Rational Design of a Targetable Fluorescent Probe for Visualizing H₂S Production under Golgi Stress Response Elicited by Monensin

Abstract

As a eukaryotic organelle, the Golgi apparatus plays an essential role in various physiological activities such as stress response. The Golgi stress response is an important physiological process of conferring cytoprotection by regulating the synthesis and metabolism of bioactive molecules. Therefore, the development of new suitable in situ analytical techniques for monitoring related small molecular substances in the stress reaction of the Golgi apparatus is very helpful for further study of the regulatory mechanism of the Golgi apparatus. Recent studies have shown that endogenous hydrogen sulfide (H2S) also possesses crucial bioregulatory and protective performances in the stress response. Therefore, the high-fidelity in situ mapping of H2S production under the Golgi stress response plays an important role not only in revealing cytoprotection functions of H2S in the stress response but also in further understanding the regulatory mechanism of the Golgi stress response. In this work, we designed a simple Golgi-targetable H2S fluorescent probe (Gol-H 2 S) that responds accurately and sensitively to H2S in the Golgi apparatus of living cells and zebrafish. On the basis of its superior bioimaging performances, probe Gol-H 2 S was successfully applied to the in situ visualization of H2S production under the Golgi stress response elicited by monensin, a specific-Golgi stressor. The related process of the Golgi stress response was validated by stimulation and inhibition experiments. These findings fully demonstrate that H2S is an alternative biomarker of the Golgi stress response. Moreover, probe Gol-H 2 S can also be used as a potential tool for disclosing the detailed H2S-cytoprotection mechanisms under the regulation of the Golgi stress response in related diseases.

An Activatable and Switchable Nanoaggregate Probe for Detecting H2S and Its Application in Mice Brains

“…We then applied this nanoprobe to monitor exogenous and endogenous H 2 S in HeLa cells (Figure 4). It was reported that exogenous Cys could stimulate cells to produce endogenous H 2 S in HeLa cells [48] . As we expected, the green and red fluorescence in group c was more obvious, compared with the control group a and b, which implied that HSDPP could be used to image endogenous and basal H 2 S in HeLa cells.…”

“…It was reported that exogenous Cys could stimulate cells to produce endogenous H 2 S in HeLa cells. [48] As we expected, the green and red fluorescence in group c was more obvious, compared with the control group a and b, which implied that HSDPP could be used to image endogenous and basal H 2 S in HeLa cells. On the contrary, N-ethylmaleimide (NEM) was incubated with HeLa cells to investigate that the nanoprobe HSDPP could selectively detect H 2 S in living cells.…”

BODIPY‐based Fluorescent Probe for Fast Detection of Hydrogen Sulfide and Lysosome‐targeting Applications in Living Cells

“…[28][29][30][31][32] Currently, most of the excellent researches for detecting H 2 S were the reaction-based methods that utilizing the high nucleophilicity and strong reduction capability of H 2 S, which are based on the following strategies such as Michael addition reaction, reduction of azide, nucleophilic reaction, thiolysis of dinitrophenyl ether, etc. [33][34][35][36][37][38][39][40][41][42][43][44][45][46][47][48][49] These advances have greatly promoted the research of H 2 S optical detection, However, fluorescent probes targeting hydrogen sulfide in lysosomes are still very rare, [50][51][52] so the construction of real-time, fast and specific fluorescent probe to detect H 2 S in lysosomes are important for understanding its molecular mechanism in life activities. BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) dyes have many excellent features, such as high molar absorption coefficients and fluorescence intensity, fine chemical and photo stability, pH insensitive nature.…”

“…Comparing with these methods, fluorescence techniques are more desirable due to its simplicity, high sensitivity and suitability for intracellular detection [28–32] . Currently, most of the excellent researches for detecting H 2 S were the reaction‐based methods that utilizing the high nucleophilicity and strong reduction capability of H 2 S, which are based on the following strategies such as Michael addition reaction, reduction of azide, nucleophilic reaction, thiolysis of dinitrophenyl ether, etc [33–49] . These advances have greatly promoted the research of H 2 S optical detection, However, fluorescent probes targeting hydrogen sulfide in lysosomes are still very rare, [50–52] so the construction of real‐time, fast and specific fluorescent probe to detect H 2 S in lysosomes are important for understanding its molecular mechanism in life activities.…”

A Golgi Apparatus-Targeting, Naphthalimide-Based Fluorescent Molecular Probe for the Selective Sensing of Formaldehyde

“…Hence, there is still a strong need for the development of an efficient probe that is highly specific for a particular sub-organelle and the development of FA-targeted fluorescent probes with a high specificity towards the Golgi body is of significant importance for revealing the biological processes associated with various diseases. Meanwhile, H 2 O 2 - and H 2 S-targeted fluorescent probes incorporated with the phenylsulfonamide moiety have been reported to stain the Golgi body, showing high Pearson coefficients over 0.92 [ 20 , 21 ].…”