Kojima H, Sakurai K, Kikuchi K, Kawahara S, Kirino Y, Nagoshi H, Hirata Y, Akaike T, Maeda H, Nagano T
Graduate School of Pharmaceutical Sciences, the University of Tokyo, Japan.
Biol Pharm Bull. 1997 Dec;20(12):1229-32. doi: 10.1248/bpb.20.1229.
Nitric oxide (NO) has been reported to play various roles as a signal transmitter. However, detailed functions of NO have yet to be clarified. We have developed a fluorescent indicator for NO imaging in living cells. The N-nitrosation of newly designed and synthesized 4-((3-amino-2-naphthyl)aminomethyl)benzoic acid (DAN-1) by NO yielded the highly fluorescent triazole-form. The membrane permeable ester derivative of DAN-1 (DAN-1 EE) was applied to the imaging of NO produced in activated rat aortic smooth muscle cells. After DAN-1 EE has been loaded into cells, the ester bond is hydrolyzed by intracellular esterase, yielding original DAN-1 with less permeability. The fluorescence intensity of the cells loaded with DAN-1 EE increased according to NO production. The imaging method with fluorescent indicators will be significant for the functional clarification of NO in vivo.
据报道，一氧化氮（NO）作为信号传递者发挥着多种作用。然而，NO的详细功能尚未阐明。我们开发了一种用于活细胞中NO成像的荧光指示剂。新设计合成的4-((3-氨基-2-萘基)氨基甲基)苯甲酸（DAN-1）被NO进行N-亚硝化反应后，生成了高荧光的三唑形式。DAN-1的膜可渗透酯衍生物（DAN-1 EE）被应用于活化的大鼠主动脉平滑肌细胞中产生的NO的成像。在DAN-1 EE被加载到细胞中后，酯键被细胞内酯酶水解，产生通透性较低的原始DAN-1。加载了DAN-1 EE的细胞的荧光强度根据NO的产生而增加。使用荧光指示剂的成像方法对于体内NO功能的阐明将具有重要意义。
