A New Technique for Simultaneous Assay of Biogenic Amines and Their Metabolites in Unpurified Mouse Brain

Morier, E.; Rips, Richard

doi:10.1080/01483918208068827

Cited by 46 publications

(7 citation statements)

References 5 publications

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“…Regional monoamine distribution agreed with previous results for rats (Glowinski and Iversen, 1966) and mice (Morier and Rips, 1982): cerebellum, medulla-pons, frontal cortex, and hippocampus contained 5-HT and noradrenaline with 20, 39, 52, and 46% of 5-HT, respectively (G. Boschi, unpublished data). Hypothalamus and striatum contained 5-HT, dopamine, and noradrenaline in the respective ratios of 28:11:61 and 3:95:2.…”

Section: Discussionsupporting

confidence: 90%

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Dihydrotetrabenazine Binding and Monoamine Uptake in Mouse Brain Regions

Scherman

1986

Journal of Neurochemistry

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Binding of [2-3H]dihydrotetrabenazine and uptake of 5-hydroxytryptamine (serotonin) were studied in mouse brain cerebellum, pons-medulla, frontal cortex, hypothalamus, hippocampus, and striatum. Binding of [2-3H]dihydrotetrabenazine to homogenates of these brain areas is stable for several hours and occurs at a homogeneous class of binding sites ( K D = 2.4 nM). Subcellular fractionation and regional distribution of [2-3H]dihydrotetrabenazine binding and serotonin uptake showed that the ligand binds to synaptic vesicles. Dihydrotetrabenazine inhibited serotonin uptake with the same inhibitory constant (IC50 = 2.6 nM) for synaptic vesicles from brain regions containing 3,4-dihydroxyphenylethylamine (dopamine) or serotonin and noradrenaline in different proportions. This constant is similar to the K D of [2-3H]dihydrotetrabenazine, which suggests that the latter ligand labels specifically and with the same affinity the monoamine transporter from various mono-aminergic synaptic vesicles. Therefore the regional differences in central monoamine depletion induced in vivo by tetrabenazine are not due to regional differences in inhibition of vesicular monoamine uptake. Moreover, vesicular monoamine transporters from the central and peripheral nervous systems of various mammals and from bovine adrenal glands have comparable affinity for substrate and inhibitor ( K , values for serotonin and ICso for dihydrotetrabenazine are about 0.8 p M and 3 nM, respectively) and comparable turnover number (10-35 molecules transported per transporter per minute), which suggests the involvement of a common transporter molecule in the process of monoamine uptake by the various monoaminergic storage vesicles. Key Words: Monoamine uptake-Synaptic vesi~les-[2-~H]Dihydrotetrabenazine binding-Tetrabenazine. Scherman D. Dihydrotetrabenazine binding and monoamine uptake in mouse brain regions. 5-HT, 5-hydroxytryptamine (serotonin); TBZ, tetrabenazine; Pierre and Marie Curie, Pans 75005, France. TBZOH, dihydrotetrabenazine. Whittaker V. P., Michaelson I. A., and Kirkland R. J. (1963) The separation of synaptic vesicles from disrupted nerve-ending particles. Biochem. Pharmacol. 12, 300-302. 694-706.

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Section: Discussionsupporting

confidence: 90%

“…Protein concentration was measured by the method of Lowry et al (1951). Catecholamines and 5-HT were mea- sured as described by Morier and Rips (1982). For phospholipid determination, samples were hydrolyzed for I h at 180°C in perchloric acid (70%) and inorganic phosphate measured as described by Lowry and Lopez (1946).…”

Section: Miscellaneousmentioning

confidence: 99%

Dihydrotetrabenazine Binding and Monoamine Uptake in Mouse Brain Regions

Scherman

1986

Journal of Neurochemistry

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“…Quantitation of DA, S-HT, and their metabolites DA, S-HT, DOPAC, HVA, and 5-HIAA were quantitated by a modification of the HPLC technique of Morier and Rips (1982) in aliquots taken from samples bcing prcpared for quantitation of ACh and Ch. The HPLC system consisted of a Model M45 solvent delivery system connected to a WISP 710B automatic sample injector from Waters Associates (Milford, MA, U.S.A.), which had been modified with the addition of a refrigeration unit (Lauda Model RM6) to maintain a temperature of 7°C.…”

Section: Determination Of Ach Turnovermentioning

confidence: 99%

Effect of Intrastriatal Injection of Diisopropylfluorophosphate on Acetylcholine, Dopamine, and Serotonin Metabolism

