Juanita L. Carl

Dynamic Changes in Striatal Dopamine D2 and D3 Receptor Protein and mRNA in Response to 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine (MPTP) Denervation in Baboons

Loss of nigrostriatal neurons leads to striatal dopamine deficiency and subsequent development of parkinsonism. The effects of this denervation on D2-like receptors in striatum remain unclear. Most studies have demonstrated increases in striatal dopamine D2-like receptors in response to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-mediated denervation, but others have found either decreases or no change in binding. To clarify the response to denervation, we have investigated the time-dependent changes in dopamine D2, D3, and D4 receptor protein and mRNA levels in unilaterally MPTP-lesioned baboons. MPTP (0.4 mg/kg) was infused into one internal carotid artery, producing a contralateral hemi-parkinsonian syndrome. After MPTP treatment, the animals were maintained for 17–480 d and then euthanized. MPTP decreased ipsilateral dopamine content by >90%, which did not change with time. Ipsilateral D2-like receptor binding in caudate and putamen initially decreased then increased two- to sevenfold over the first 100 d and returned to near baseline levels by 480 d. Relative levels of D2mRNA were essentially unchanged over this period. D4 mRNA was not detected. In contrast, D3 mRNA increased sixfold by 2 weeks and then decreased. At the peak period of increase in binding sites, all D2-like receptors were in a micromolar affinity agonist-binding state, implying an increase in uncoupled D2but not D3 receptor protein. Taken together, these data suggest that MPTP-induced changes in D2-like dopamine receptors are complex and include translational or post-translational mechanisms.

Long term treatment and disease severity change brain responses to levodopa in Parkinson’s disease

Objectives: Degeneration of nigrostriatal neurons and subsequent striatal dopamine deficiency produce many of the symptoms of Parkinson disease (PD). Initially restoration of striatal dopamine with oral levodopa provides substantial benefit, but with long term treatment and disease progression, levodopa can elicit additional clinical symptoms, reflecting altered effects of levodopa in the brain. The authors examined whether long term treatment affects the brain’s response to levodopa in the absence of these altered clinical responses to levodopa. Methods: Positron emission tomography (PET) measurements were used of brain-blood flow before and after an acute dose of levodopa in three groups: PD patients treated long term with levodopa without levodopa induced dyskinesias, levodopa naive PD patients, and controls. Results: It was found that the PD group treated long term responded to acute levodopa differently from controls in left sensorimotor and left ventrolateral prefrontal cortex. In both regions, the treated PD group had decreased blood flow whereas the control group had increased blood flow in response to levodopa. Levodopa naive PD patients had little or no response to levodopa in these regions. Within the treated PD group, severity of parkinsonism correlated with the degree of abnormality of the sensorimotor cortex response, but not with the prefrontal response. Conclusions: It is concluded that long term levodopa treatment and disease severity affect the physiology of dopaminergic pathways, producing altered responses to levodopa in brain regions associated with motor function.

Levodopa Challenge Neuroimaging of Levodopa-Related Mood Fluctuations in Parkinson's Disease

Some patients with advanced Parkinsons disease (PD) develop dose-related fluctuations in mood. This may reflect alterations in dopamine-influenced brain circuits that mediate emotion. However, there is no available information to localize which dopamine-influenced neurons may be most affected. Eight patients with PD and clinically significant levodopa-related mood fluctuations (mania, depression, or anxiety) were compared to 13 patients with similarly severe PD and fluctuations of motor function but not of mood. Regional cerebral blood flow (rCBF) was measured with positron emission tomography before and after levodopa (in the presence of carbidopa). The rCBF response to levodopa in medial frontal gyrus and posterior cingulate cortex (PCC) significantly differed between mood fluctuators and control patients (corrected p<0.02). Other regions with uncorrected p<0.001 in this comparison were cortical Brodmann areas 22, 40, 13, 11, and 28, hippocampus, and claustrum. The levodopa activation paradigm detected group differences not evident in a comparison of resting rCBF. Abnormalities of dopamine innervation may produce mood fluctuations via effects on PCC, an area strongly linked to mood and anxiety and with known rCBF responsiveness to levodopa or D2-like dopamine receptor agonists. We speculate that mood fluctuations may arise in parkinsonian patients who have abnormal dopaminergic modulation of caudate nucleus, anterior cingulate cortex, or orbital frontal cortex, all of which innervate PCC. The findings require confirmation in larger and better-matched groups.

