Near-infrared rhodol dyes bearing salicylaldehyde moieties for ratiometric pH sensing in live cells during mitophagy and under hypoxia conditions

Dwivedi, Sushil K.; Arachchige, Dilka Liyana; Vohs, Tara; Tang, Jiayou; Usimaki, Kyle; Olowolagba, Adenike; Fritz, Delaney Raine; Luck, Rudy L.; Werner, Thomas; Liu, Haiying

doi:10.1039/d2tb02791g

Cited by 10 publications

(7 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This consumption establishes a controlled baseline, optimal for scrutinizing NAD(P)H fluctuations in response to various treatments. 13,55 In our investigations, control larvae treated with probe C in PBS exhibited minimal fluorescence, indicating consumption of NAD(P)H (Figure 14). Upon exposure to different concentrations of NADH, we observed a dose-dependent rise in fluorescence, mirroring the increase in NAD(P)H levels (Figure 14).…”

Section: Visualizing Dynamic Cellular Nad(p)h Fluctuations Across Var...mentioning

confidence: 64%

“…We employed freshly starved-hatched fruit fly larvae as a model system to assess the efficacy of probe C in monitoring NAD(P)H dynamics in vivo. 13,55 The selection of these larvae was due to their reduced NAD(P)H levels, a result of metabolic adjustments in response to lacking nutrients upon hatching. 13,55 In the absence of food, larval metabolism consumes NAD(P)H, a crucial coenzyme central to cellular reactions.…”

Section: Visualizing Dynamic Cellular Nad(p)h Fluctuations Across Var...mentioning

confidence: 99%

“…13,55 The selection of these larvae was due to their reduced NAD(P)H levels, a result of metabolic adjustments in response to lacking nutrients upon hatching. 13,55 In the absence of food, larval metabolism consumes NAD(P)H, a crucial coenzyme central to cellular reactions. This consumption establishes a controlled baseline, optimal for scrutinizing NAD(P)H fluctuations in response to various treatments.…”

Section: Visualizing Dynamic Cellular Nad(p)h Fluctuations Across Var...mentioning

confidence: 99%

See 2 more Smart Citations

Syntheses and Applications of Coumarin-Derived Fluorescent Probes for Real-Time Monitoring of NAD(P)H Dynamics in Living Cells across Diverse Chemical Environments

Olowolagba,

Idowu,

Arachchige

et al. 2024

ACS Appl. Bio Mater.

View full text Add to dashboard Cite

Fluorescent probes play a crucial role in elucidating cellular processes, with NAD(P)H sensing being pivotal in understanding cellular metabolism and redox biology. Here, the development and characterization of three fluorescent probes, A, B, and C, based on the coumarin platform for monitoring of NAD(P)H levels in living cells are described. Probes A and B incorporate a coumarin-cyanine hybrid structure with vinyl and thiophene connection bridges to 3-quinolinium acceptors, respectively, while probe C introduces a dicyano moiety for replacement of the lactone carbonyl group of probe A which increases the reaction rate of the probe with NAD(P)H. Initially, all probes exhibit subdued fluorescence due to intramolecular charge transfer (ICT) quenching. However, upon hydride transfer by NAD(P)H, fluorescence activation is triggered through enhanced ICT. Theoretical calculations confirm that the electronic absorption changes upon the addition of hydride to originate from the quinoline moiety instead of the coumarin section and end up in the middle section, illustrating how the addition of hydride affects the nature of this absorption. Control and dose−response experiments provide conclusive evidence of probe C's specificity and reliability in identifying intracellular NAD(P)H levels within HeLa cells. Furthermore, colocalization studies indicate probe C's selective targeting of mitochondria. Investigation into metabolic substrates reveals the influence of glucose, maltose, pyruvate, lactate, acesulfame potassium, and aspartame on NAD(P)H levels, shedding light on cellular responses to nutrient availability and artificial sweeteners. Additionally, we explore the consequence of oxaliplatin on cellular NAD(P)H levels, revealing complex interplays between DNA damage repair, metabolic reprogramming, and enzyme activities. In vivo studies utilizing starved fruit fly larvae underscore probe C's efficacy in monitoring NAD(P)H dynamics in response to external compounds. These findings highlight probe C's utility as a versatile tool for investigating NAD(P)H signaling pathways in biomedical research contexts, offering insights into cellular metabolism, stress responses, and disease mechanisms.

show abstract

Section: Visualizing Dynamic Cellular Nad(p)h Fluctuations Across Var...mentioning

confidence: 64%

Section: Visualizing Dynamic Cellular Nad(p)h Fluctuations Across Var...mentioning

confidence: 99%

Section: Visualizing Dynamic Cellular Nad(p)h Fluctuations Across Var...mentioning

confidence: 99%

See 1 more Smart Citation

Syntheses and Applications of Coumarin-Derived Fluorescent Probes for Real-Time Monitoring of NAD(P)H Dynamics in Living Cells across Diverse Chemical Environments

Olowolagba,

Idowu,

Arachchige

et al. 2024

ACS Appl. Bio Mater.

