Kojima H, Sakurai K, Kikuchi K, Kawahara S, Kirino Y, Nagoshi H, Hirata Y, Nagano T
Graduate School of Pharmaceutical Sciences, University of Tokyo, Japan.
Chem Pharm Bull (Tokyo). 1998 Feb;46(2):373-5. doi: 10.1248/cpb.46.373.
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.
内源性一氧化氮（NO）似乎可调节许多生理和病理生理过程。为了获得NO在体内功能的直接证据，我们开发了4,5-二氨基荧光素（DAF-2）作为一种新型的NO荧光指示剂。由NO与DAF-2反应形成的绿色荧光三唑荧光素对NO具有高灵敏度（检测限：5 nM）。将可透过细胞膜的DAF-2二乙酸酯（DAF-2 DA）载入活化的大鼠主动脉平滑肌细胞，在细胞内，酯键被细胞内酯酶水解，生成DAF-2。细胞内的荧光以NO浓度依赖的方式增加。这种成像方法对于在分子水平上以良好的时间和空间分辨率研究NO的动态生物学作用应该是有用的。