Robinson

Rice

Hambrecht

1986

Journal of Neurochemistry

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Diisopropylfluorophosphate (81.5 nmol) was injected directly into the striata of rats to study changes in striatal metabolism of acetylcholine (ACh), 3,4-dihydroxyphenylethylamine (dopamine), and 5-hydroxytryptamine (serotonin) at early time points following acute irreversible inhibition of cholinesterase. Twenty minutes following the intrastriatal injection of diisopropylfluorophosphate, levels of striatal acetylcholine were elevated by 50%, but a decrease in KACh compensated for this change. At 1 h, levels of ACh were still elevated, but not significantly different from control values. However, KACh and, hence, ACh turnover were greatly enhanced at this time. Finally, at 24 h, striatal ACh content was only slightly elevated and KACh and the turnover rate of ACh had returned to control values. Striatal cholinesterase activity remained significantly inhibited at all three times. At none of these times was ACh content or turnover affected in the parietal cortex, hippocampus, hypothalamus, or medulla/pons. Neither dopamine and its metabolites 3,4-dihydroxyphenylacetic acid and homovanillic acid nor serotonin and its metabolite 5-hydroxyindoleacetic acid were significantly affected at any of the three times by intrastriatal diisopropylfluorophosphate treatment. Possible mechanisms of the changes in cholinergic parameters are discussed.

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“…Analytical procedure A Waters 6000 A liquid chromatograph with a U 6 K injector (Waters Associates, Milford, U.S.A.), a Waters RCM-100 radial compression system, a Waters Radial Pak A column (C 18, 0-8 cm i.d. x 10 cm, 10 ,sm particle size), a Metrohm 641 electrochemical detector fitted with two vitrous carbon electrodes EA 286-1, and one reference Ag/AgCl EA 442 electrode were used according to the technique described by Morier & Rips (1982). The potential used was +0-8 V. The mobile phase consisted of 0.1 M-KH2PO4 (Merck), 0-1 mM-EDTA (Prolabo), 0-8 mM-heptane sulphonic acid (Waters Pic B7) in a mixture of demineralized water (Crouzat, > 5 x 10-6 Q cm2), and methanol (85:15).…”

Section: Animals and Operative Proceduresmentioning

confidence: 99%

On the significance of monoamines and their metabolites in the cerebrospinal fluid of the sheep.

Ruckebusch¹,

Sutra²

1984

The Journal of Physiology

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SUMMARY1. Assays capable of concurrently measuring small quantities of noradrenaline, dopamine, serotonin, and several of their metabolites in cerebrospinal fluid (c.s.f.) were developed by the use of high performance liquid chromatography with electrochemical detection.2. For comparison, cortical subarachnoid, ventricular, cisternal and lumbar c.s.f. were obtained by puncture under barbiturate anaesthesia in sheep. Basal concentrations related to the adrenergic system, including methoxyhydroxyphenylglycol (MHPG), were similar in ventricular, cisternal and lumbar c.s.f., and those of the serotoninergic metabolites, 5-hydroxyindole-3-acetylacetic acid (5-HIAA), were similar in ventricular and cisternal c.s.f. High concentrations of the dopamine metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were found only in ventricular c.s.f.3. Monoamine metabolites in ventricular c.s.f. under basal conditions and after various experimental manipulations were then determined over periods of 3 months in two different breeds of sheep fitted chronically with cannulae in lateral ventricles. A dose-related accumulation of all the acidic monoamine metabolites was recorded during treatment with probenecid. The increase in 5-HIAA was linear after administration of increased doses of tryptophan and 5-hydroxytryptophan. The concentrations of dopamine, DOPAC and HVA in the ventricular c.s.f. reflected the response of the dopaminergic system to agents capable of crossing the blood-brain barrier.4. It is concluded that cerebral metabolism in conscious sheep could be indirectly approached by recording the concentration of end-products of dopamine metabolism in ventricular cerebrospinal fluid, obtained under conditions of minimal stress.

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A New Technique for Simultaneous Assay of Biogenic Amines and Their Metabolites in Unpurified Mouse Brain

Cited by 46 publications

References 5 publications

Dihydrotetrabenazine Binding and Monoamine Uptake in Mouse Brain Regions

Dihydrotetrabenazine Binding and Monoamine Uptake in Mouse Brain Regions

Effect of Intrastriatal Injection of Diisopropylfluorophosphate on Acetylcholine, Dopamine, and Serotonin Metabolism

On the significance of monoamines and their metabolites in the cerebrospinal fluid of the sheep.

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