Construction of a Mitochondria‐Endoplasmic Reticulum Dual‐Targeted Red‐Emitting Fluorescent Probe for Imaging Peroxynitrite in Living Cells and Zebrafish

Abstract

Peroxynitrite (ONOO−) is one of the important reactive oxygen species, which plays a vital role in the physiological process of intracellular redox balance. Revealing the biological functions of ONOO− will contribute to further understanding of the oxidative process of organisms. In this work, we designed and synthesized a novel red‐emitting fluorescent probe MCSA for the detection of ONOO−, which could rapidly respond to ONOO− within 250 s and exhibited high sensitivity to ONOO− with a low detection limit of 78 nM. Co‐localization experiments demonstrated MCSA had the ability to localize into the mitochondria and endoplasmic reticulum. What's more, MCSA enabled monitoring ONOO− level changes during tunicamycin‐induced endoplasmic reticulum stress. We have also successfully achieved the visual detection of exogenous and endogenous ONOO− in living cells and zebrafish. This work presented a chemical tool for imaging ONOO− in vitro and in vivo.

Recent Progress of Spectroscopic Probes for Peroxynitrite and Their Potential Medical Diagnostic Applications

“…With the use of the probe, the authors realized the ratiometric image visualization of the peroxynitrite in living cells. Very similar spectral behaviors were obtained with similar structure HDs Cy 11–15 [ 114 , 115 , 116 , 117 , 118 ], regardless of their difference in the quaternized heterocycles and aryl substituents.…”

“…The hemicyanine type of peroxynitrite probes could also be constructed via the condensation of the quaternized heterocycles with various aryl aldehydes. ( Figure 12 ) The aryl group in aryl aldehydes included EDG-substituted naphthalene ( Cy 12 [ 115 ] and Cy 13 [ 116 ]) and dihydronaphthalene ( Cy 14 [ 117 ]), coumarin ( Cy 15–20 [ 118 , 119 , 120 , 121 , 122 , 123 ]), porphrin ( Cy 21 [ 124 ]), and rhodamine ( Cy 22 [ 125 ])). The porphrin and rhodamine groups were relatively less electron-donating than others, leading to longer response times (90 min and 40 min, respectively).…”

Sensing Mechanism and Excited-State Dynamics of a Widely Used Intracellular Fluorescent pH Probe: pHrodo

“…Fluorescent probes act as great analytical tools for real-time monitoring of biological parameters and processes such as temperature, pH, ion concentration, and reactive oxygen species, which are invisible or inaccessible to the human eye. − Among them, the intracellular pH value, regulated by cell membrane proton pumps, is one of the most important to monitor because of its direct association with many pathological and physiological processes (e.g., enzyme activity, protein degradation, and mutations), the functions of organelles, and diseases (e.g., cancer, diabetes, and Alzheimer’s disease). − Hence, it is important to be able to detect intracellular pH in real time and improve our understanding of how these pH-mediated changes affect migration and metastasis processes of different cells as well as how they lead to diseases. , …”