Construction of a Dual-Functional, Reversible, Ratiometric, Golgi-Targeting Fluorescent Probe for Real-time Monitoring of Dynamics of Intracellular Redox Homeostasis

Abstract

Intracellular redox homeostasis is highly important for the physiological processes of living organisms. Real-time monitoring of the dynamics of this intracellular redox process is pivotal but challenging because the biological redox reactions involved in the process are reversible and require at least one pair of oxidizing and reducing species. Thus, biosensors for investigating intracellular redox homeostasis need to be dual-functional, reversible, and, ideally, ratiometric in order for them to have real-time monitoring capacity and to provide accurate imaging information. In light of the importance of the redox pair between ClO– and GSH in living organisms, herein, we used the phenoselenazine (PSeZ) moiety as an electron donor and a reaction site to design a coumarin-based fluorescent probe, PSeZ-Cou-Golgi. After successive treatment with ClO– and GSH, the probe PSeZ-Cou-Golgi experienced an oxidation of selenium (Se) to selenoxide (SeO) by ClO– and a subsequent reduction of SeO to Se by GSH. The redox reactions alternatively changed the electron-donating strength of the donor in the probe PSeZ-Cou-Golgi, in turn affecting the intramolecular charge transfer process that resulted in the reversible, ratiometric change of fluorescence from red to green. After four cycles of reversible ClO–/GSH detection during in vitro experiments, the probe PSeZ-Cou-Golgi still had good performance. With the Golgi-targeting group, the probe PSeZ-Cou-Golgi was able to monitor the dynamic change of the ClO–/GSH-mediated redox state during Golgi oxidative stress, making it a versatile molecular tool. More importantly, the probe PSeZ-Cou-Golgi could facilitate the imaging of the dynamic redox state during acute lung injury progression.

Design, synthesis, and biological application of A–D–A-type boranil fluorescent dyes

“…26,27 However, excessive HClO can damage normal cells and tissues, and subsequently cause a variety of diseases such as cardiovascular disease, lung injury, neurodegenerative disease, atherosclerosis and even cancers. 31–35 The sulfur atom in organic compounds can be selectively oxidized into sulfoxide by HClO, which has been widely exploited as a strategy for the construction of HClO fluorescent probes. 36…”

Fluorescent probes for targeting the Golgi apparatus: design strategies and applications

“…In 2023, the Song group developed a dual-functional, reversible, and ratiometric fluorescent probe 21 with coumarin fluorophore, benzenesulfonamide for Golgi-targeting, and selenium (Se) as the recognition site. 75 Probe 21 could evaluate dynamic changes of Golgi oxidative stress by monitoring dynamic changes in the ClO − /GSH-mediated redox state in HeLa cells with rapid response, high selectivity, high sensitivity and low detection limit. More importantly, the probe facilitates the monitoring of the progression acute lung injury through the imaging of the dynamic redox state (Fig.…”

Tri-mode Responses to Reactive Oxygen Species In Vivo by Chiral Vanadium-Based Nanoparticles

“…However, there is a paucity of high-sensitivity biological sensors because the associated preparation conditions are often complex, costly, and potentially toxic and may suffer from stability issues. Traditional techniques for monitoring ROS, such as electron paramagnetic resonance, fluorescence, − chemiluminescence, , and electrochemical analysis, − typically have several limitations, including poor stability, low sensitivity, and cumbersome operation . Therefore, it is of utmost importance to develop new methods with high sensitivity and superior biocompatibility to measure ROS in complex biological environments.…”