Abstract
A xanthene derivative, Granada Green dinitrobenzene sulfonate (GGDNBS), has been synthesized to assay cellular oxidative stress based on changes in the concentration of biothiols. The dye is able to react with biological thiols by a thiolysis reaction that promotes a change in fluorescence intensity. To demonstrate the usefulness of GGDNBS for in vivo oxidative stress measurements, 661 W photoreceptor-derived cells were exposed to light to induce ROS generation, and changes in GGDNBS fluorescence were measured. In these cells, GGDNBS fluorescence was correlated with the biothiol levels measured by an enzymatic method. Therefore, GGDNBS allows us to monitor changes in the levels of biothiols associated with ROS generation via single-cell bioimaging.
Highlights
New methods to measure biothiols are continuously being developed[4,5]
We have recently explored the photophysical properties of Granada Green (GG), a xanthenic structure developed in our lab, and its derivatives for the in vivo detection of different analytes, including biothiols[33]
We describe a new use of a known group, 2,4-dinitrobenzene sulfonate (DNBS)[7,12,34,35], in Granada Green (GG) to obtain the derivative Granada Green dinitrobenzene sulfonate (GGDNBS) (Fig. 1)
Summary
New methods to measure biothiols are continuously being developed[4,5]. Among them, fluorescence-based approaches are the most interesting ones considering their advantages derived from their high sensitivity, simplicity and low cost[6]. Considerable effort has been made to develop new biothiol probes using this strategy, including detecting biothiols in serum and live cells through ratiometric measurements[14], simultaneously detecting biothiols and phosphate[15], and using ex vivo methods to detect intraperitoneal tumor nodules[16] or fluorophores with large Stokes shift[17]. Other successful biothiol intracellular probes using different mechanisms of actions, such as a reversible fluorescent biothiol probe[18], selective detection of GSH over other biothiols such as cysteine or homocysteine in cells[19], selenocysteine[20] or selective detection of thiophenols[21,22] have been reported. The development of reversible and ratiometric probes have been demonstrated very useful in the determination of intracellular biothiols[18,30,31] including inside mitochondria[32] we report here the use of a “turn-on” probe
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