JoVE Logo
Faculty Resource Center

Sign In

Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Behavior

Behavioral Assessments of Spontaneous Locomotion in a Murine MPTP-induced Parkinson's Disease Model

Published: January 7th, 2019

DOI:

10.3791/58653

1State Key Laboratory of Medicinal Chemical Biology, The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Science, Nankai University

We describe establishment of a murine model for Parkinson's disease using MPTP, and behavioral assessments using cylinder and open field tests to measure motor function. We then use L-DOPA as one example to show how to apply this model in the study of PD drugs.

Parkinson’s disease (PD) is a common neurodegenerative disorder disease, causing the phenomenon of shaking, rigidity, slowness of movement and dementia. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) can lead to some Parkinson’s-like symptoms by destroying dopaminergic neurons in the substantia nigra of the brain. It has been thus used to establish PD models in various animal studies. Here, mice receive MPTP injections (20 mg/kg/day) for seven days and the behavioral tests are performed on the eighth day. This model is adapted efficiently in the study of PD. The behavioral tests here include the cylinder test and the open field test. The cylinder experiment is used to detect the animals’ ability to lift their front paws when put into a different environment. As the PD model mice show arching—the mouse arches its back—the number of paw liftings decrease. This test is easy to execute. The open field test is used to detect the amount of time the mice spend on running, walking, and remaining immobile. We analyze animals’ movements in open field using software and obtain data. Lastly, we use L-DOPA, one of the most commonly used PD drugs, as one example to show how to apply this model to the study of PD drugs. Our results indicate that MPTP neurotoxicity induces motor deficit which can be mitigated by L-DOPA.

Parkinson's disease (PD), one of the most common diseases among older individuals, is a long-term neurodegenerative disorder1. Patients always show the phenomenon of shaking, rigidity, slowness of movement and dementia that worsen over time2. Other symptoms including sensory, sleep, and emotional problems are also commonly observed2. The cause of PD is still unclear, but it is generally believed to involve both genetic and environmental factors, which induce loss of dopaminergic neurons in the substantia nigra3, and development of Lewy bodies and Lewy neurite....

Log in or to access full content. Learn more about your institution’s access to JoVE content here

This study was performed according to the international, national and institutional rules considering animal experiments. The study protocol was approved by the animal ethics committee of Nankai University.

1. MPTP and L-DOPA administration

NOTE: Ten-week-old female BALB/c mice were provided by the Institute of Zoology, Chinese Academy of Sciences. Mice were housed six per cage under a 12 h light/dark cycle (lights on at 08:00-20:00), a constant temperature of 21-22 &.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

In the cylinder test, the decrease of rears against the wall was observed in mice (group 2) treated with MPTP from day 1 to day 7 and saline on day 8 as compared with saline-treated mice (group 1), while an increase of rears was observed in the mice (group 3) treated with MPTP from day 1 to day 7 and L-DOPA on day 8 as compared with the mice (group 2) treated with MPTP from day 1 to day 7 and saline on day 8 (Figure 1).

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Due to destruction of dopaminergic neurons in the substantia nigra of the brain, MPTP causes Parkinson's-like symptoms in mice7. L-DOPA is the most preferred drug for PD ever since its clinical use, because it helps in maintaining normal daily activities in patients with PD, with effective suppression of motor abnormalities including akinesia and rigidity15. The mice treated with MPTP showed impairments in the behavioral tests like cylinder test and open field test, whi.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Our work is funded by Tianjin Undergraduate Training Programs for Innovation and Entrepreneurship (Grant No. 63183004). This project was initiated in the State Key Laboratory of Medicinal Chemical Biology at Nankai University. The authors declare that there are no conflicts of interests.

....

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Name Company Catalog Number Comments
70% Ethanol Ruiboxing Company RBX-64175
Camera BASLER acA645-100gm
Cylinder test Made in-house at Nankai University N/A
Excel Microsoft N/A
Levodopa Sigma-Aldrich 72816
Matalb 2017a Mathworks N/A
Mice Institute of Zoology, Chinese Academy of Sciences Balb/c Adult female mice(10 weeks)
MPTP Yuanye Biological Technology Company Ltd., Shanghai S31504-500mg
Open field test Made in-house at Nankai University N/A
Syringe Solelybio S-xsgwz-w Irrigation
Syringe Jiangxi Fenglin Medical Application Co. hc3824 Intraperitoneal injection

