Vitamin C supplementation promotes locomotor and exploratory behaviors in male Wistar rats exposed to varying stress models


Vitamin C,
Explorative Behavior
Locomotor Behavior,
Oxidative stress

How to Cite

Olumide, O., Aderemi, T., Arikawe , A. P., Idowu, A. J., Olatunji-Bello, I., & Umoren, A. (2023). Vitamin C supplementation promotes locomotor and exploratory behaviors in male Wistar rats exposed to varying stress models. Nigerian Journal of Physiological Sciences, 38(1), 57–64.


Constant exposure to environmental stress has negative behavioral outcomes. Considering the inverse relationship between stress and Vitamin C intake, this study was aimed at investigating variable stress techniques and Vitamin C supplementation on exploratory/locomotor behaviors in male Wistar rats. Twenty-eight male Sprague-Dawley rats (100g-120g) were allotted into four groups (n=7). Control received 10ml/kg distilled water, group two received 100 mg/kg vitamin C, group three was exposed to different models of stress while group four was stressed alongside 100 mg/kg vitamin C. Vitamin C treatments were given orally for 2 weeks. Animals in groups 3 and 4 were stressed every other day with models such as multiple cage changes, exposure to noise, overnight strange objects, overnight wetting of beddings, and immobility. Explorative and locomotor activities were assessed with the open field test, novel object recognition test, and Y maze test using a Logitech camera and ANY-maze software to track the movement of the rats. Cortisol was assayed in the serum using Enzyme-linked Immuno Assay (ELISA) kit. Superoxide Dismutase, catalase, and lipid peroxidase; malondialdehyde (MDA) were also assayed in the serum. The results show that locomotor activities such as distance traveled, average speed, and time spent in the center square was significantly reduced by stress. These activities were improved with the intake of vitamin C compared with stress. Explorative activities such as locomoting around the environment, orientating towards novelty, and touching or sniffing novel objects were significantly increased in the rats on Vitamin C supplements and reduced in the stressed group. In the serum, cortisol level was significantly increased in rats exposed to stress and decreased with Vitamin C intake. Stress also significantly increased MDA and decreased SOD and CAT while vitamin C supplement decreased MDA and increased SOD and CAT. In conclusion, oral intake of vitamin C enhanced explorative/locomotor behavior and increased oxidative stress in rats exposed to different models of stress


Aboul-Ela, H.M., Saad, A.A., El-Sikaily, A.M.A. et al. (2011). Oxidative stress and DNA damage in relation to transition metals overload in Abu-Qir Bay, Egypt, J. Genetic Engineering and Biotechnol. 9(1):51-58.

Adeneye, A.A and Olagunjun, J.O (2009). Protective Effect of Oral Ascorbic Acid (Vitamin C) on Acetaminophen-Induced Renal Injury in Rats. African Journal of Biomedical Research, Volume 12(1): 51-61.

Aebi, H. Catalase in vitro. (1984). Methods Enzymol. 105:121–6.

Aguilar, T.A.F., Navarro, B.C.H., & Pérez, J.A.M. (2016). Endogenous Antioxidants: A Review of their Role in Oxidative Stress. A Master Regulator of Oxidative Stress - The Transcription Factor Nrf2. doi: 10.5772/65715.

Angelow, S., Haselbach, M., Galla, H.J. (2003). Functional characterisation of the active ascorbic acid transport into cerebrospinal fluid using primary cultured choroid plexus cells. Brain Res. 988:105–113.

Blossom, V., Gokul, M., Kumar, N.A. et al. (2020) Chronic unpredictable stress-induced inflammation and quantitative analysis of neurons of distinct brain regions in Wistar rat model of comorbid depression, Veterinary World, 13(9): 1870-1874.

Borrow, A.P., Bales, N.J., Stover, S.A. et al. (2018). Chronic Variable Stress Induces Sex-Specific Alterations in Social Behavior and Neuropeptide Expression in the Mouse, Endocrinology, 159(7):2803–2814,

Borta, A., & Schwarting, R.K.W. (2005). Inhibitory avoidance, pain reactivity, and plus-maze behavior in Wistar rats with high versus low rearing activity. Physiology & Behavior, 84(3), 387–396. doi:10.1016/j.physbeh.2005.01.009.

