Korean J Physiol Pharmacol.  2013 Feb;17(1):89-97. 10.4196/kjpp.2013.17.1.89.

Differential Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on Motor Behavior and Dopamine Levels at Brain Regions in Three Different Mouse Strains

• Lee KS
• Lee JK
• Kim HG
• Kim HR

• KMID: 1432768
• DOI: http://doi.org/10.4196/kjpp.2013.17.1.89

Abstract

Developing an animal model for a specific disease is very important in the understanding of the underlying mechanism of the disease and allows testing of newly developed new drugs before human application. However, which of the plethora of experimental animal species to use in model development can be perplexing. Administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a very well known method to induce the symptoms of Parkinson's disease in mice. But, there is very limited information about the different sensitivities to MPTP among mouse strains. Here, we tested three different mouse strains (C57BL/6, Balb-C, and ICR) as a Parkinsonian model by repeated MPTP injections. In addition to behavioral analysis, endogenous levels of dopamine and tetrahydrobiopterin in mice brain regions, such as striatum, substantia nigra, and hippocampus were directly quantified by liquid chromatography-tandem mass spectrometry. Repeated administrations of MPTP significantly affected the moving distances and rearing frequencies in all three mouse strains. The endogenous dopamine concentrations and expression levels of tyrosine hydroxylase were significantly decreased after the repeated injections, but tetrahydrobiopterin did not change in analyzed brain regions. However, susceptibilities of the mice to MPTP were differed based on the degree of behavioral change, dopamine concentration in brain regions, and expression levels of tyrosine hydroxylase, with C57BL/6 and Balb-C mice being more sensitive to the dopaminergic neuronal toxicity of MPTP than ICR mice.

Keyword

Dopamine; Mass spectrometry; Mouse strains; MPTP; Parkinson's disease

MeSH Terms

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
Animals
Biopterin
Brain
Dopamine
Dopaminergic Neurons
Hippocampus
Humans
Mass Spectrometry
Mice
Mice, Inbred ICR
Models, Animal
Parkinson Disease
Substantia Nigra
Tyrosine 3-Monooxygenase
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
Biopterin
Dopamine
Tyrosine 3-Monooxygenase

Figure

• 

  Fig. 1 Effects of MPTP (30 mg/kg, s.c) on Rota-rod performance. Animals were trained to stay on an operating Rota-rod for 600 seconds. On the test day, animals were evaluated on the Rota-rod 6 and 24 hours after MPTP administration. (A) MPTP (C57BL/6, 30 mg/kg, s.c.), (B) MPTP (ICR, 30 mg/kg, s.c.), (C) MPTP (C57BL/6, ICR, Balb C, 20 mg/kg, s.c.), (D) body weight after MPTP administration. Data are expressed as mean latency to fall off the Rota-rod (vertical bars represent standard error of the mean). *p<0.05, **p<0.01 vs 0 hour (naïve) group, #p<0.05 compared between 6 hours and 24 hours groups by Student's t-test, n=6~10.

• 

  Fig. 2 Summary of experimental design. Each groups received MPTP treatment for 3 days followed by one MPTP-free day. At the end of behavioral test of day 4, animals were euthanized for brain sample collection.

• 

  Fig. 3 Effect of MPTP (20 mg/kg, s.c) on moving distance for 1 hour following MPTP administration. (A) C57BL/6 mice, (B) Balb C mice, (C) ICR mice. Data are expressed as accumulated total moving distances in cages (vertical bars represent standard error of the mean). *p<0.05, **p<0.01, ##p<0.01 vs day 0 (n=6~10).0

• 

  Fig. 4 Effect of MPTP (20 mg/kg, s.c) on total rearing frequencies until 1 hour after MPTP administration. (A) C57BL/6 mice, (B) Balb C mice, (C) ICR mice. Data are expressed as accumulated number of rearing frequencies in cages (vertical bars represent standard error of the mean). *p<0.05, **p<0.01 vs day 0 (n=6~10).

• 

  Fig. 5 Comparison of moving distances and total rearing frequencies among three mouse strains Effect of MPTP (20 mg/kg, s.c) on moving distances (A) and total rearing frequencies (B) until 1 hour after MPTP administration over three days. In each case, the mean results were normalized by the mean of day 0. Vertical bars represent standard error of the mean and negative SE bars have been deleted for clarity. Two-way ANOVA, *p<0.05, **p<0.01 vs ICR (n=6~10).

• 

  Fig. 6 Effects of MPTP (20 mg/kg, s.c) on dopamine concentration in mouse brain section. The endogenous level of dopamine in mice brain scetions ((A) striatum, (B) substantia nigra, (C) hippocampus) were measured by LC-MS/MS in three different mice species. (D) internal calibration for dopamine. Data is expressed as mean±s.e.m. *p<0.05, **p<0.01 (n=5).

• 

  Fig. 7 Effects of MPTP (20 mg/kg, s.c) on tetrahydrobiopterin (BH4) concentration in brain regions. The endogenous level of BH4 in mice brain regions ((A) striatum, (B) substantia nigra, (C) hippocampus) were measured by LC-MS/MS in three different mice strains. (D) Internal calibration for BH4. Data is expressed as mean±standard error of the mean (n=5).

• 

  Fig. 8 Effects of MPTP (20 mg/kg, s.c) on the expression of tyrosine hydroxylase in the striatum in the three strains of mice. (A) Representative immunoblots of tyrosine hydroxylase (TH) and β-actin expression. (B) Densitometry plot of TH expression immunoblots normalized to the control levels, with and without MPTP treatment (n=4). *p<0.05 and **p<0.01 vs. black bar (control group).

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