Dopa-Induced Blood Flow Responses in Nonhuman Primates

Initially, treatment with the dopamine precursor levodopa provides substantial symptomatic relief for patients with Parkinsons disease (PD). However, as the disease progresses, side effects such as involuntary movements or psychosis may accompany the response to medication. The mechanisms underlying these actions of levodopa remain unclear. To develop methodology for longitudinal studies of the effects of PD and levodopa treatment in living nonhuman primates, we first studied the effects of an acute dose of levodopa on regional brain activity in sedated baboons using positron emission tomography. We found that levodopa significantly decreased regional cerebral blood flow (rCBF) bilaterally in putamen and right cingulate and increased rCBF in right lateral temporal cortex and bilateral frontal cortex. We then performed similar studies on a nemestrina in awake and sedated states to determine whether these responses were affected by sedation. Interestingly, the directions of the rCBF responses in the putamen and temporal cortex were reversed depending on the presence or absence of sedation. Specifically, responses were decreased in sedated animals, but increased dose-dependently in the awake nemestrina. These findings have important implications for the interpretation of studies that use anesthesia. The responses in the awake nemestrina were most similar to those reported in humans and thus may be the most useful model system. Future imaging studies using selective dopaminergic agents in awake animals may permit the identification of relatively specific agonist-mediated pathways and may help separate the mechanisms that mediate levodopas benefit from those that produce its unwanted side effects.

Rapid intravenous loading of levodopa for human research: clinical results

Levodopa has several advantages as a pharmacological challenge agent for human neuroscience research. Exogenous levodopa changes striatal neuronal activity and increases extracellular dopamine concentrations, and with adequate inhibition of peripheral metabolism levodopa does not change mean cerebral blood flow. For neuroimaging studies of Parkinson disease (PD) and Tourette syndrome, we sought to rapidly produce a biologically relevant steady-state levodopa concentration and then maintain that concentration for at least an hour. We also wished to minimize side effects, even in individuals without prior levodopa treatment. We designed a two-stage intravenous infusion protocol based on published levodopa pharmacokinetic data. We report results of 125 infusions in 106 subjects, including healthy volunteers, PD patients, and people with chronic tics. At higher doses (target steady-state levodopa concentrations of 2,169 and 1,200 ng/ml), treatment-naive volunteers had unacceptably frequent side effects. The final infusion protocol, with a target steady-state concentration of 600 ng/ml, was well-tolerated (mild nausea in 11% of subjects was the only side effect occurring significantly more than in single-blind saline infusions), produced the desired plasma levodopa concentration (612+/-187 ng/ml, mean+/-S.D.), and produced statistically significant antiparkinsonian benefit (16% mean reduction in a standard rating of parkinsonian motor signs, P<0.0005).

Carbohydrate metabolism in brain during convulsions and its modification by phenobarbitone.

Abstract— Assays of citric acid cycle substrates and metabolites of the second stage of the glycolytic pathway have completed a series of studies of glucose metabolism in brains of mice rapidly frozen at intervals during electrically‐induced, tonic‐clonic convulsions. Citric acid cycle metabolism reached a new equilibrium at a significantly higher rate. However, oxidative metabolism did not keep up with the demand for energy supplies, as indicated by an increasing lactate level and an increasing lactate: pyruvate ratio. Administration of a sub‐anaesthetic but anticonvulsant dose of phenobarbitone prior to convulsive electrical stirnulation was associated with as great an increase in anaerobic glycolysis as in mice given no drug prior to stimulation; but oxidative metabolism was not enhanced, as reflected by even greater lactate: pyruvate ratios in mice given phenobarbitone than in mice given no drug prior to convulsive stimulation.

