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Keywords:

  • dopamine;
  • DYT1 dystonia;
  • HPLC;
  • in vivo microdialysis;
  • mouse;
  • striatum

Abstract

  1. Top of page
  2. Abstract
  3. Methods
  4. Results
  5. Discussion
  6. Acknowledgement
  7. References

Early onset torsion dystonia, the most common form of hereditary primary dystonia, is caused by a mutation in the TOR1A gene, which codes for the protein torsinA. This form of dystonia is referred to as DYT1. We have used a transgenic mouse model of DYT1 dystonia [human mutant-type (hMT)1 mice] to examine the effect of the mutant human torsinA protein on striatal dopaminergic function. Analysis of striatal tissue dopamine (DA) and metabolites using HPLC revealed no difference between hMT1 mice and their non-transgenic littermates. Pre-synaptic DA transporters were studied using in vitro autoradiography with [3H]mazindol, a ligand for the membrane DA transporter, and [3H]dihydrotetrabenazine, a ligand for the vesicular monoamine transporter. No difference in the density of striatal DA transporter or vesicular monoamine transporter binding sites was observed. Post-synaptic receptors were studied using [3H]SCH-23390, a ligand for D1 class receptors, [3H]YM-09151-2 and a ligand for D2 class receptors. There were again no differences in the density of striatal binding sites for these ligands. Using in vivo microdialysis in awake animals, we studied basal as well as amphetamine-stimulated striatal extracellular DA levels. Basal extracellular DA levels were similar, but the response to amphetamine was markedly attenuated in the hMT1 mice compared with their non-transgenic littermates (253 ± 71% vs. 561 ± 132%, < 0.05, two-way anova). These observations suggest that the mutation in the torsinA protein responsible for DYT1 dystonia may interfere with transport or release of DA, but does not alter pre-synaptic transporters or post-synaptic DA receptors. The defect in DA release as observed may contribute to the abnormalities in motor learning as previously documented in this transgenic mouse model, and may contribute to the clinical symptoms of the human disorder.

Abbreviations used
D1 and D2

DA receptors

DA

dopamine

DAT

DA transporter

DOPAC

dihydroxyphenyl acetic acid

hMT

human mutant-type

HVA

homovanillic acid

s. a.

specific activity

VMAT2

vesicular monoamine transporter

Dystonia is a disorder characterized by sustained muscle contractions, frequently causing twisting and repetitive movements, or abnormal postures (Jankovic and Fahn 1993). The most common type of early onset generalized dystonia is an autosomal dominant disorder caused by a 3-bp deletion (ΔGAG) in the TOR1A gene that encodes torsinA (Ozelius et al. 1997). Symptoms of this disorder typically develop before the age of 21 years and include involuntary sustained muscle contractions that cause abnormal posturing of a foot, leg or arm, which frequently generalize to other body regions (Bressman et al. 2002). The pathophysiologic mechanisms, by which the ΔGAG mutation causes dystonia is unclear. There is no evidence for neurodegeneration in the limited number of postmortem examinations of DYT1 patients that have been conducted, although enlargement of dopamine (DA) neurons (Rostasy et al. 2003) and ubiquitin inclusions (McNaught et al. 2004) have been reported. The affected protein, torsinA, shares homology with members of the AAA + ATPase gene family (Lupas et al. 1997; Neuwald et al. 1999). Members of this family participate in numerous cellular functions, including protein folding and degradation, membrane trafficking and vesicle fusion (Ogura and Wilkinson 2001).

