Physiological gait test
an effective method for analyzing balance, locomotion, and neuromuscular disorders in rats and a comparison to the elevated beam test
Keywords:Locomotor activity, Motor coordination, Motor balance, Neurodegenerative diseases, Ivermectin
The evaluation of animal locomotor activity is a behavioral tool widely used to measure the mechanisms underlying a particular disease, disorder, or injury, as well as the effects of exposure to a xenobiotic. The elevated beam test is one of the most used tests in rodents to assess balance and motor coordination. Despite being inexpensive and utilizing a simple apparatus, the high beam test requires a long period of animal training and habituation. The development and characterization of an alternative test, namely the gait test, has the potential to circumvent the time and effort required for animal training, deeming it an effective, inexpensive, and fast method for the analysis of behaviors that are comparably assessed by the high beam test. Therefore, the present study focused on determining the effectiveness and feasibility of the gait test for assessing rodent locomotion and balance as a replacement for the elevated beam test. For this purpose, male rats were divided into three groups: one control group exposed to a saline solution (NaCl 0.9%) and two experimental groups exposed to a single dose of either 0.2 or 1.0 mg/kg of ivermectin intraperitoneally for induction of locomotor disturbance. The high beam and gait tests were performed 15 min and 24 h after drug administration. Results show that the experimental groups had difficulty performing the tasks of either test at both time points analyzed compared to the control groups. At the high beam, experimental animals had trouble maintaining balance and walking. At the gait test, experimental animals showed alterations in gait, which were quantitated by: (a) shortening of step length, (b) decrease of stride, (c) altered step symmetry, and (d) altered stride area. Such results are indicative of compensatory efforts and were comparable between both tests. Altogether, the data indicate that the gait test meets all requirements for assessing motor coordination in rodents. The gait test is therefore validated as a complement to the elevated beam test for the study and analysis of neurodegenerative impairment and other disorders involving neuromuscular disturbances.
Allbutt HN, Henderson JM. Use of the narrow beam test in the rat, 6-hydroxydopamine model of Parkinson’s disease. J Neurosci Methods. 2007;159(2):195-202. http://dx.doi.org/10.1016/j.jneumeth.2006.07.006. PMid:16942799.
Bear MF, Connors BW, Paradiso MA. Neurociências: desvendando o sistema nervoso. 3ª ed. Porto Alegre: Artmed; 2008. 895 p.
Bernardi MM, Kirsten TB, Spinosa HS, Manzano H. Ivermectin impairs sexual behavior in sexually naïve, but not sexually experienced male rats. Res Vet Sci. 2011;91(1):77-81. http://dx.doi.org/10.1016/j.rvsc.2010.07.026. PMid:20800249.
Chiu SHL, Green ML, Baylis FP, Eline D, Rosegay A, Meriwether H, Jacob TA. Absorption, tissue distribution and excretion of tritium-labeled ivermectin in cattle, sheep and rat. J Agric Food Chem. 1990;38(11):2072-8. http://dx.doi.org/10.1021/jf00101a015.
Dunnett SB, Brooks SP. Motor assessment in Huntington’s Disease mice. Methods Mol Biol. 2018;1780:121-41. http://dx.doi.org/10.1007/978-1-4939-7825-0_7. PMid:29856017.
Elble RJ, Cousins R, Leffler K, Hughes L. Gait initiation by patients with lower-half parkinsonism. Brain. 1996;119(Pt 5):1705-16. http://dx.doi.org/10.1093/brain/119.5.1705. PMid:8931591.
Fernagut PO, Diguet E, Labattu B, Tison F. A simple method to measure stride length as an index of nigrostriatal dysfunction in mice. J Neurosci Methods. 2002;113(2):123-30. http://dx.doi.org/10.1016/S0165-0270(01)00485-X. PMid:11772434.
Fukushima AR, Zaccarelli-Magalhães J, Munhoz C, Abreu GR, Camargo ERA, Waziry P, Spinosa HS. Review on requirements for methodological validations and forensic applications. BJFS. 2018;7(4):265-82. http://dx.doi.org/10.17063/bjfs7(4)y2018265.
Hall C. Emotional behavior in the rat: defecation and urination as measures of individual differences in the emotionality. J Comp Physiol. 1934;18(3):385-403.
Hanakawa T, Katsumi Y, Fukuyuma H, Honda M, Hayashi T, Kimura J, Shibasaki H. Mechanisms underlying gait disturbance in Parkinson’s disease: a single photon emission computed tomography study. Brain. 1999;122(7):1271-82. http://dx.doi.org/10.1093/brain/122.7.1271. PMid:10388793.
