Reliable in vitro measurement of nitric oxide released from endothelial cells using low concentrations of the fluorescent probe 4,5-diaminofluorescein

被引：90

作者：

Leikert, JF ^{[1
]}

Räthel, TR ^{[1
]}

Müller, C ^{[1
]}

Vollmar, AM ^{[1
]}

Dirsch, VM ^{[1
]}

机构：

[1] Univ Munich, Dept Pharm, Ctr Drug Res, D-81377 Munich, Germany

来源：

FEBS LETTERS | 2001年 / 506卷 / 02期

关键词：

nitric oxide; 4,5-diaminofluorescein; fluorescence spectroscopy; endothelial nitric oxide synthase;

D O I：

10.1016/S0014-5793(01)02901-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

4,5-Diaminofluorescein (DAF-2) and its membrane-permeable derivate DAF-2 diacetate are fluorescent probes that have been developed to perform real-time biological detection of nitric oxide (NO). Their use for intracellular imaging, however, has recently been seriously questioned and data using DAF-2 for extracellular NO detection at low levels, as for example released from endothelial cells, are rare. Here we show that a reliable detection of low levels of NO in biological systems by DAF-2 is possible (a) by using low DAF-2 concentrations (0.1 muM) and (b) by subtracting the DAF-2 auto-fluorescence from the measured total fluorescence. The described method allows easy real-time detection of endothelial NO formation. (C) 2001 Published by, Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

引用

页码：131 / 134

页数：4

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