Elevated cerebrospinal fluid ratios of cysteinyl-dopamine/3,4-dihydroxyphenylacetic acid in parkinsonian synucleinopathies.

INTRODUCTION: There is intense interest in identifying cerebrospinal fluid (CSF) biomarkers of Parkinson's disease (PD), both for early diagnosis and to track effects of putative treatments. Nigrostriatal dopamine depletion characterizes PD. Predictably, CSF levels of 3,4-dihydroxyphenylacetic acid (DOPAC), the main neuronal metabolite of dopamine, are decreased in PD, even in patients with recent onset of the movement disorder. Whether low CSF DOPAC is associated specifically with parkinsonism has been unclear. In the neuronal cytoplasm dopamine undergoes not only enzymatic oxidation to form DOPAC but also spontaneous oxidation to form 5-S-cysteinyl-dopamine (Cys-DA). Theoretically, oxidative stress or decreased activity of aldehyde dehydrogenase (ALDH) in the residual nigrostriatal dopaminergic neurons would increase CSF Cys-DA levels with respect to DOPAC levels. PD, parkinsonian multiple system atrophy (MSA-P), and pure autonomic failure (PAF) are synucleinopathies; however, PAF does not entail parkinsonism. We examined whether an elevated Cys-DA/DOPAC ratio provides a specific biomarker of parkinsonism in synucleinopathy patients. METHODS: CSF catechols were assayed in PD (n = 24), MSA-P (n = 32), PAF (n = 18), and control subjects (n = 32). RESULTS: Compared to controls, CSF DOPAC was decreased in PD and MSA-P (p < 0.0001 each). In both diseases Cys-DA/DOPAC ratios averaged more than twice control (0.14 ± 0.02 and 0.13 ± 0.02 vs. 0.05 ± 0.01, p < 0.0001 each), whereas in PAF the mean Cys-DA/DOPAC ratio was normal (0.05 ± 0.01). CONCLUSIONS: CSF Cys-DA/DOPAC ratios are substantially increased in PD and MSA-P and are normal in PAF. Thus, in synucleinopathies an elevated CSF Cys-DA/DOPAC ratio seems to provide a specific biomarker of parkinsonism.

Central dopamine deficiency in pure autonomic failure.

OBJECTIVE: Pure autonomic failure (PAF) and Parkinson's disease (PD) share several clinical laboratory abnormalities; however, PAF is not associated with parkinsonism. In this study, we tested the hypothesis that preservation of nigrostriatal dopaminergic innervation explains the absence of motor dysfunction in PAF. MMETHODS: Patients with PAF (N = 5) or PD (N = 21) and control subjects (N= 14) had brain 6-[18F]fluorodopa positron emission tomographic scanning and cerebrospinal fluid catechol measurements. A patient with PAF and another with PD had rapid postmortem striatal, nigral, and sympathetic ganglion sampling, with assays of catechols and tyrosine hydroxylase activity. RESULTS: The PAF and PD groups had similarly low mean substantia nigra (SN):occipital (OCC) ratios of 6-[18F]fluorodopa-derived radioactivity and similarly low cerebrospinal fluid dihydroxyphenylacetic acid and DOPA levels. Only the PD group, however, had low PUT:OCC, caudate:OCC, or PUT:SN ratios. The PAF and PD cases had similarly low SN tissue concentrations of dopamine and tyrosine hydroxylase activity, but the PD patient had tenfold lower PUT dopamine and the PAF patient 15-fold lower myocardial norepinephrine concentrations. CONCLUSIONS: Surprisingly, PAF and PD entail similarly severe nigral and overall central dopaminergic denervation. There is more severe loss of striatal dopaminergic terminals in PD than in PAF and more severe loss of sympathetic noradrenergic terminals in PAF than in PD. These differences explain the distinctive clinical manifestations of the two Lewy body diseases. Parkinsonism appears to reflect striatal dopamine deficiency rather than loss of nigral dopaminergic neurons per se.

Cerebrospinal fluid levels of catechols in patients with neurogenic orthostatic hypotension.

