title: Development of a fluorescent indicator for nitric oxide based on the fluorescein chromophore.
abstract: Endogenous nitric oxide (NO) appears to modulate many physiological and pathophysiological processes. In order to obtain direct evidence for NO functions in vivo, we have developed 4,5-diaminofluorescein (DAF-2) as a novel fluorescent indicator for NO. Green-fluorescent triazolofluorescein formed by the reaction of NO and DAF-2 affords high sensitivity for NO (detection limit: 5 nM). Membrane-permeable DAF-2 diacetate (DAF-2 DA) was loaded into activated rat aortic smooth muscle cells, where the ester bonds are hydrolyzed by intracellular esterase, generating DAF-2. The fluorescence in the cells increased in a NO concentration-dependent manner. This imaging method should be useful for studies of the dynamic biological actions of NO at the molecular level with fine temporal and spatial resolution.
authors.affiliation: Graduate School of Pharmaceutical Sciences, University of Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kojima
authors.forename: K
authors.initials: K
authors.lastname: Sakurai
authors.forename: K
authors.initials: K
authors.lastname: Kikuchi
authors.forename: S
authors.initials: S
authors.lastname: Kawahara
authors.forename: Y
authors.initials: Y
authors.lastname: Kirino
authors.forename: H
authors.initials: H
authors.lastname: Nagoshi
authors.forename: Y
authors.initials: Y
authors.lastname: Hirata
authors.forename: T
authors.initials: T
authors.lastname: Nagano
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D019793:Fluorescein / Q000138:chemical synthesis* / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D009131:Muscle, Smooth, Vascular / Q000737:chemistry* / Q000378:metabolism; D009569:Nitric Oxide / Q000032:analysis*; D051381:Rats
chemical_list: C111205:4,5-diaminofluorescein; D005456:Fluorescent Dyes; D009569:Nitric Oxide; D019793:Fluorescein