A microassay for measuring synaptosomal super(3)H-dopamine and super(3)H-metabolite release

A microassay for measuring synaptosomal super(3)H-dopamine and super(3)H-metabolite release

A modified synaptosomal superfusion apparatus is described which uses less than 10 micrograms of tissue per replicate sample and facilitates the routine separation of tritiated dopamine ( super(3)H-DA), super(3)H-dihydroxypheny-acetic acid ( super(3)H-DOPAC), and super(3)H-homovanillic acid ( super(...

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Veröffentlicht in:Brain research bulletin 1990-01, Vol.25 (3), p.423-427
Hauptverfasser: Petrie, E C, Lombrozo, L, Csernansky, J G
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Lombrozo, L
Csernansky, J G
description A modified synaptosomal superfusion apparatus is described which uses less than 10 micrograms of tissue per replicate sample and facilitates the routine separation of tritiated dopamine ( super(3)H-DA), super(3)H-dihydroxypheny-acetic acid ( super(3)H-DOPAC), and super(3)H-homovanillic acid ( super(3)H-HVA). A flow rate of 1.5 ml/min allows superfusion without the use of reuptake or monoamine oxidase inhibitors. Superfusate samples are collected directly onto alumina columns for the separation of super(3)H-DA and its acid metabolites. Total recovery of authentic super(3)H-DA applied via superfusion was 87.63(1.10) percent (Mean(SEM)). Contamination of the acetic acid eluate fraction, containing 80.98(1.15)% of the total DA, by DOPAC and HVA was less than 0.1%. To illustrate the utility of this technique, the relative proportions of super(3)H-DA and super(3)H-metabolites released from synaptosomes by 6 mM potassium potassium and 1 mu M reserpine were compared.
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title A microassay for measuring synaptosomal super(3)H-dopamine and super(3)H-metabolite release
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