Genetic and pharmacological studies have led to the hypothesis that at least some forms of dystonia arise from disturbances in striatal dopaminergic signaling. Mutations within the tyrosine hydroxylase gene (Knappskog et al. 1995) and the GTP cyclohydrolase gene (Ichinose et al. 1994), both of which limit DA synthesis, result in a dystonic phenotype in humans. In addition, mechanical or ischemic lesions of the striatum and administration of pharmacological agents that block DA D2 receptors in vivo can result in a dystonic phenotype (Rupniak et al. 1986,Skidmore and Reich 2005). Several types of hereditary dystonia have Parkinsonian features and early onset Parkinson’s disease can present as dystonia (Leung et al. 2001; Klien et al. 2007). In human brain, the mRNA for torsinA is strongly expressed in dopaminergic neurons of the substantia nigra, although it is also found in other regions of the brain including the hippocampus and cerebellum (Augood et al. 1999). Positron emission tomography has demonstrated reduced binding of D2 ligands in human carriers of the DYT1 mutation (Asanuma et al. 2005). Our laboratory conducted a study, which we directly examined DA metabolism in postmortem human brain from three individuals with the DYT1 mutation and found no change in striatal DA content, but there was an alteration in the ratio of dihydroxyphenyl acetic acid (DOPAC) to DA (Augood et al. 2004). This alteration suggests a change in DA turnover, but the number of cases available for study was small and thus the strength of this conclusion is limited.

A transgenic mouse model which reproduces some features of the human disorder has been described previously (Sharma et al. 2005). These mice express either human wild-type or human mutant-type (hMT) torsinA under the control of a constitutive cytomegalovirus promoter, as well as endogenous wild-type mouse torsinA under its endogenous promoter. They do not have overt dystonia, but do demonstrate a defect in motor learning comparable with that seen in human non-manifesting carriers of the DYT1 mutation (Sharma et al. 2005). We have used one of these mouse models (hMT1 mice) to examine the effect of the hMT torsinA protein on tissue levels of DA and its metabolites in the striatum using HPLC with electrochemical detection. We also examined pre-synaptic DA transporters [both DA transporter (DAT) and vesicular monoamine transporter (VMAT2)] and post-synaptic DA receptors (D1 and D2 class) using in vitro ligand binding methods. Moreover, we examined basal and amphetamine-stimulated striatal extracellular DA levels in these animals using in vivo microdialysis. Our data show that presence of the hMT torsinA does not alter the tissue content of DA or total binding densities of pre-synaptic transport sites or post-synaptic striatal DA receptors, but does result in impaired release of DA after amphetamine treatment. These observations suggest that the mutation of torsinA protein may interfere with stimulated striatal DA signaling, and that this defect may contribute both to the motor learning defect previously documented in this mouse model, as well as to the clinical symptoms of the disorder in humans.

Methods

  1. Top of page
  2. Abstract
  3. Methods
  4. Results
  5. Discussion
  6. Acknowledgement
  7. References

Animals

Human mutant-type 1 transgenic mice expressing mutant-type human torsinA driven by the cytomegalovirus promotor were generated as described previously (Sharma et al. 2005). A total of 51 mice were used and housed with food and water ad libitum under controlled conditions (12 h light/dark cycle). Mice were genotyped prior to the experiments, and a second confirmatory genotyping was conducted at the end of each experiment.

Autoradiography

All studies were performed on coded samples by experimenters blinded to genotype. All assays were also conducted at a single saturating concentration of each ligand and these ligands strongly labeled the striatum. Six-month-old female transgenic DYT1 mice were compared with their age-matched non-transgenic female littermates. Brains were rapidly frozen in liquid isopentane. Sections (12 μm) containing striatum and cortex were mounted onto polylysine coated glass slides, kept at −70°C, and thawed 1 h prior to use. Assays for DAT, VMAT2, D1-like and D2-like DA receptors were performed as described previously (Cha et al. 1998). [3H]ligands for the assays are as follows; [3H]mazindol for DAT, [3H]-dihydrotetrabenazine for VMAT2, [3H]SCH-23390 for D1-like DA receptors, [3H]YM-09151–2 for D2-like DA receptors. Slides were apposed to tritium-sensitive film (Hyperfilm 3H, Amersham, Piscataway, NJ, USA), with calibrated radioactive standards, for 2–3 weeks. Films were developed and analyzed using a computer-based image analysis system (M1, Imaging Research, St. Catharine’s, ON, Canada). Image density corresponding to binding of [3H]ligand was converted to pmol/mg protein by using calibrated radioactive standards, and non-specific binding was subtracted.

For DAT binding, slides were pre-washed for 5 min in cold buffer: 50 mmol/L Tris–HCl, 5 mmol/L KCl, 300 mmol/L NaCl, pH 7.9. Slides were then incubated in 6 nmol/L [3H]mazindol (specific activity (s.a). 24 Ci/mmol) in the presence of 300 nmol/L desipramine for 2.5 h. Non-specific binding was defined in the presence of 10 μmol/L nomifensine. For VMAT2 binding, slides were pre-washed for 20 min in 25°C buffer: 50 mmol/L Na Phosphate, pH 7.7. Slides were then incubated in 5 nmol/L [3H]dihydrotetrabenazine (s.a. 20 Ci/mmol) for 40 min at 25°C. Non-specific binding was defined in the presence of 2 μmol/L tetrabenazine. Assays for D1-like and D2-like DA receptors used a buffer containing 25 mmol/L Tris–HCl (pH 7.5), 100 mmol/L NaCl, 1 mmol/L MgCl2, 1 μmol/L pargyline, and 0.001% ascorbate. For D1-like receptors, slides were incubated with 1.65 nmol/L [3H]SCH-23390 (s.a. 70.3 Ci/mmol) for 2.5 h. Non-specific binding was defined in the presence 1 μmol/L cis-flupentixol. For D2-like receptors, slides were incubated with 180 pmol/L [3H]YM-09151–2 (s.a. 85.5 Ci/mmol) for 3 h. Non-specific binding was defined in the presence 50 μmol/L DA.

Slides were then rinsed twice in cold buffer for 3 min, rinsed quickly in distilled water, and dried under a stream of cool air and were apposed to tritium-sensitive film (Hyperfilm 3H, Amersham) with calibrated radioactive standards and allowed to expose for 2–3 weeks. Films were developed and analyzed using a computer-based image analysis system (M1, Imaging Research). Image density corresponding to binding of [3H]ligand was converted to pmol/mg protein by using calibrated radioactive standards, and non-specific binding was subtracted.

Surgical procedures

Microdialysis experiments followed similar procedures established earlier (Rouge-Pont et al. 2002). Briefly, microdialysis studies were performed on six-month-old male transgenic DYT1 mice and their non-transgenic male littermates. All studies were performed on coded samples by experimenters blinded to genotype. Mice were anesthetized with ketamine/xylazine (100/10 mg/kg) and implanted unilaterally with a microdialysis guide cannula (CMA/7) aimed at the striatum using standard stereotaxic techniques with coordinates in millimeter: anterior 0.6, lateral 1.9 relative to bregma, and ventral 2.0 from the dura surface that were based on the atlas of Franklin and Paxinos (1997). The microdialysis guide cannula and a head-mount screw, which permitted attachment of the animal to a spring tether, were cemented in place with dental acrylic. Animals were allowed to recover for 7–10 days before the microdialysis experiments.

Microdialysis procedures

The night before the experiment, the animal was brought into the experiment chamber and a microdialysis probe (CMA/7; membrane dimension 0.24 × 2 mm) was manually inserted into the guide cannula and perfused with artificial cerebrospinal fluid containing (in mmol/L): 147 NaCl, 2.7 KCl, 1 MgCl2, 1.2 CaCl2, 2.0 Na2HPO4; pH 7.4 ± 2. The inlet tubing of the probe was connected to a microinfusion pump via a dual quartz-lined swivel set at a flow rate of 1 μL/min, and the animal was placed into the Plexiglas test chamber. The animals were allowed to habituate to the microdialysis environment overnight. This habituation period also allowed levels of DA to return to baseline levels after the probe insertion. The following day, four consecutive 20 min samples were collected for the determination of basal levels of DA. Baseline is defined as the average of these four samples. After the collection of baseline samples, animals received saline vehicle (10 cc/kg, i.p.) and one sample was collected every 20 min for the next 100 min. Then they received amphetamine in saline (5 mg/kg, i.p.) and additional samples were collected for the following 100 min. Samples were stored at −80°C until analyzed using HPLC-electrochemistry as described previously (Balcioglu et al. 2003). At the end of each experiment, animals were perfused with formaldehyde (4% in saline) and brains were removed and sectioned into 50 μm thick sections using vibratome. Sections were stained with Nissl for anatomical observations. Only animals with correct probe placement were used in data analysis.

Tissue dissection

Striatal DA content was studied in a separate group of adult male hMT1 mice and littermate controls (= 6 in each group). The right striatum of each animal was dissected, weighed, and placed in a separate tube containing 0.4 N perchloric acid (1 mL/100 μg tissue). The tissues were homogenized separately and then centrifuged at 13 000 g for 20 min at 4°C. Supernatants were separated, filtered, and analyzed for DA along with its metabolites by HPLC-electrochemistry as described previously (Balcioglu et al. 2003).

Statistics

For autoradiography experiments, tissue DA, and extracellular basal DA measurements, an unpaired t-test was used. For microdialysis experiments, two-way anova was used to evaluate the effects of treatment and genotype followed by post hoc tests using the Bonferroni method. Significance level was < 0.05. Prism 4 software was used to conduct these analyses (Graphpad Inc., San Diego, CA, USA).

Results

  1. Top of page
  2. Abstract
  3. Methods
  4. Results
  5. Discussion
  6. Acknowledgement
  7. References

Striatal tissue dopamine levels

Analysis of the DA and its metabolites, homovanillic acid (HVA) and DOPAC, revealed a similar trend towards higher content in the hMT1 mice, but these differences did not reach statistical significance. No difference in the ratios of HVA to DA or DOPAC to DA was observed (Table 1).

Table 1.   Tissue levels of DA and its metabolites (DOPAC and HVA) in the striatum of hMT1 mice and their non-transgenic littermates
GroupDADOPACHVADOPAC/DAHVA/DA
  1. Data are average ± SEM for = 6 for each group. Data are normalized to milligram of tissue weight and unit is nmol/mg tissue weight. Difference between groups for any of the above parameters is not significant at the level of p < 0.05.

hMT1 mice48 ± 975 ± 144 ± 0.41.56 ± 0.20.08 ± 0.04
Non-transgenic litter mates34 ± 653 ± 153 ± 11.55 ± 0.20.09 ± 0.01

Pre-synaptic dopamine uptake sites

Analysis of the films revealed no difference in the density of [3H]mazindol (a ligand for DAT) or [3H]dihydrotetrabenazine (a ligand for VMAT2) sites between the hMT1 mice and their non-transgenic littermates ([3H]mazindol: hMT1, 2.13 ±0.80 pmol/mg protein; non-transgenic littermates, 1.93 ±0.65 pmol/mg protein; [3H]dihydrotetrabenazine: hMT1, 0.78 ± 0.10 pmol/mg protein; non-transgenic littermates, 0.82 ± 0.07 pmol/mg protein. = 5 for each group, differences are not significant).

Striatal dopamine receptor sites

Analysis of the autoradiograms revealed no difference in the density of [3H]SCH-23390 (a ligand for D1 class receptors) or [3H]YM-09151–2 (a ligand for D2 class receptors) binding sites between the hMT1 mice and their non-transgenic littermates ([3H]SCH-23390: hMT1, 2.39 ± 0.42 pmol/mg protein; non-transgenic littermates, 1.99 ± 0.24 pmol/mg protein; [3H]YM-09151–2: hMT1, 0.47 ± 0.03 pmol/mg protein; non-transgenic littermates, 0.46 ± 0.04 pmol/mg protein. = 5 for each group, differences are not significant).

