Abstract
The measurement of nitric oxide (NO) is important for direct examination of the regulatory roles of NO in various biological systems. Diaminofluoresceins (DAFs), new fluorescence indicators for NO, were applied to detect the release of NO from bovine aortic endothelial cells (ECs). DAFs react with NO to yield the corresponding green-fluorescent triazolofluoresceins, which provide the advantages of specificity, sensitivity and a simple protocol for the direct detection of NO. Using these DAFs, we could detect the generation of NO not only from inducible NO synthase expressed in macrophages, but also from constitutive NO synthase expressed in ECs.
MeSH terms
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Animals
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Aorta
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Cattle
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Cells, Cultured
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Endothelium, Vascular / metabolism*
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Enzyme Inhibitors / pharmacology
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Fluoresceins*
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Indicators and Reagents
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Macrophages / metabolism
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Mice
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Nitric Oxide / biosynthesis*
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Nitric Oxide Synthase / antagonists & inhibitors
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Nitric Oxide Synthase / metabolism
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Nitric Oxide Synthase Type II
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Nitric Oxide Synthase Type III
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Spectrometry, Fluorescence / methods*
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omega-N-Methylarginine / pharmacology
Substances
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4,5-diamino-2-(3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid
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5,6-diamino-2-(2,7-dichloro-3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid
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Enzyme Inhibitors
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Fluoresceins
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Indicators and Reagents
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omega-N-Methylarginine
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Nitric Oxide
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Nitric Oxide Synthase
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Nitric Oxide Synthase Type II
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Nitric Oxide Synthase Type III
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Nos2 protein, mouse
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Nos3 protein, mouse