  1. Dehay, B., Bourdenx, M., Gorry, P., Przedborski, S., Vila, M., Hunot, S., Singleton, A., Olanow, C. W., Merchant, K. M., Bezard, E. Targeting -synuclein for treatment of Parkinson's disease: mechanistic and therapeutic considerations. Lancet Neurology. 14, 855-866 (2015).
  2. Weintraub, D., Comella, C., Horn, L., S, Parkinson's disease - Part 1: Pathophysiology, symptoms, burden, diagnosis, and assessment. American Journal of Managed Care. 14, 40-48 (2008).
  3. Ismaiel, A. A., et al. Metformin, besides exhibiting strong in vivo anti-inflammatory properties, increases mptp-induced damage to the nigrostriatal dopaminergic system. Toxicology and Applied Pharmacology. , 19-30 (2016).
  4. Bayliss, J. A., et al. Ghrelin-AMPK signaling mediates the neuroprotective effects of calorie restriction in Parkinson's disease. Journal of Neuroscience. 36, 3049-3063 (2016).
  5. Olanow, C. W., Stern, M. B., Sethi, K. The scientific and clinical basis for the treatment of Parkinson disease. Neurology. 72 (21 Supplement 4), (2009).
  6. Chiba, K., Trevor, A., Castagnoli, N. Metabolism of the neurotoxic tertiary amine, MPTP, by brain monoamine oxidase. Biochemical & Biophysical Research Communications. 120 (2), 574-578 (1984).
  7. Bakay, R. A., et al. Biochemical and behavioral correction of MPTP parkinson-like syndrome by fetal cell transplantation. Annals of the New York Academy of Sciences. 495 (1), 623-638 (2010).
  8. Kelly, P. H., Seviour, P. W., Iversen, S. D. Amphetamine and apomorphine responses in the rat following 6-OHDA lesions of the nucleus accumbenssepti and corpus striatum. Brain Research. 94 (3), 507-522 (1975).
  9. Langston, J. W., Irwin, I., Langston, E. B., Forno, L. S. 1-methyl-4-phenylpyridinium ion (MPP+): identification of a metabolite of MPTP, a toxin selective to the substantia nigra. Neuroscience Letters. 48 (1), 87-92 (1984).
  10. Katila, N., et al. Metformin lowers α-synuclein phosphorylation and upregulates neurotrophic factor in the MPTP mouse model of Parkinson's disease. Neuropharmacology. 125, (2017).
  11. Goldberg, N. R. S., Haack, A. K., Lim, N. S., Janson, O. K., Meshul, C. K. Dopaminergic and behavioral correlates of progressive lesioning of the nigrostriatal pathway with MPTP. Neuroscience. 180, 256-271 (2011).
  12. Schwarting, R. K., Sedelis, M., Hofele, K., Auburger, G. W., Huston, J. P. Strain-dependent recovery of open-field behavior and striatal dopamine deficiency in the mouse MPTP model of Parkinson's disease. Neurotoxicity Research. 1 (1), 41 (1999).
  13. Shiguang, S., et al. The open field test as a method for ethology in Kunming mice: test-retest reliability. Chinese Journal of Behavioral Medicine and Brain Science. 19 (12), 1093-1095 (2010).
  14. Roome, R. B., Vanderluit, J. L. Paw-Dragging: a Novel, Sensitive Analysis of the Mouse Cylinder Test. Journal of Visualized Experiments. (98), e52701 (2015).
  15. Cotzias, G. C., Van Woert, M. H., Schiffer, L. M. Aromatic amino acids and modification of Parkinsonism. New England Journal of Medicine. 282, 31-33 (1967).
  16. Cassady, J. C., Johnson, R. E. Cognitive test anxiety and academic performance. Contemporary Educational Psychology. 27 (2), 270-295 (2002).
  17. Rogalewski, A., et al. Semaphorin 6A improves functional recovery in conjunction with motor training after cerebral ischemia. Plos One. 5 (5), 0010737 (2010).
  18. Chen, W., Qiao, D., Liu, X., Shi, K. Treadmill exercise improves motor dysfunction and hyperactivity of the corticostriatal glutamatergic pathway in rats with 6-ohda-induced parkinson's disease. Neural Plasticity. 2017, 2583910 (2018).
  19. Colas-Zelin, D., et al. The imposition of, but not the propensity for, social subordination impairs exploratory behaviors and general cognitive abilities. Behavioural Brain Research. 232 (1), 294-305 (2012).
  20. Spurney, C., et al. Preclinical drug trials in the mdx mouse: Assessment of reliable and sensitive outcome measures. Muscle Nerve. 39, 591-602 (2009).
  21. Valle, F. P. Effects of strain, sex, and illumination on open-field behavior of rats. American Journal of Psychology. 83, 103-111 (1970).
  22. Tatem, K. S., et al. Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases. Journal of Visualized Experiments. (91), e51785 (2014).

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved