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

Eric C. Petrie*, Leon Lombrozo, John G. Csernansky

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

A modified synaptosomal superfusion apparatus is described which uses less than 10 micrograms of tissue per replicate sample and facilitates the routine separation of 3H-DA, 3H-DOPAC, and 3H-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 3H-DA and its acid metabolites. Total recovery of authentic 3H-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 3H-DA and 3H-metabolites released from synaptosomes by 6 mM potassium and 1 μM reserpine were compared.

Original languageEnglish (US)
Pages (from-to)423-427
Number of pages5
JournalBrain Research Bulletin
Volume25
Issue number3
DOIs
StatePublished - Sep 1990

Keywords

  • Alumina
  • Chromatography
  • DOPAC
  • Dopamine
  • HVA
  • In vitro
  • Nucleus accumbens
  • Rat
  • Reserpine
  • Superfusion
  • Synaptosomes

ASJC Scopus subject areas

  • Neuroscience(all)

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