View full text Add to dashboard Cite

show abstract

“…An MTT assay was utilized to verify the toxicity of the probes, as described previously. ,– A549 cells were subjected to either probe A or B , and the enzymatic reduction of the yellow MTT dye to a purple formazan product in the mitochondria was measured as an indicator of cell viability. The MTT assay shows that cell viability is above 80% under a concentration of 40 μM (Figures S18 and S19), indicating that the probes are of low toxicity to the cells.…”

Section: Results and Discussionmentioning

confidence: 99%

Highly Sensitive Cyanine Dyes for Rapid Sensing of NAD(P)H in Mitochondria and First-Instar Larvae of Drosophila melanogaster

Arachchige

Dwivedi

Jaeger

et al. 2023

ACS Appl. Bio Mater.

Self Cite

View full text Add to dashboard Cite

We have developed two highly sensitive cyanine dyes, which we refer to as probes A and B. These dyes are capable of quick and sensitive sensing of NAD(P)H. The dyes were fabricated by connecting benzothiazolium and 2,3dimethylnaphtho[1,2-d]thiazol-3-ium units to 3-quinolinium through a vinyl bond. In the absence of NAD(P)H, both probes have low fluorescence and absorption peaks at 370 and 400 nm, correspondingly. This is because of their two electron-withdrawing acceptor systems with high charge densities. However, when NAD(P)H reduces the probes' electron-withdrawing 3-quinolinium units to electron-donating 1,4-dihydroquinoline units, the probes absorb at 533 and 535 nm and fluoresce at 572 and 586 nm for A and B correspondingly. This creates well-defined donor−π− acceptor cyanine dyes. We successfully used probe A to monitor NAD(P)H levels in live cells during glycolysis, under hypoxic conditions induced by CoCl 2 treatment and after treatment with cancer drugs, including cisplatin, camptothecin, and gemcitabine. Probe A was also employed to visualize NAD(P)H in Drosophila melanogaster first-instar larvae. We observed an increase in NAD(P) H levels in A549 cancer cells both under hypoxic conditions and after treatment with cancer drugs, including cisplatin, camptothecin, and gemcitabine.

show abstract

“…S15, ESI †). 26 The results indicated that both probes exhibited excellent biocompatibility, with cell viability maintained at 90% even at a high probe concentration of 50 mM (Fig. S15, ESI †).…”

Section: Probe Cytotoxicity and Photostabilitymentioning

confidence: 96%

Thiophene-based organic dye with large Stokes shift and deep red emission for live cell NAD(P)H detection under varying chemical stimuli

Dwivedi,

Arachchige,

Olowolagba

et al. 2023

J. Mater. Chem. B

Self Cite

View full text Add to dashboard Cite

show abstract

Near-infrared rhodol dyes bearing salicylaldehyde moieties for ratiometric pH sensing in live cells during mitophagy and under hypoxia conditions

Abstract: We describe a simple but efficient approach to make fluorescent probes A and B based on rhodol dyes incorporated with salicyaldehyde moiety for monitoring pH changes in mitochondria under oxidative...

Cited by 10 publications

References 42 publications

Syntheses and Applications of Coumarin-Derived Fluorescent Probes for Real-Time Monitoring of NAD(P)H Dynamics in Living Cells across Diverse Chemical Environments

Syntheses and Applications of Coumarin-Derived Fluorescent Probes for Real-Time Monitoring of NAD(P)H Dynamics in Living Cells across Diverse Chemical Environments

Highly Sensitive Cyanine Dyes for Rapid Sensing of NAD(P)H in Mitochondria and First-Instar Larvae of Drosophila melanogaster

Thiophene-based organic dye with large Stokes shift and deep red emission for live cell NAD(P)H detection under varying chemical stimuli

Contact Info

Product

Resources

About