David D. Song

Differential neuropathological alterations in transgenic mice expressing α-synuclein from the platelet-derived growth factor and Thy-1 promoters

Accumulation of α‐synuclein has been associated with neurodegenerative disorders, such as Lewy body disease and multiple system atrophy. We previously showed that expression of wild‐type human α‐synuclein in transgenic mice results in motor and dopaminergic deficits associated with inclusion formation. To determine whether different levels of human α‐synuclein expression from distinct promoters might result in neuropathology mimicking other synucleopathies, we compared patterns of human α‐synuclein accumulation in the brains of transgenic mice expressing this molecule from the murine Thy‐1 and platelet‐derived growth factor (PDGF) promoters. In murine Thy‐1‐human α‐synuclein transgenic mice, this protein accumulated in synapses and neurons throughout the brain, including the thalamus, basal ganglia, substantia nigra, and brainstem. Expression of human α‐synuclein from the PDGF promoter resulted in accumulation in synapses of the neocortex, limbic system, and olfactory regions as well as formation of inclusion bodies in neurons in deeper layers of the neocortex. Furthermore, one of the intermediate expresser lines (line M) displayed human α‐synuclein expression in glial cells mimicking some features of multiple system atrophy. These results show a more widespread accumulation of human α‐synuclein in transgenic mouse brains. Taken together, these studies support the contention that human α‐synuclein expression in transgenic mice might mimic some neuropathological alterations observed in Lewy body disease and other synucleopathies, such as multiple system atrophy.

Enhanced substantia nigra mitochondrial pathology in human alpha-synuclein transgenic mice after treatment with MPTP.

Recent studies have implicated alpha-synuclein (alpha-S) in the pathogenesis of Parkinsons disease (PD). The mechanisms underlying PD are not completely understood; however, mitochondrial complex I inhibition and oxidative injury may be involved. Because the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a potent complex I inhibitor that can cause oxidative injury and mimic many aspects of PD in treated animals, we sought to determine whether the overexpression of alpha-S in transgenic (tg) mice (alpha-S-tg) would enhance the substantia nigra (SN) pathology resulting from treatment with MPTP. For this purpose, alpha-S-tg mice were produced expressing high levels of wild-type (wt) human alpha-S under the control of the neuron-specific Thy-1 promoter. Alpha-S-tg mice and non-tg controls were treated with MPTP (15 mg/kg ip, twice a week for 2 weeks) or saline (Sal) and then examined 2 weeks after completion of treatment by transmission electron microscopy (EM). We found that alpha-S-tg mice treated with MPTP had extensive mitochondrial alterations, increases in mitochondrial size, filamentous neuritic aggregations, axonal degeneration, and formation of electron dense perinuclear cytoplasmic inclusions in the SN that did not occur in the hippocampus or neocortex, nor in MPTP-treated non-tg mice or Sal-treated alpha-S-tg mice. These findings support the potential involvement of alpha-S expression in the vulnerability of SN neurons to toxicity from mitochondrial complex I inhibitors and the subsequent development of neurodegenerative pathology.

Pilot trial of high dosages of coenzyme Q10 in patients with Parkinson's disease

The safety and tolerability of high dosages of coenzyme Q10 were studied in 17 patients with Parkinsons disease (PD) in an open label study. The subjects received an escalating dosage of coenzyme Q10--1200, 1800, 2400, and 3000 mg/day with a stable dosage of vitamin E (alpha-tocopherol) 1200 IU/day. The plasma level of coenzyme Q10 was measured at each dosage. Thirteen of the subjects achieved the maximal dosage, and adverse events were typically considered to be unrelated to coenzyme Q10. The plasma level reached a plateau at the 2400 mg/day dosage and did not increase further at the 3000 mg/day dosage. Our data suggest that in future studies of coenzyme Q10 in PD, a dosage of 2400 mg/day (with vitamin E/alpha-tocopherol 1200 IU/day) is an appropriate highest dosage to be studied.