Striatal extracellular dopamine

There was no difference in the mean baseline levels of extracellular DA (corrected for the in-vitro recovery of probes) between the hMT1 mice and their non-transgenic littermates [hMT1, 164 ± 79 nmol/L (= 6); non-transgenic littermates, 178 ± 79 nmol/L (= 9)]. To assess the effect of drug treatments on DA release, data from each animal were normalized to the corresponding pre-treatment baseline extracellular DA level and expressed as a percent of baseline. Neither the hMT1 mice nor their non-transgenic littermates exhibited a significant response to the saline vehicle injection (hMT1, 111 ± 18%; their non-transgenic littermates, 116 ± 9% of baseline). In the non-transgenic littermates, amphetamine produced a marked enhancement of striatal extracellular DA (561 ± 132%, = 9), but this effect was substantially attenuated in the hMT1 mice (253 ± 71%, = 7, p < 0.05 by anova) (Fig. 1).

image

Figure 1.  Amphetamine-stimulated striatal extracellular dopamine (DA) levels in the human mutant-type (hMT)1 mice and their non-transgenic controls. All mice were treated systemically with saline (10 cc/kg, i.p.) and amphetamine (5 mg/kg, i.p). Data are normalized to the pre-treatment basal concentration in each animal. Mean amphetamine-stimulated DA levels were 253 ± 71% in the hMT1 mice and 561 ± 132% in their non-transgenic littermates. Difference between groups for amphetamine-stimulated DA levels is significant at the level of p < 0.05. = 7 for hMT1 mice and = 9 for their non-transgenic littermates.

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Discussion

  1. Top of page
  2. Abstract
  3. Methods
  4. Results
  5. Discussion
  6. Acknowledgement
  7. References

Using a mouse model of human DYT1 dystonia, we observed that expression of human mutant torsinA leads to an impairment of amphetamine-induced DA release in the striatum. This effect cannot be attributed to an impairment in the synthesis or storage of DA, as no reduction in the striatal content of DA or its metabolites was observed; indeed, the data revealed a trend towards an increased content of DA, DOPAC and HVA in the hMT1 mice. In addition, we found no significant differences in the density of pre-synaptic DA uptake sites (DAT or VMAT2), or post-synaptic striatal DA receptors between transgenic mice and their non-transgenic littermates. These observations suggest that the primary defect in DA metabolism produced by the presence of the mutant form of the human torsinA protein is an abnormality of DA release.

As recently reviewed by Sulzer et al. (2005), amphetamine produces DA release by displacing DA from pre-synaptic vesicles. This leads to elevated cytoplasmic DA levels in pre-synaptic terminals, and release into the extracellular space by reversal of the normal action of the DAT. This is of interest because several lines of evidence have pointed to an interaction between torsinA and expression of VMAT and DAT. In HEK293 cells, co-expression of torsinA with DAT reduces [3H]-DA uptake by decreasing the level of DAT present at the plasma membrane (Torres et al. 2004). In Caenorhabditis elegans, over-expression of the torsinA homolog Tor2 leads to down-regulation of DAT expression (Cao et al. 2005), while in SH-SY5Y neuroblastoma cells over-expression of mutant, but not wild-type, torsinA leads to entrapment of VMAT and torsinA in membranous structures (Misbahuddin et al. 2005). In this study, we used binding of [3H]-mazindol to study DAT sites and [3H]-dihydrotetrabenazine to study VMAT2 sites in the striatum of our transgenic mouse model (hMT1 mice). We found that neither [3H]-mazindol sites nor [3H]-dihydrotetrabenazine sites were altered by the expression of human mutant torsinA. However, it is important to note that this assay does not distinguish between surface and intracellular ligand binding sites. Thus, it is possible that the altered release of DA in response to amphetamine in these animals arises from an altered surface expression of DAT or altered subcellular distribution of VMAT2, which were not detected using the ligand binding assays.

In vivo microdialysis is the most direct way to assess extracellular DA in an intact animal, but it is necessarily an invasive technique and there is a possibility that local tissue injury contributes to measurement variability (Westerink 1995; Kuczenski et al. 1997; Carboni et al. 2001). The variability of DA release in response to amphetamine, which we observed is typical of this type of experiment, and the magnitude of inhibition of DA release in the hMT1 mice is similar to that seen with other manipulations of dopaminergic neuron function, such as knockout of GABAB1 receptor function (Vacher et al. 2006).