In multiple system atrophy (MSA) and pure autonomic failure (PAF), orthostatic hypotension (OH) results from deficient noradrenaline release from sympathetic nerves during standing. Post-mortem findings have indicated loss of central noradrenergic cells in both diseases. The present study sought in vivo neurochemical evidence for central noradrenergic deficiency in patients with OH due to MSA or PAF. A total of 28 patients with OH (18 with MSA; 10 with PAF) had cerebrospinal fluid and blood sampled for levels of noradrenaline and its neuronal metabolite dihydroxyphenylglycol. A control group of 44 subjects included 10 elderly normal volunteers, 10 patients with Alzheimer's disease, 18 patients with dysautonomia (postural tachycardia syndrome or neurocardiogenic syncope) and six patients with MSA in the absence of OH. Patients with OH had lower cerebrospinal fluid concentrations of noradrenaline (0.53+/-0.07 nmol/l) and dihydroxyphenylglycol (6.52+/-0.46 nmol/l) than did control subjects (0.90+/-0.09 and 9.64+/-0.46 nmol/l respectively; P =0.0001). The MSA+OH group had higher plasma levels of both catechols (noradrenaline, 1.31+/-0.16 nmol/l; dihydroxyphenylglycol, 5.08+/-0.43 nmol/l) than did the PAF group (noradrenaline, 0.38+/-0.08 nmol/l; dihydroxyphenylglycol, 2.53+/-0.30 nmol/l; P <0.001), despite similarly low cerebrospinal fluid levels. Among MSA patients, those with OH had lower cerebrospinal fluid levels of noradrenaline and dihydroxyphenylglycol than those without OH (noradrenaline, 1.71+/-0.64 nmol/l; dihydroxyphenylglycol, 10.41+/-1.77 nmol/l respectively; P =0.006). The findings are consistent with central noradrenergic deficiency in both MSA+OH and PAF. In MSA, central noradrenergic deficiency seems to relate specifically to OH.

Patterns of cerebrospinal fluid catechols support increased central noradrenergic responsiveness in aging and Alzheimer's disease.

BACKGROUND: High cerebrospinal fluid (CSF) norepinephrine (NE) concentrations in aging and Alzheimer's disease (AD) could reflect decreased NE clearance from central nervous system (CNS) extracellular fluid or increased release of NE into CNS extracellular fluid. Measuring CSF concentrations of the intraneuronal NE metabolite dihydroxyphenylglycol (DHPG), an estimate of NE clearance, and the NE precursor dihydroxyphenylacetic acid (DOPA), an estimate of NE biosynthesis, can help differentiate these mechanisms. METHODS: NE, DHPG, and DOPA were determined by HPLC in CSF and plasma obtained following yohimbine, clonidine, and placebo. Ten AD, 10 older, and 11 young subjects were studied. RESULTS: CSF DOPA following yohimbine was higher in older and AD than in young subjects. CSF DHPG did not differ among groups. Plasma DOPA following yohimbine was higher in AD than in young subjects. CONCLUSIONS: During alpha-2 adrenoreceptor blockade in both aging and AD, there are increased responses of CNS NE biosynthesis and release with unchanged CNS NE clearance. This pattern is consistent with partial loss of CNS noradrenergic neurons with compensatory activation of remaining CNS noradrenergic neurons. Given the marked loss of locus coeruleus (LC) noradrenergic neurons in AD, achievement of high CSF NE suggests particularly prominent compensatory activation of remaining LC neurons in this disorder.

Detection of salicylate and its hydroxylated adducts 2,3- and 2,5-dihydroxybenzoic acids as possible indices for in vivo hydroxyl radical formation in combination with catechol- and indoleamines and their metabolites in cerebrospinal fluid and brain tissue.

It has been suggested that salicylate (SA) hydroxylation can be used to detect hydroxyl radical formation in vivo. Here we describe a rapid and sensitive HPLC method using ultraviolet absorbance (UV) and electrochemical detection (EC) to detect SA (UV), its hydroxylated adducts 2,3- and 2,5-dihydroxybenzoic acids (DHBA) and catechol in combination with catechol- and indoleamines and related metabolites (EC) in one isocratic run. These compounds were measured in acidified cerebrospinal fluid (CSF) and perchlorate extracts of striatal tissues of untreated and SA-loaded rats (300 mg/kg SA, i.p.). Peaks were identified by comparing retention times of samples and standards, by adding standards to biological samples, by voltamograms, and by comparing chromatograms of manganese (Mn2+)-injected striata of SA-loaded rats with several control conditions. Six hours after unilateral injection of 0.4 mumol Mn2+ into striatum, 2,3-DHBA and 2,5-DHBA levels in striatum were respectively 4- and 7-fold increased as compared to non-injected (contralateral) striata, suggesting in vivo hydroxyl radical formation. In addition, dopamine and serotonin levels were depleted in Mn(2+)-injected striata by 46% and 64%, respectively. In CSF of Mn(2+)-injected rats, DHBA/SA ratios were not significantly changed as compared to those of control rats. In conclusion, the described technique can be applied to study in vivo hydroxyl radical formation in direct relation with dopaminergic and serotonergic neurotransmitter changes during neurotoxic processes.

Plasma and cerebrospinal fluid neurochemical pattern in Menkes disease.

