Detection and Imaging of Nitric Oxide with Novel Fluorescent Indicators: Diaminofluoresceins
| Autor: | Kazuya Kikuchi, Yasunobu Hirata, Naoki Nakatsubo, Hirotatsu Kojima, Tetsuo Nagano, Yutaka Kirino, Hiroshi Nagoshi, Shigenori Kawahara |
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| Rok vydání: | 1998 |
| Předmět: |
Male
Cell Membrane Permeability Triazole Aorta Thoracic Nitric Oxide Muscle Smooth Vascular Analytical Chemistry Nitric oxide chemistry.chemical_compound In vivo Microscopy Animals Reactivity (chemistry) Rats Wistar Cells Cultured Detection limit Microscopy Confocal fungi Reproducibility of Results Fluorescence Culture Media Rats Spectrometry Fluorescence chemistry Biochemistry Biophysics Fluorescein Indicators and Reagents Visible spectrum |
| Zdroj: | Analytical Chemistry. 70:2446-2453 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac9801723 |
| Popis: | Nitric oxide is a gaseous, free radical which plays a role as an intracellular second messenger and a diffusable intercellular messenger. To obtain direct evidence for NO functions in vivo, we have designed and synthesized diaminofluoresceins (DAFs) as novel fluorescent indicators for NO. The fluorescent chemical transformation of DAFs is based on the reactivity of the aromatic vicinal diamines with NO in the presence of dioxygen. The N-nitrosation of DAFs, yielding the highly green-fluorescent triazole form, offers the advantages of specificity, sensitivity, and a simple protocol for the direct detection of NO (detection limit 5 nM). The fluorescence quantum efficiencies are increased more than 100 times after the transformation of DAFs by NO. Fluorescence detection with visible light excitation and high sensitivity enabled the practical assay of NO production in living cells. Membrane-permeable DAF-2 diacetate (DAF-2 DA) can be used for real-time bioimaging of NO with fine temporal and spatial resolution. The dye 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. |
| Databáze: | OpenAIRE |
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