Several other mouse models of DYT1 dystonia have been described. In a model in which the mutant torsinA protein was over-expressed under control of a neuron specific enolase promoter (Shashidharan et al. 2005), a variable phenotype was observed with some animals exhibiting hyperactivity, while others are behaviorally normal. In these animals, tissue levels of DA were differentially altered, with a small increase in the non-symptomatic animals and decrease in symptomatic mice, while levels of DOPAC and HVA were not altered. In another DYT1 mouse model in which endogenous knock-in ΔGAG mutation was introduced, hyperactivity and impaired beam walking were observed (Dang et al. 2005). In these knock-in animals, tissue levels of DA were not different between groups but there was a slight reduction in HVA. Similar results were obtained in a knock-down mouse model with reduced expression of endogenous torsinA, which did not alter DA content but did slightly reduce DOPAC content (Dang et al. 2006). In the limited studies of human DYT1 dystonia that have been reported, no change in striatal DA content has been observed (Augood et al. 2004;Furukawa et al. 2000). Collectively, these data support the view that in animals as well as humans, neither the presence of mutant torsinA nor a partial deficiency of the normal protein has a substantial effect on the synthesis or storage of DA in striatal pre-synaptic terminals. Our data suggest that the major defect in our animal model is in the dynamic properties of DA release, as demonstrated by the impairment in amphetamine-stimulated release, which we have observed. It will be important to determine whether a similar abnormality of release is present in the other mouse models of DYT1 dystonia which have been described.

The mouse model we have studied does not have overt dystonia, but does demonstrate a reduced ability to learn motor skills in an accelerating rotarod paradigm (Sharma et al. 2005). This defect may be similar to the abnormalities observed in the ‘non-manifesting carrier’ state of the human disorder. Only about 30% of those who carry the DYT1 mutation develop dystonia, but those who carry the gene but lack overt symptoms nevertheless have demonstrable abnormalities, which include a defect in motor learning ability and an altered pattern of cerebral recruitment when learning new motor tasks (Ghilardi et al. 2003). DA release is essential for motor learning, and DA impairment is associated with impaired rotarod performance in rodents (Ogura et al. 2005 and Mizoguchi et al. 2002). Thus, impaired DA release similar to that which we have observed in animals expressing the human mutant torsinA protein may contribute to the motor learning abnormalities of human carriers of the DYT1 mutation.

The identification of dopaminergic abnormalities in animal models of DYT1 dystonia is important, because this defect may contribute to clinical symptoms of the disorder and it suggests alternative therapeutic approaches to this disorder. If in fact presence of the mutant form of torsinA in humans leads to dysregulation of DA release, then strategies to augment release or even therapies directed at activation of post-synaptic DA receptors may prove beneficial.

Acknowledgement

  1. Top of page
  2. Abstract
  3. Methods
  4. Results
  5. Discussion
  6. Acknowledgement
  7. References

This study was supported by NIH grant NS37409.

