Abstract

Intracellular concentrations of L-citrulline (Cit) and its metabolites are related to nitric oxide synthase (NOS) activity, an enzyme producing the intercellular messenger NO in animal tissues including the nervous system. A capillary electrophoresis system using laser-induced fluorescence detection is described, and methods are developed to monitor the levels of L-arginine (Arg), Cit, and related molecules in identified neurons of the marine slugs, Pleurobranchaea californica and Aplysia californica. The limits of detection for Arg, Cit, L-argininosuccinate, L-ornithine, and L-arginine phosphate range from 50 amol to 17 fmol (5 nM to 17 μM in the neurons under study); these detection limits are significantly lower than actual intracellular levels of the metabolites, allowing the direct assay of single cells. The levels of NOS metabolites in individual neurons varied from 6 (Arg) and 4 mM (Cit) in putative NOS-containing neurons down to <1 μM (undetectable) levels in many putative NOS-negative cells. The Arg/Cit ratio is independent of cell volume, correlates with NADPH-diaphorase staining, and appears to be a characteristic parameter for the presence of NOS activity in identified neurons.

Original languageEnglish (US)
Pages (from-to)2243-2247
Number of pages5
JournalAnalytical chemistry
Volume70
Issue number11
DOIs
StatePublished - Jun 1 1998

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Capillary electrophoresis
Metabolites
Nitric Oxide Synthase
Neurons
Arginine
NADPH Dehydrogenase
Citrulline
Ornithine
Enzyme activity
Neurology
Assays
Animals
Fluorescence
Tissue
Molecules
Lasers

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Capillary Electrophoresis Analysis of Nitric Oxide Synthase Related Metabolites in Single Identified Neurons. / Floyd, Philip D.; Moroz, Leonid L.; Gillette, Rhanor; Sweedler, Jonathan V.

In: Analytical chemistry, Vol. 70, No. 11, 01.06.1998, p. 2243-2247.

Research output: Contribution to journalArticle

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