Hausdorff JM, Cudkowicz ME, Firtion R, Wei JY, Goldberger AL. Gait variability and basal ganglia disorders: stride to stride variations of gait cycle timing in Parkinson’s disease and Huntington’s disease. Mov Disord. 1998;13(3):428-37. http://dx.doi.org/10.1002/mds.870130310. PMid:9613733.
Hikosaka O, Takikawa Y, Kawagoe R. Role of the basal ganglia in the control of purposive saccadic eye movements. Physiol Rev. 2000;80(3):953-78. http://dx.doi.org/10.1152/physrev.2000.80.3.953. PMid:10893428.
Jacobs BY, Kloefkorn HE, Allen KD. Gait analysis methods for rodent models of osteoarthritis. Curr Pain Headache Rep. 2014;18(10):456. http://dx.doi.org/10.1007/s11916-014-0456-x. PMid:25160712.
Machado A. Neuroanotomia funcional. 2ª ed. São Paulo: Atheneu; 2007. 357 p.
Moreira N, Sandini TM, Reis-Silva TM, Navas-Suáresz P, Auada AVV, Lebrun I, Flório JC, Bernardi MM, Spinosa HS. Ivermectin reduces motor coordination, serum testosterone, and central neurotransmitter levels but does not affect sexual motivation in male rats. Reprod Toxicol. 2017;74:195-203. http://dx.doi.org/10.1016/j.reprotox.2017.10.002. PMid:29055807.
Pantaleon L, Fukushima AR, De-Paula LR, Ribeiro GM, Pacca BP, Munoz JWP, Spinosa HS, Ricci EL. Integrative review: verification of the influence of atrazine exposure on behavioral, neurochemical, and Parkinson’s disease disorders. JBRES. 2021;2(10):1044-58. http://dx.doi.org/10.37871/jbres1347.
Pantaleon LP. Influência da exposição à atrazina durante a puberdade na esfera sexual de ratos adultos [dissertation]. São Paulo: University of São Paulo; 2021. 119 p.
Pinto WBC, Souza PVS, Ko GM. Avaliação da marcha normal e patológica no camundongo. RESBCAL. 2015;3(1):45-51.
Rodrigues-Alves PSB. Interferência da moxidectin ana motivação sexual e na ereção peniana de ratos: envolvimento de neurotransmissores hipotalâmicos e estriais adultos [dissertation]. São Paulo: University of São Paulo; 2007. 72 p.
Rodrigues-Alves PSB, Lebrun I, Flório JC, Bernardi MM, Spinosa HS. Moxidectin interference on motor activity of Rats. Braz Arch Biol Technol. 2009;52(4):883-91. http://dx.doi.org/10.1590/S1516-89132009000400012.
Schultz W. Behavioral dopamine signals. Trends Neurosci. 2007;30(5):203-10. http://dx.doi.org/10.1016/j.tins.2007.03.007. PMid:17400301.
Udo MSB. Avaliação dos efeitos tóxicos da exposição pré-natal ao fipronil na prole de ratas Wistar [dissertation]. São Paulo: University of São Paulo; 2021. 143 p.
Wertman V, Gromova A, La Spada AR, Cortes CJ. Low-cost gait analysis for behavioral phenotyping of mouse models of neuromuscular disease. J Vis Exp. 2019;(149). http://dx.doi.org/10.3791/59878-v. PMid:31380846.
Wu T, Hallett M. The cerebellum in Parkinson’s disease. Brain. 2013;136(Pt 3):696-709. http://dx.doi.org/10.1093/brain/aws360. PMid:23404337.
Yamamoto PK, Souza TA, Antiorio ATFB, Zanatto DA, Garcia-Gomes MSA, Alexandre-Ribeiro SR, Oliveira NS, Menck CFM, Bernardi MM, Massironi SMG, Mori CMC. Genetic and behavioral characterization of Kmt2d mouse mutant, a new model for Kabuki Syndrome. Genes Brain Behav. 2019;18(8):e12568. http://dx.doi.org/10.1111/gbb.12568. PMid:30891914.
Zaccarelli-Magalhães J, Sandini TM, Fukushima AR, Spinosa HS. Proposal of open field test as a model of varenicline pharmacokinetic study in rats. Revinter. 2018;11(3):162-9. http://dx.doi.org/10.22280/revintervol11ed3.398.
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