, Barr GA, Eisch AJ, 2020. Female and male rats readily consume and prefer oxycodone to water in a chronic, continuous access, two-bottle oral voluntary paradigm. Neuropharmacology 167, 107978. 32001238 PMC9748519
Cited by
Sorted by citations
Marquardt, A. E., VanRyzin, J. W., Fuquen, R. W., & McCarthy, M. M. (2023) · Frontiers in Behavioral Neuroscience · doi:10.3389/fnbeh.2022.1076765 · Cited by 31 · see citing works
“However, replicating other studies (Johnston and File, 1991; Knight et al., 2021), we observed a small but significant sex difference in the LDB animals with a trending effect in the SDB animals (p = 0.”
ResultPossamai-Della, T., Cararo, J. H., Aguiar-Geraldo, J. M., Peper-Nascimento, J., Zugno, A. I., Fries, G. R., Quevedo, J., & Valvassori, S. S. (2023) · Molecular Neurobiology · doi:10.1007/s12035-023-03348-1 · Cited by 16 · see citing works
Bombail, V., Brown, S. M., Beyts, C., Martin, J. E., Li, T., Meddle, S. L., Mendl, M., Robinson, E. S., Hammond, T. J., Nielsen, B. L., et al. (2025) · F1000Research · doi:10.12688/f1000research.166417.2 · Cited by 4 · see citing works
- Authors
- Knight, P., Chellian, R., Wilson, R., Behnood-Rod, A., Panunzio, S., & Bruijnzeel, A. W.
- Affiliations
- University of Florida (US)
- Corresponding author
- Bruijnzeel, Adriaan W.
- Venue
- Pharmacology Biochemistry and Behavior
- Publisher
- Elsevier BV
- Published
- 2021-05
- Volume
- 204
- Pages
- 173168
- ISSN
- 0091-3057 (ISSN-L)
- PMC
- PMC8130853
- OpenAlex
- W3133727976
- Cited by
- 252
- References
- 6
- Citations by year
- 2026: 51 · 2025: 68 · 2024: 46 · 2023: 50 · 2022: 28 · 2021: 9
- FWCI
- 15.39
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Pharmacology Biochemistry and Behavior (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- University of Florida
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurotransmitter Receptor Influence on Behavior
- Social Sciences › Psychology › Experimental and Cognitive Psychology › Anxiety, Depression, Psychometrics, Treatment, Cognitive Processes
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- sex differences, anxiety-like behavior, elevated plus maze, Wistar rats, locomotor activity, female rats, animal models of psychiatric disorders
- MeSH terms
- Open Field Test, Elevated Plus Maze Test, Animals, Anxiety, Behavior, Animal, Disease Models, Animal, Female, Male, Motor Activity, Sex Characteristics, Rats, Wistar, Maze Learning, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