Buege, J.A. and Aust, S.D. (1978). Microsomal lipid peroxidation. Methods in Enzymol. 52:302-310.

Crosswell, A.D and Lockwood, K.G. (2020). Best practices for stress measurement: How to measure psychological stress in health research. Health Psychology Open.7 (2).

Edwards, J.A., Webster, S., VanLaar, D. et al. (2008). Psychometric analysis of the UK Health and Safety Executive’s Management Standards work-related stress Indicator Tool. Work Stress. 2008: 22:96-107.

Gaweł, S., Wardas, M., Niedworok, E. et al. (2004). Malondialdehyde (MDA) as a lipid peroxidation marker. J. Wiad Lek. 57(9-10):453-5.

Harrison, F.E., & May, J.M. (2009). Vitamin C function in the brain: vital role of the ascorbate transporter SVCT2. Free Radic Biol Med. 15;46(6):719-30. doi: 10.1016/j.freeradbiomed.2008.12.018. Epub 2009 Jan 6. PMID: 19162177; PMCID: PMC2649700.

Idowu, A.J., Osima, E.O., Umoren, G.A. (2019). Motor and Cognitive Outcomes of Environmental Enrichment during Development in Male BALB/c mice. LASU Journal of Medical Sciences, 4(2); 14-19.

Jaggi, A.S., Bhatia, N., Kumar, N. et al. (2011). A review on animal models for screening potential anti-stress agents. Neurol Sci 32: 993–1005.

Kalueff, A.V., Stewart, A.M., Song, C. et al. (2016). Neurobiology of rodent self-grooming and its value for translational neuroscience Nat Rev Neurosci. 17(1):45-59. doi:10.1038/nrn.2015.8.

Khassaf, M., McArdle, A., Esanu, C. et al. (2003). Effect of vitamin C supplements on antioxidant defence and stress proteins in human lymphocytes and skeletal muscle. The Journal of physiology, 549(Pt 2), 645–652.

Kozler, P., Maresova, D., Pokorny, J. (2017). Study of locomotion, rearing and grooming activity after single and/or concomitant lesions of central and peripheral nervous system in rats. Neuro Endocrinol Lett. 38 (7):495-501.

Lightman, S.L., Birnie, M.T., Conway-Campbell, B.L. (2020). Dynamics of ACTH and Cortisol Secretion and Implications for Disease. Endocr Rev.1;41(3):bnaa002. doi: 10.1210/endrev/bnaa002. PMID: 32060528; PMCID: PMC7240781.

Mällo, T., Alttoa, A., Kõiv, K. et al. (2007). Rats with persistently low or high exploratory activity: behaviour in tests of anxiety and depression, and extracellular levels of dopamine. Behav. Brain Res. 27;177 (2):269-81. doi: 10.1016/j.bbr.2006.11.022.

Marik, P.E. (2020). Vitamin C: an essential "stress hormone" during sepsis. J Thorac Dis. 12(1):S84-S88.

Matisz, C.E., Badenhorst, C.A., Gruber, A.J. (2021). Chronic unpredictable stress shifts rat behavior from exploration to exploitation. Stress. 24(5):635-644. doi: 10.1080/10253890.2021.1947235. Epub 2021 Jul 5. PMID: 34223804.

Meerlo, P., Sgoifo, A., Turek, F.W. (2002). The effects of social defeat and other stressors on the expression of circadian rhythms. Stress. 5:15–22.

Mirończuk-Chodakowska, I., Witkowska, A.M., Zujko, M.E. (2018). Endogenous non-enzymatic antioxidants in the human body. Adv. Med. Sci. 63(1):68-78. doi: 10.1016/j.advms.2017.05.005.