Menkes disease is a neurodegenerative disorder of copper metabolism. Because the enzyme dopamine-beta-hydroxylase requires copper to catalyze the conversion of dopamine to norepinephrine, we reasoned that patients with Menkes disease would have a neurochemical pattern similar to that seen in patients with congenital absence of dopamine-beta-hydroxylase, i.e., high levels of dopamine, the dopamine metabolite dihydroxyphenylacetic acid (DOPAC), and the catecholamine precursor dihydroxyphenylalanine (DOPA), and low levels of norepinephrine and its neuronal metabolite dihydroxyphenylglycol (DHPG). We measured plasma and cerebrospinal fluid (CSF) levels of catechols in 10 patients ranging in age from 9 days to 27 months. In contrast to patients with congenital absence of dopamine-beta-hydroxylase, norepinephrine levels were normal in plasma of 4 Menkes patients and in CSF of all 10 patients. However, the ratios of DOPA:DHPG and DOPAC:DHPG in plasma and CSF of Menkes patients were invariably increased beyond the ranges of control values. These neurochemical findings indicate partial deficiency of dopamine-beta-hydroxylase in Menkes patients, with compensatory increases in catecholamine biosynthesis in sympathetic nerves and in the brain. Increased tyrosine hydroxylation and increased exocytotic release of norepinephrine may be responsible for preservation of plasma and CSF norepinephrine levels in Menkes patients. The abnormal neurochemical pattern, including high ratios of DOPA:DHPG and DOPAC:DHPG, may serve as a biochemical marker for Menkes disease and provide a baseline against which the influence of proposed therapies can be judged.

[Effects of thyrotropin-releasing hormone on neurologic recovery and biogenic amine metabolites of cerebrospinal fluid in spinal disorders].

The pharmacological effects of the thyrotropin-releasing hormone (TRH) in relation to biogenic amine metabolism in cerebrospinal fluid were examined in 35 patients with various spinal disorders. Neurologic conditions before and after TRH treatment were evaluated using subjective symptoms and Frankel's classification. Biogenic amine metabolism in cerebrospinal fluid was examined before and after TRH treatment measuring the metabolites by high performance liquid chromatography with electrochemical detection. Significant decreases in metabolites of norepinephrine and dopamine were seen in most cases of spinal disorders. The amount of serotonin metabolite, however, was not changed. In many acute cases, the neurologic condition was improved, and a significant increase in the dopamine metabolite was seen in the improved cases after TRH treatment. In chronic cases, TRH treatment was not as effective as in acute cases. TRH was therefore thought to be an effective agent in the treatment of acute spinal disorders. When an increase in the dopamine metabolite is seen after TRH treatment, neurologic improvement would probably be expected.

Abnormal central monoamine metabolism in humans with "true hypersomnia" and "sub-wakefulness".

A case of Kleine-Levin syndrome with true hypersomnia and a case of sub-wakefulness are described. In both patients lumbar cerebrospinal fluid homovanillic acid, 5-hydroxyindoleacetic acid, 3-methoxy-4-hydroxyphenylethylene glycol levels have been assayed during episodes of hypersomnia and normal sleep-waking cycles. Besides an increased 5-hydroxytryptamine turnover, mainly an increased dopamine turnover has been detected in both kinds of hypersomnia, and this finding was more remarkable in the case with sub-wakefulness. The probable role of dopamine in abnormalities in the sleep-waking cycle is discussed on the basis of results in experimental animal hypersomnias.

The effect of probenecid on the free and conjugaed 3-methoxy-4-hydroxyphenylglycol (MHPG) in lumbar cerebrospinal fluid.

Free and conjugated lumbar cerebrospinal fluid 3-methoxy-4-hydroxyphenylglycol (MHPG) was measured before and after probenecid treatment in 12 schizophrenic patients by a gas liquid chromatography-mass fragmentographic procedure. Neither the free nor conjugated MHPG was appreciably altered by probenecid. Total MHPG was statistically increased by probenecid but not to the point that the probenecid test would be clinically useful for estimating norepinephrine turnover from probenecid-induced changes in MHPG concentrations.

Central norepinephrine metabolism in affective illness: MHPG in the cerebrospinal fluid.

Concentrations of the norepinephrine metabolite 3-methoxy-4-hydroxyphenyl glycol in cerebrospinal fluid were measured by a gas chromatographic method in 34 patients with affective illness and in 44 controls. Concentrations of this metabolite in spinal fluid were significantly lower in depressed patients than in controls or manic patients. These low values may occur secondary to depressive phenomena such as reduced psychomotor activity, or they may reflect a primary change in norepinephrine metabolism in depressive illness.

Amine metabolites in human cerebrospinal fluid: effects of cord transection and spinal fluid block.

Patients with spinal cord transection had normal concentrations of 5-hydroxyindoleacetic acid and low concentrations of 3-methoxy-4-hydroxyphenyl glycol in lumbar cerebrospinal fluid. The presence or absence of spinal fluid block in these patients did not affect concentrations of either amine metabolite. However, the concentration of homovanillic acid was lower in patients with spinal fluid block than in those without block. The results suggest that the spinal cord contributes to concentrations of 3-methoxy-4-hydroxyphenyl glycol and possibly 5-hydroxyindoleacetic acid, but contributes little to that of homovanillic acid in the lumbar spinal fluid of man.