References

  1. Top of page
  2. Abstract
  3. Methods
  4. Results
  5. Discussion
  6. Acknowledgement
  7. References
  • Asanuma K., Ma Y., Okulski J., Dhawan V., Chaly T., Carbon M., Bressman S. B. and Eidelberg D. (2005) Decreased striatal D2 receptor binding in non-manifesting carriers of the DYT1 dystonia mutation. Neurology 64, 347349.
  • Augood S. J., Martin D. M., Ozelius L. J., Breakefield X. O., Penney J. B. Jr and Standaert D. G. (1999) Distribution of the mRNA encoding torsinA abd torsinB in the normal adult human brain. Ann. Neurol. 46, 761769.
  • Augood S. J., Hollingsworth Z., Albers D. S., Yang L., Leung J., Breakefield X. O. and Standaert D. G. (2004) Dopamine transmission in DYT1 dystonia. Adv. Neurol. 94, 5360.
  • Balcioglu A., Zhang K. and Tarazi F. I. (2003) Dopamine depletion abolishes apomorphine and amphetamine-induced increases in extracellular serotonin levels in the striatum of conscious rats. A microdialysis study. Neuroscience 119, 10451053.
  • Bressman S. B., Raymond D., Wendt K., Saunders-Pullman R., De leon D., Fahn S., Ozelius L. and Risch N. (2002) Diagnostic criteria for dystonia in DYT1 families. Neurology 59, 17801782.
  • Cao S., Gelwix C. C., Caldwell K. A. and Caldwell G. A. (2005) Torsin-mediated protection from cellular stress in the dopaminergic neurons of Caenorhabditis elegans. J. Neurosci. 25, 38013812.
  • Carboni E., Spielewoy C., Vacca C., Nosten-Bertrand M., Giros B. and Di Chiara G. (2001) Cocaine and amphetamine increase extracellular dopamine in the nucleus accumbens of mice lacking the dopamine transporter gene. J. Neurosci. 21, 14.
  • Cha J. J., Kosinski C. M., Kerner J. A., Alsdorf S. A., Mangiarini L., Davies S. W., Penney J. B., Bates G. P. and Young A. B. (1998) Altered brain neurotransmitter receptors in transgenic mice expressing a portion of an abnormal human Huntington disease gene. Proc. Natl Acad. Sci. USA 95, 64806485.
  • Dang M. T., Yokoi F., McNaught K. P., Jengelley T. A., Jackson T., Li J. and Li Y. (2005) Generation and characterization of Dyt1 λGAG knock-in mouse as a model for early-onset dystonia. Exp. Neurol. 196, 452463.
  • Dang M. T., Yokoi F., Pence M. A. and Li Y. (2006) Motor deficits and hyperactivity in Dyt1 knockdown mice. Neurosci. Res. 56, 470474.
  • Franklin K. B. and Paxinos G. (1997) The Mouse Brain in Stereotaxic Coordinates. Academic press, Burlington, MA, USA.
  • Furukawa Y., Hornykiewicz O., Fahn S. and Kish S. J. (2000) Striatal dopamine in early-onset primary torsin dystonia with the DYT1 mutation. Neurology 54, 11931195.
  • Ghilardi M. R., Carbon M., Silvestri G., Dhawan V., Tagliati M., Bressman S., Ghez C. and Eidelberg D. (2003) Impaired sequence learning in carriers of the DYT1 dystonia mutation. Ann. Neurol. 54, 102109.
  • Ichinose H, Ohye T., Takahashi E. et al. (1994) Hereditary progressive dystonia with marked diurnal fluctuation caused by mutations in the GTP cyclohydrolase I gene. Nature. Genet. 8, 236242.
  • Jankovic J. and Fahn S. (1993) Dystonic disorders, in Parkinson’s Disease and Movement Disorders (JankovicJ. and TolosaE., eds), pp. 337374. Williams and Wilkins, Baltimore.
  • Klien C., Ozelius L. J. and Breakefield X. O. (2007) Genetic evaluation in primary dystonia, in Handbook of Dystonia (StacyM. ed.), pp. 21–44. Informa Healthcare, New York.
  • Knappskog P. M., Flatmark T., Mallet J., Ludecke B. and Bartholome K. (1995) Recessively inherited L-DOPA-responsive dystonia caused by a point mutation (Q381 K) in the tyrosine hydroxylase gene. Hum. Mol. Genet. 4, 12091212.
  • Kuczenski R., Segal D. S. and Todd P. K. (1997) Behavioral sensitization and extracellular dopamine responses to amphetamine after various treatments. Psychopharmacology 134, 221229.