Moritz, B., Schmitz, A.E., Rodrigues, A.L.S. et al. (2020). The role of vitamin C in stress-related disorders. J Nutr. Biochem. 85:108459. doi: 10.1016/j.jnutbio.2020.108459. Epub 2020 Jul 3. PMID: 32745879.

Moussa, Z., M.A. Judeh, Z., & A. Ahmed, S. (2020). Nonenzymatic Exogenous and Endogenous Antioxidants. Free Radical Medicine and Biology. doi: 10.5772/intechopen.87778.

Mu, M.D., Geng, H.Y., Rong, K.L. et al. (2020). A limbic circuitry involved in emotional stress-induced grooming. Nat Commun 11; 2261.

Mustafi S., & Wang G. (2020). Vitamin C: epigenetic role and cancer. Molecular Nutri. 691-709.

Nelson, R.J. (2005). An introduction to behavioral endocrinology. 3rd ed. Sinauer Associates; Sunderland, MA: 2005b. Biological rhythms; pp. 586–667.

Oueghlani, Z, Simonnet, C., Cardoit, L. et al. (2018). Brainstem Steering of Locomotor Activity in the Newborn Rat. J Neurosci. 29; 38(35):7725-7740. doi: 10.1523/JNEUROSCI.1074-18.2018.

Padayatty, S. J., & Levine, M. (2016). Vitamin C: the known and the unknown and Goldilocks. Oral diseases, 22(6), 463–493.

Podhorna, J. & Brown, R.E. (2002). Strain differences in activity and emotionality do not account for differences in learning and memory performance between C57BL/6 and DBA/2 mice. Genes Brain Behav. 1(2):96-110. doi: 10.1034/j.1601-183x.2002.10205.x.

Rojas-Carvajal, M., Chinchilla-Alvarado, J., Brenes, J.C. (2022). Muscarinic regulation of self-grooming behavior and ultrasonic vocalizations in the context of open-field habituation in rats. Behav Brain Res. 10; 418:113641. doi: 10.1016/j.bbr.2021.113641.

Salami, S.A., Salahdeen, H.M., Moronkola, O.T. et al. (2020). Vitamin C supplementation during chronic variable stress exposure modulates contractile functions of testicular artery and sperm parameters in male Wistar rats. Middle East Fertil. Soc. J 25: 8.

Sgroi, S., Kaelin-Lang, A., Capper-Loup, C. (2014). Spontaneous locomotor activity and L-DOPA-induced dyskinesia are not linked in 6-OHDA parkinsonian rats. Front in Behav Neurosci. 2;8:331. doi: 10.3389/fnbeh.2014.00331.

Sharma, S., Advani, D., Das, A. et al. (2022). Pharmacological intervention in oxidative stress as a therapeutic target in neurological disorders, Journal of Pharmacy and Pharmacology 74(4); 461–484,

Sik, A., van Nieuwehuyzen, P., Prickaerts, J. et al. (2003). Performance of different mouse strains in an object recognition task. Behav Brain Res. 147(1-2):49-54. doi: 10.1016/s0166-4328(03)00117-7.

Smolinsky, A.N., Bergner, C.L., LaPorte, J.L. et al. (2009). Analysis of Grooming Behavior and Its Utility in Studying Animal Stress, Anxiety, and Depression. Neuromethods 42:21-36.

Srivastava, K. K., & Kumar, R. (2015). Stress, oxidative injury and disease. Indian journal of clinical biochemistry: IJCB, 30(1), 3–10.

Sun, M. and Zigman, S. (1978). An improved spectrophotometric assay for superoxide dismutase based on epinephrine autoxidation. J Anal. Biochem. 90:81–9.

Tucker, J., Sinclair, R., Mohr C. et al. (2008). A temporal investigation of the direct, interactive, and reverse relations between demand and control and affective strain, Work & Stress. 22: 81-95.

Zimcikovaa E., Simkob J, Karesovac I. et al. (2017). Behavioral effects of antiepileptic drugs in rats: Are the effects on mood and behavior detectable in open-field test? Seizure 52:35–40.

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Copyright (c) 2023 Nigerian Journal of Physiological Sciences