Neurological and Neurodegenerative Alterations in a Transgenic Mouse Model Expressing Human α-Synuclein under Oligodendrocyte Promoter: Implications for Multiple System Atrophy

Multiple system atrophy (MSA) is a progressive, neurodegenerative disease characterized by parkinsonism, ataxia, autonomic dysfunction, and accumulation of α-synuclein (α-syn) in oligodendrocytes. To better understand the mechanisms of neurodegeneration and the role of α-syn accumulation in oligodendrocytes in the pathogenesis of MSA, we generated transgenic mouse lines expressing human (h) α-syn under the control of the murine myelin basic protein promoter. Transgenic mice expressing high levels of hα-syn displayed severe neurological alterations and died prematurely at 6 months of age. Furthermore, mice developed progressive accumulation of hα-syn-immunoreactive inclusions in oligodendrocytes along the axonal tracts in the brainstem, basal ganglia, cerebellum, corpus callosum, and neocortex. The inclusions also reacted with antibodies against phospho-serine (129) hα-syn and ubiquitin, and hα-syn was found in the detergent-insoluble fraction. In high-expresser lines, the white matter tracts displayed intense astrogliosis, myelin pallor, and decreased neurofilament immunostaining. Accumulation of hα-syn in oligodendrocytes also leads to prominent neurodegenerative changes in the neocortex with decreased dendritic density and to loss of dopaminergic fibers in the basal ganglia. The oligodendrocytic inclusions were composed of fibrils and accompanied by mitochondrial alterations and disruption of the myelin lamina in the axons. Together, these studies support the contention that accumulation of α-syn in oligodendrocytes promotes neurodegeneration and recapitulates several of the key functional and neuropathological features of MSA.

Information-Integration Category Learning in Patients With Striatal Dysfunction

Information-integration category learning was examined in patients with Parkinsons disease (PD) and in healthy control participants in 2 different conditions. In the linear condition, optimal categorization required a nonverbalizable linear integration of information from the 2 stimulus dimensions, whereas in the nonlinear condition, a nonlinear integration of information was required. Each participant completed 600 trials in each condition and was given corrective feedback following each trial. Results indicated that PD patients were not impaired in the linear condition across all trials, whereas the same patients were impaired in the nonlinear condition, but only later in training. The authors conducted model-based analyses to identify participants who used an information-integration approach, and a comparison of the accuracy rates of those individuals further revealed a specific deficit in information-integration category learning in patients with PD. These findings suggest that the striatum may be particularly involved in information-integration category learning when the rule is highly complex.

Neurobehavioral mechanisms of temporal processing deficits in Parkinson's disease.

Background Parkinsons disease (PD) disrupts temporal processing, but the neuronal sources of deficits and their response to dopamine (DA) therapy are not understood. Though the striatum and DA transmission are thought to be essential for timekeeping, potential working memory (WM) and executive problems could also disrupt timing. Methodology/Findings The present study addressed these issues by testing controls and PD volunteers ‘on’ and ‘off’ DA therapy as they underwent fMRI while performing a time-perception task. To distinguish systems associated with abnormalities in temporal and non-temporal processes, we separated brain activity during encoding and decision-making phases of a trial. Whereas both phases involved timekeeping, the encoding and decision phases emphasized WM and executive processes, respectively. The methods enabled exploration of both the amplitude and temporal dynamics of neural activity. First, we found that time-perception deficits were associated with striatal, cortical, and cerebellar dysfunction. Unlike studies of timed movement, our results could not be attributed to traditional roles of the striatum and cerebellum in movement. Second, for the first time we identified temporal and non-temporal sources of impaired time perception. Striatal dysfunction was found during both phases consistent with its role in timekeeping. Activation was also abnormal in a WM network (middle-frontal and parietal cortex, lateral cerebellum) during encoding and a network that modulates executive and memory functions (parahippocampus, posterior cingulate) during decision making. Third, hypoactivation typified neuronal dysfunction in PD, but was sometimes characterized by abnormal temporal dynamics (e.g., lagged, prolonged) that were not due to longer response times. Finally, DA therapy did not alleviate timing deficits. Conclusions/Significance Our findings indicate that impaired timing in PD arises from nigrostriatal and mesocortical dysfunction in systems that mediate temporal and non-temporal control-processes. However, time perception impairments were not improved by DA treatment, likely due to inadequate restoration of neuronal activity and perhaps corticostriatal effective-connectivity.

