Single Fluorescent Probe Responds to H₂O₂, NO, and H₂O₂/NO with Three Different Sets of Fluorescence Signals

Abstract

Hydrogen peroxide (H(2)O(2)) acts as a signaling molecule in a wide variety of signaling transduction processes and an oxidative stress marker in aging and disease. However, excessive H(2)O(2) production is implicated with various diseases. Nitric oxide (NO) serves as a secondary messenger inducing vascular smooth muscle relaxation. However, mis-regulation of NO production is associated with various disorders. To disentangle the complicated inter-relationship between H(2)O(2) and NO in the signal transduction and oxidative pathways, fluorescent reporters that are able to display distinct signals to H(2)O(2), NO, and H(2)O(2)/NO are highly valuable. Herein, we present the rational design, synthesis, spectral properties, and living cell imaging studies of FP-H(2)O(2)-NO, the first single-fluorescent molecule, that can respond to H(2)O(2), NO, and H(2)O(2)/NO with three different sets of fluorescence signals. FP-H(2)O(2)-NO senses H(2)O(2), NO, and H(2)O(2)/NO with a fluorescence signal pattern of blue-black-black, black-black-red, and black-red-red, respectively. Significantly, we have further demonstrated that FP-H(2)O(2)-NO, a single fluorescent probe, is capable of simultaneously monitoring endogenously produced NO and H(2)O(2) in living macrophage cells in multicolor imaging. We envision that FP-H(2)O(2)-NO will be a unique molecular tool to investigate the interplaying roles of H(2)O(2) and NO in the complex interaction networks of the signal transduction and oxidative pathways. In addition, this work establishes a robust strategy for monitoring the multiple ROS and RNS species (H(2)O(2), NO, and H(2)O(2)/NO) using a single fluorescent probe, and the modularity of the strategy may allow it to be extended for other types of biomolecules.

Effects of Single and Double Bonds in Linkers on Colorimetric and Fluorescent Sensing Properties of Polyving Akohol Grafting Rhodamine Hydrazides

“…The fluorescence turn-off were further supported by the observation that the emission color of the probe solution turned from pale pink to deep purple for Cu 2+ , and from pale pink with orange red to orange for Fe 3+ , respectively (Fig. 4) [22,31,44,45].…”

“…As seen from Fig. 7, there were small changes of fluorescence intensity for PVA-SR6GH adding Ag + , Ba 2+ , Cd 2+ , Co 2+ , Fe 2+ , K + , La 3+ , Mg 2+ , Na + , Ni 2+ , Pb 2+ , and Zn 2+ cations, but there were highest fluorescence enhancement for PVA-SR6GH upon addition of Cu 2+ , Fe 3+ , Cr 3+ and Hg 2+ [30,31,44,46,47].…”

“…It could be seen that the fluorescence signal of the PVA-MRBH with Cu 2+ ion remarkably decreased for a few minutes, and leveled off as the time continues. The fluorescence intensity of PVA-MRBH with Cu 2+ reached its minimum value at about 10 min, after which the fluorescence intensity remained almost constant [26,[40][41][42][43][44].…”

“…The relative fluorescence intensity enhanced nearly 1.64, 1.55, 1.60, or 4.14 times, respectively. However, we attempted addition of Cu 2+ , Fe 3+ , Cr 3+ or Hg 2+ ions to aqueous solution of the PVA-SR6GH and did not find any significant changes in the relative fluorescence intensity [18,22,31,44,45]. Figure S12 showed the comparative response of PVA-SR6GH to Cu 2+ , Fe 3+ , Cr 3+ or Hg 2+ in aqueous solution.…”

Light-Up of Rhodamine Hydrazide to Generate Emissive Initiator for Polymerization and to Afford Photochromic Polypeptide Metal Complex

“…Non-emissive, transparent R-C [33,34] (Scheme 3) became emissive and purple in color at the instant PLG-NCA [35] was added. Rhodamine is able to respond to external stimuli, such as pH, to undergo the reversible closed rhodamine spirolactam into opened amide transformation [36][37][38][39][40]. Accompanied with the structural change is the photophysical properties change; that is, the switching ON and OFF of the absorption band at about 550 nm, as well as the luminescence at about 580 nm.…”