Cognitive-pharmacologic functional magnetic resonance imaging in tourette syndrome: a pilot study

BACKGROUND Dopamine agonists and antagonists can reduce abnormal movements and vocalizations (tics) in Tourette syndrome (TS); however, dopamine-responsive abnormal function in specific brain regions has not been directly demonstrated in TS. We sought to identify dopamine-modulated brain regions that function abnormally in TS by combining functional magnetic resonance imaging (fMRI), a working memory (WM) task, and infusion of the dopamine prodrug levodopa (while blocking dopamine production outside the brain). METHODS We obtained complete fMRI data in 8 neuroleptic-naive adults with a chronic tic disorder and in 10 well-matched tic-free control subjects. RESULTS Different task-sensitive brain regions responded differently to the WM task depending on levodopa status and diagnostic group (analysis of variance [ANOVA], p <.001). Four regions showed interactions with diagnosis (ANOVA, p <.001). In TS subjects, the task induced excessive brain activity in parietal cortex, medial frontal gyrus, and thalamus. Levodopa normalized the excess activity. In left parietal cortex, the degree of normalization was greater in patients with higher levodopa plasma concentrations (n = 6; Spearmans r = -.84, p =.04) and a greater degree of diagnostic confidence of TS (r = -.71, p =.05). CONCLUSIONS These results are consistent with a dopamine-influenced functional abnormality of brain response in TS and suggest testable hypotheses about the mechanism by which dopamine antagonists and agonists alleviate tics.

Effects of duration of convulsions on energy reserves of the brain.

Abstract— Rapid administration (0·4 ml in 1 sec) of the convulsant inhalant, flurothyl (hexafluorodiethyl ether, indoklon), to mice induced within 10 sec clonic‐tonic seizures that were accompanied by marked decrease of P‐creatine, decrease of ATP and glucose, and increase of lactate in the cerebral cortex. In contrast, mice to which flurothyl had been administered slowly (0·05 ml every 30 sec for 10 min) exhibited myoclonic jerks after about 3 min, merging into irregular clonus at about 5 min, and intermittent clonus thereafter until onset of tonic hind limb extension at about 10 min. In these mice, P‐creatine in cerebral cortex decreased gradually for 6 min and then remained through 10 min at levels nearly as low as those reached 10 sec after rapidly administered flurothyl. Lactate in cerebral cortex increased much more during the 10 min of slow administration of flurothyl than in 10 sec of rapid administration, the greatest increase occurring at 4–6 min, as myoclonic jerks merged into irregular clonus. Glucose and ATP in cerebral cortex fluctuated somewhat but did not decrease greatly when flurothyl was administered slowly.

Cocaine and Amphetamine Modification of Cerebral Energy Metabolism in vivo

At the time of maximal behavioral stimulation after injection of amphetamine (5 mg/kg) in mice, elevation of cerebral cortical levels of malate in the citric acid cycle and of the amino acid, alanine, was observed, suggesting that this drug has widespread effects on energy metabolism. Cocaine (20 mg/kg), in contrast, produced elevation of brain glucose but not of citric acid cycle substrates or amino acids at the time of maximal hyperactivity. These observations are discussed in terms of the mechanisms of action of these two central nervous system stimulants.

Brain amino acids during convulsions.

A RAPID fall in brain a-ketoglutarate before the onset of clinical seizures was demonstrated in a study of citric acid cycle metabolites after convulsive electrical stimulation in mice (KING et al., 1973). During recovery from convulsions, citric and cycle metabolism reached a new equilibrium at a significantly higher rate. Prior administration of phenobarbitone prevented the changes in a-ketoglutarate and in citric acid cycle metabolism in general. These observations prompted the studies described in this paper of glutamate, glutamine and GABA. all of which are closely related to the metabolism of a-ketoglutarate. and of aspartate and alanine, the other two amino acids most closely associated to citric acid cycle metabolites or precursors (oxalacetate and pyruvate, respectively).