Microglial response is poorly correlated with neurodegeneration following chronic, low-dose MPTP administration in monkeys.

Many investigators have reported extensive microglial activation in the mouse substantia nigra and striatum following acute, high-dose 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration. Our previous work demonstrated tyrosine hydroxylase (TH)-positive fiber sprouting in the striatum in monkeys that had received a partial dopaminergic lesion using a low-dose, chronic MPTP administration paradigm. To characterize the microglial response, we utilized HLA-DR (LN3) to immunolabel the class II major histocompatibility complex (MHC II). In MPTP-treated monkeys, there was an intense microglial response in the substantia nigra, nigrostriatal tract, and in both segments of the globus pallidus. This response was morphologically heterogeneous, with commingled ramified, activated, and multicellular morphologies throughout the extent of these basal ganglia structures. Surprisingly, there was little evidence of microglial reactivity in the striatum despite evidence of neurodegeneration-by silver labeling and by loss of TH immunolabeling. Moreover, this pattern of microglial reactivity was the same in all animals that had received MPTP and seemed to be independent of the degree of neurotoxin-induced neurodegeneration. Thus, we conclude that microglial reactivity, per se, is not consistently associated with neurodegeneration, but depends on regional differences.

Spatial and object working memory deficits in Parkinson's disease are due to impairment in different underlying processes.

Working memory maintenance processes for visual-spatial and visual-object information were evaluated in patients with Parkinsons disease (PD). PD patients and controls performed a working memory task with two conditions that differed only in the aspect of the stimuli that the participant was instructed to remember: their locations or shapes. Maintenance processes were investigated by measuring accuracy over 1-s, 5-s, and 10-s delays. Results indicated that patients were impaired in maintaining object information over the delay. In contrast, the patients showed impairment on the spatial condition only when the to-be-remembered stimulus was highly similar in location to the probe, but this impairment was equivalent across the delays, suggesting that this deficit was not due to maintenance impairment. These results suggest that deficits in working memory for spatial and object information are mediated by distinct cognitive processes in nondemented patients with PD and may differ in their pathophysiological basis.

The impact of irrelevant dimensional variation on rule-based category learning in patients with Parkinson's disease

This study examined the impact of irrelevant dimensional variation on rule-based category learning in patients with Parkinsons disease (PD), older controls (OC), and younger controls (YC). Participants were presented with 4-dimensional, binary-valued stimuli and were asked to categorize each into 1 of 2 categories. Category membership was based on the value of a single dimension. Four experimental conditions were administered in which there were zero, 1, 2, or 3 randomly varying irrelevant dimensions. Results indicated that patients with PD were impacted to a greater extent than both the OC and YC participants when the number of randomly varying irrelevant dimensions increased. These results suggest that the degree of working memory and selective attention requirements of a categorization task will impact whether PD patients are impaired in rule-based category learning, and help to clarify recent discrepancies in the literature.

White-Matter Changes Correlate with Cognitive Functioning in Parkinson’s Disease

Diffusion tensor imaging (DTI) findings from emerging studies of cortical white-matter integrity in Parkinson’s disease (PD) without dementia are inconclusive. When white-matter changes have been found, their relationship to cognitive functioning in PD has not been carefully investigated. To better characterize changes in tissue diffusivity and to understand their functional significance, the present study conducted DTI in 25 PD patients without dementia and 26 controls of similar ages. An automated tract-based DTI method was used. Fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) were analyzed. Neuropsychological measures of executive functioning (working memory, verbal fluency, cognitive flexibility, inhibitory control) and visuospatial ability were then correlated with regions of interest that showed abnormal diffusivity in the PD group. We found widespread reductions in FA and increases in MD in the PD group relative to controls. These changes were predominantly related to an increase in RD. Increased AD in the PD group was limited to specific frontal tracks of the right hemisphere, possibly signifying more significant tissue changes. Motor symptom severity did not correlate with FA. However, different measures of executive functioning and visuospatial ability correlated with FA in different segments of tracts, which contain fiber pathways to cortical regions that are thought to support specific cognitive processes. The findings suggest that abnormal tissue diffusivity may be sensitive to subtle cognitive changes in PD, some of which may be prognostic of future cognitive decline.