l-DOPA-induced behavioral sensitization of motor activity in the MPTP-treated common marmoset as a Parkinson's disease model

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Highlights

  • MPTP-treated common marmosets were used as a Parkinson's disease model.

  • Motor activity counts increased gradually by repeated administration of l-DOPA.

  • Behavioral sensitization of above measure accompanied with dyskinesia

  • Behavioral sensitization can be an objective and predictive measure of dyskinesia.

Abstract

l-DOPA is the gold standard for treatment of Parkinson's disease (PD). However, the drug produces some serious side effects, including dyskinesia, which is characterized by repetitive involuntary movements—including chorea. In the present preclinical study using a nonhuman primate model, dyskinesia caused by repeated l-DOPA administration was investigated in the context of behavioral sensitization by objectively quantifying motor activity in the common marmoset of PD model (the Parkinsonian marmoset). Twelve male Parkinsonian marmosets previously treated with MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and six intact male marmosets were used. The motor activity of each marmoset was measured using infrared sensors attached to each individual living cage. Parkinsonian marmosets (n = 6) exhibited behavioral sensitization (enhanced motor activity) in 10 weeks upon oral administration of l-DOPA (10 mg/kg per day on 3 days/week). These animals also exhibited dyskinesia characterized by repetitive rapid movements including chorea in 6–10 weeks. Neither behavioral sensitization nor dyskinesia was observed in Parkinsonian marmosets given vehicle and in intact marmosets given l-DOPA at the same dose (both n = 6 each). Behavioral sensitization was detected sensitively and objectively on motor activity only in Parkinsonian marmosets given repeated l-DOPA at a similar dose used in PD patients. The behavioral feature of the marmosets was dyskinesia similar to that of PD patients but appeared earlier than would be manifested in humans. In spite of statistically significant behavioral sensitization, some marmosets did not exhibit dyskinesia in the present limited l-DOPA administration period. Although both commonalities and differences may exist between behavioral sensitization and dyskinesia, behavioral sensitization is considered to be an objective, quantitative, sensitive and predictive measure of behavioral mechanism underlying dyskinesia in preclinical studies in evaluating compounds.

Introduction

Parkinson's disease (PD) is a movement disorder caused by degeneration of the nigrostriatal dopaminergic neurons of the brain, for reasons that are not understood. These neurons, when healthy, play an important role in smoothing both voluntary and involuntary movements. Thus, degeneration of the neurons has behavioral consequences including immobility, tremor, positional dysfunction, and muscle rigidity, which, together, are termed Parkinsonism or PD-signs. l-DOPA, a therapeutic drug, has served as the “gold standard” for PD treatment for half a century (Fehling, 1966, Fox et al., 2011). However, it was reported in the 1960s that long-term medication of this drug caused development of abnormal and excessive involuntary movements termed dyskinesia (Cotzias et al., 1969, Mones et al., 1971, Treciokas et al., 1971, Huot et al., 2013), as well as other side-effects including wearing-off and on–off phenomena (Lesser et al., 1979, Zappia et al., 1999). Dyskinesia is a collection of various complex abnormal movements characterized by chorea, dystonia, and athetosis (Ellrichmann and Russ, 2007, Jackson et al., 2004) and is distinct from Parkinsonism although the etiology of dyskinesia is not wholly understood.

As definitive knowledge of the causative mechanism of dyskinesia is lacking, much effort has been devoted to develop efficacious PD treatments by drugs that do not have side-effects including dyskinesia (Adamiak et al., 2010, Ahlskog and Muenter, 2001, Muller, 2012, Muller, 2013, Kalinderi et al., 2011, Sprenger and Poewe, 2013). Treatments other than pharmacotherapy have also been employed; these include pallidotomy (Alvarez et al., 2009), deep brain stimulation (Fox et al., 2011, Terzic and Abosch, 2012, Miocinovic et al., 2013), and nerve cell transplantation (Dunnett and Rosser, 2011). Recently, dopamine neurons derived from induced pluripotent stem (iPS) cells are expected to transplant into the brains of PD patients. Such therapy carries great promise, because iPS cells can develop into dopamine neurons and these are unlikely to be rejected by the brain tissues if generated from the stem cells of the same individual (Nishimura and Takahashi, 2013).