  • Leung J. C., Klein C., Friedman J. et al. (2001) Novel mutation in the TOR1A (DYT1) gene in atypical early onset dystonia and polymorphisms in dystonia and early onset parkinsonism. Neurogenetics 3, 133143.
  • Lupas A., Flagan J. M., Tamura T. and Baumeister W. (1997) Self-compartmentalization proteases. Trends Biochem. Sci. 22, 399404.
  • McNaught K. S., Kapustin A., Jackson T., Jengelley T. A., Jnobaptiste R., Shashidharan P., Perl D. P., Pasik P. and Olanow C. W. (2004) Brainstem pathology in DYT1 primary torsion dystonia. Ann. Neurol. 56, 540547.
  • Misbahuddin A., Placzek M. R., Taanman J. W., Gschmeissner S., Schiavo G., Cooper J. M. and Warner T. T. (2005) Mutant torsinA, which causes early-onset primary torsion dystonia, is redistributed to membranous structures enriched in vesicular monoamine transporter in cultured human SH-SY5Y cells. Mov. Dis. 20, 432440.
  • Mizoguchi K., Yuzurihara M., Nagata M., Ishige A., Sasaki H. and Tabira T. (2002) Dopamine-receptor stimulation in the prefrontal cortex ameliorates stress-induced rotarod impairment. Pharmacol. Biochem. Behav. 72, 723728.
  • Neuwald A. F., Aravind L., Spouge J. L. and Koonin E. V. (1999) AAA+: a class of chaperone-like ATPases associated with the assembly, operation, and disassembly of protein complexes. Genome Res. 9, 2743.
  • Ogura T. and Wilkinson A. J. (2001) AAA+ superfamily ATPases: common structure–diverse function. Genes Cells 6, 575597.
  • Ogura T., Ogata M., Akita H., Jitsuki S., Akiba L., Noda K., Hoka S. and Saji M. (2005) Impaired acquisition of skilled behavior in rotarod task by moderate depletion of striatal dopamine in a pre-symptomatic stage model of Parkinson’s disease. Neurosci. Res. 51, 299308.
  • Ozelius L. J., Hewett J., Page C. et al. (1997) The early-onset torsion dystonia gene (DYT1) encodes an ATP-binding protein. Nat. Genet. 17, 4048.
  • Rostasy K., Augood S. J., Hewett J. W., Leung J. C., Sasaki H., Ozelius L. J., Ramesh V., Standaert D. G., Breakefield X. O. and Hedreen J. C. (2003) TorsinA protein and neuropathology in early onset generalized dystonia with GAG deletion. Neurobiol. Dis. 12, 1124.
  • Rouge-Pont F., Usiello A., Benoit-Marand M., Gonon F., Piazza P. V. and Borrelli E. (2002) Changes in extracellular dopamine-induced by morphine and cocaine: crucial control by D2 receptors. J. Neurosci. 22, 32933301.
  • Rupniak N. M., Jenner P. and Marsden C. D. (1986) Acute dystonia induced by neuroleptic drugs. Psychopharmacology 88, 403419.
  • Sharma N., Baxter M. G., Petravicz J., Bragg C. D., Schienda A., Standaert D. G. and Breakefield X. O. (2005) Impaired motor learning in mice expressing torsinA with the DYT1 dystonia mutation. J. Neurosci. 25, 53515355.
  • Shashidharan P., Sandu D., Potla U., Armata I. A., Walker R. H., McNaught K. S., Weisz D., Sreenath T., Brin M. F. and Olanow C. W. (2005) Transgenic mouse model of early-onset DYT1 dystonia. Hum. Mol. Genetics 14, 125133.
  • Skidmore F. and Reich S. G. (2005) Tardive dystonia. Curr. Treat. Options Neurol. 7, 231236.
  • Sulzer D., Sonders M. S., Poulsen N. W. and Galli A. (2005) Mechanisms of neurotransmitter release by amphetamines: a review. Prog. Neurobiol. 75, 406433.
  • Torres G. E., Sweeney A. L., Beaulieu J. M., Shashidharan P. and Caron M. G. (2004) Effect of torsinA on membrane proteins reveals a loss of function and a dominant-negative phenotype of the dystonia-associated ΔE-torsinA mutant. Proc. Natl Acad. Sci. USA 101, 1565015655.
  • Vacher C. M., Gassmann M., Desrayaud S., Challet E., Bradaia A., Hoyer D., Waldmeier P., Kaupmann K., Pevet P. and Bettler B. (2006) Hyperdopaminergia and altered locomotor activity in GABAB1-deficient mice. J. Neurochem. 97, 979991.
  • Westerink B. H. C. (1995) Brain microdialysis and its application for the study of animal behaviour. Behav. Brain Res. 70, 103124.