In cooperating with the above undertakings, sensitive and valid animal models of PD are essential for preclinical testing of drug candidates to explore possible therapeutic effects with minimum side effects. Nonhuman primates treated with peripheral 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) are generally recognized as valid PD models because the primates develop long-lasting Parkinsonism caused by dopaminergic neural degeneration in the brain (Langston et al., 1984, Fox and Brotchie, 2010). Notably, the MPTP-treated common marmoset, a small primate equivalent in body size to an adult rat, is considered to be a useful, valid, and sensitive model for preclinical evaluation of drug efficacy (Jenner, 2009, Eslamboli, 2005).

The particular characteristics and advantages of the marmoset model have been discussed in detail elsewhere (Ando et al., 2008, Ando et al., 2012). Among them, a normal marmoset is behaviorally active during light-on daytime but not at night showing the similar circadian rhythm as humans but not as rodents. After MPTP administration, daily spontaneous motor activity count in individual living cage measured by a motion sensor decreases markedly and continuously over several months or more. The decreased level of motor activity as a stable behavioral baseline is considered to be an objective and quantitative measure of immobility, one of the main PD-signs. By administration of l-DOPA, for example, the decreased level temporarily increases to normal level at doses comparable to clinical ones. Thus, improving the effect of l-DOPA on immobility could be detected objectively and quantitatively by motor activity measure in combination with subjectively evaluated visual inspection based on score items. At higher doses of l-DOPA, however, motor activity increases much over the normal level with the manifestation of hyper-excitability. Based on these facts, a question arises whether l-DOPA at comparable clinical doses when given repeatedly causes hyper-excitability leading to dyskinesia. If this excitability can be detected as behavioral sensitization of motor activity quantitatively and objectively (Wise and Leeb, 1993), it may help for understanding, measuring and predicting dyskinesia in the preclinical evaluation of new compounds.

The purpose of the present study was to examine behavioral sensitization of motor activity by repeated administration of l-DOPA at comparable clinical dose in the MPTP-treated marmoset and also to examine the relationship between behavioral sensitization and dyskinesia.

Section snippets

Ethics statement

The protocol of the present study was reviewed by the Institutional Animal Care and Use Committee and approved by CIEA (CIEA approval no. 07028A). The criteria used by the committee complied with those mandated by the Japanese Law for the Humane Treatment and Management of Animals. The present study was conducted under the principle of the three Rs (Replacement, Refinement and Reduction) of humane animal experimentation (Balls et al., 1995) and also conducted in strict accordance with the

Experiment 1: repeated l-DOPA administration

In Parkinsonian marmosets, the motor activity count in the first week of oral l-DOPA administration was ~ 6.7-fold (2721.9/408.2) higher than the pre-administration level (Fig. 1) in accordance with alleviated immobility observed by visual inspection. The mean motor activity counts increased gradually upon repeated l-DOPA administration over 10 weeks. The highest mean count was ~ 1.4-fold (3821.0/2721.9) that of week 1, observed in week 8. In Parkinsonian marmosets given vehicle, the mean activity

Summary and its implication

Gradual enhancement of motor activity was observed upon repeated oral administration of l-DOPA at a comparable clinical dose (10 mg/kg: 600 mg per person of 60 kg) to MPTP-treated Parkinsonian marmosets in a shorter term than the clinical one. The enhanced motor activity was not observed in intact marmosets given l-DOPA and in Parkinsonian marmosets given vehicle. Therefore, l-DOPA-induced behavioral sensitization of motor activity was observed only in MPTP-treated Parkinsonian marmosets. In a

Conclusion

The present study quantitated behavioral sensitization induced by repeated l-DOPA in Parkinsonian marmosets. The results form bases for investigation of the mechanism of dyskinesia and indicate that the marmoset model may be sensitive, valid, predictive and useful in preclinical evaluation of drugs and biological products that might cause dyskinesia as one of the serious side effects in PD patients.

The following is the supplementary data related to this article.

Acknowledgments

The authors would like to thank to Ms Sayaka Ohba for her excellent technical assistance and to Dr. Naoko Kagiyama at CIEA for her helpful comments on this manuscript. This study was partly supported by a grant to KA at CIEA from the Neuropsychiatric Disease Project (17-Ko) of the Ministry of Health, Labor and Welfare, Japan. The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding was

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