Intracellular pH Measurements in Glioblastoma Cells Using the pH-Sensitive Dye BCECF

Galifianakis, Nataliya; Placantonakis, Dimitris G.; Chesler, Mitchell

doi:10.1007/978-1-4939-7659-1_8

Cited by 5 publications

(2 citation statements)

References 6 publications

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“…Measurement of pH i was performed according to the literature (Lee et al, 2007; Galifianakis et al, 2018). Fluorescent indicator BCECF/AM was used to monitor pH i changes.…”

Section: Methodsmentioning

confidence: 99%

Sulfur Dioxide Activates Cl-/HCO3- Exchanger via Sulphenylating AE2 to Reduce Intracellular pH in Vascular Smooth Muscle Cells

Wang

Chen

et al. 2019

Front. Pharmacol.

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Sulfur dioxide (SO 2 ) is a colorless and irritating gas. Recent studies indicate that SO 2 acts as the gas signal molecule and inhibits vascular smooth muscle cell (VSMC) proliferation. Cell proliferation depends on intracellular pH (pH i ). Transmembrane cystein mutation of Na + - independent Cl - /HCO 3 - exchanger (anion exchanger, AE) affects pH i . However, whether SO 2 inhibits VSMC proliferation by reducing pH i is still unknown. Here, we investigated whether SO 2 reduced pH i to inhibit the proliferation of VSMCs and explore its molecular mechanisms. Within a range of 50–200 μM, SO 2 was found to lower the pH i in VSMCs. Concurrently, NH 4 Cl pre-perfusion showed that SO 2 significantly activated AE, whereas the AE inhibitor 4,4′-diisothiocyanatostilbene- 2,20-disulfonic acid (DIDS) significantly attenuated the effect of SO 2 on pH i in VSMCs. While 200 μM SO 2 sulphenylated AE2, while dithiothreitol (DTT) blocked the sulphenylation of AE2 and subsequent AE activation by SO 2 , thereby restoring the pH i in VSMCs. Furthermore, DIDS pretreatment eliminated SO 2 -induced inhibition of PDGF-BB-stimulated VSMC proliferation. We report for the first time that SO 2 inhibits VSMC proliferation in part by direct activation of the AE via posttranslational sulphenylation and induction of intracellular acidification.

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“…Measurement of pH i was performed according to the literature (Lee et al, 2007; Galifianakis et al, 2018). Fluorescent indicator BCECF/AM was used to monitor pH i changes.…”

Section: Methodsmentioning

confidence: 99%

Sulfur Dioxide Activates Cl-/HCO3- Exchanger via Sulphenylating AE2 to Reduce Intracellular pH in Vascular Smooth Muscle Cells

Wang

Chen

et al. 2019

Front. Pharmacol.

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“…The most widely used chemical pH sensors are BCECF [43] and carboxy-SNARF-1 [44], as they have desirable photophysical properties for near-neutral intracellular H + concentration determination. Fluorescein and fluorescein derivatives such as carboxyfluorescein are common indicators of pH [45], but they quickly seep out of the cytosol through cell membranes and lead to inaccurate pH measurements.…”

Section: Chemical Biosensorsmentioning

confidence: 99%

The Cutting Edge of Disease Modeling: Synergy of Induced Pluripotent Stem Cell Technology and Genetically Encoded Biosensors

et al. 2021

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The development of cell models of human diseases based on induced pluripotent stem cells (iPSCs) and a cell therapy approach based on differentiated iPSC derivatives has provided a powerful stimulus in modern biomedical research development. Moreover, it led to the creation of personalized regenerative medicine. Due to this, in the last decade, the pathological mechanisms of many monogenic diseases at the cell level have been revealed, and clinical trials of various cell products derived from iPSCs have begun. However, it is necessary to reach a qualitatively new level of research with cell models of diseases based on iPSCs for more efficient searching and testing of drugs. Biosensor technology has a great application prospect together with iPSCs. Biosensors enable researchers to monitor ions, molecules, enzyme activities, and channel conformation in live cells and use them in live imaging and drug screening. These probes facilitate the measurement of steady-state concentrations or activity levels and the observation and quantification of in vivo flux and kinetics. Real-time monitoring of drug action in a specific cellular compartment, organ, or tissue type; the ability to screen at the single-cell resolution; and the elimination of the false-positive results caused by low drug bioavailability that is not detected by in vitro testing methods are a few of the benefits of using biosensors in drug screening. Here, we discuss the possibilities of using biosensor technology in combination with cell models based on human iPSCs and gene editing systems. Furthermore, we focus on the current achievements and problems of using these methods.

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Tailoring drug co-delivery nanosystem for mitigating U-87 stem cells drug resistance

Behrooz

Vazifehmand

Tajudin

et al. 2021

Drug Deliv. and Transl. Res.

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Intracellular pH Measurements in Glioblastoma Cells Using the pH-Sensitive Dye BCECF

Cited by 5 publications

References 6 publications

Sulfur Dioxide Activates Cl-/HCO3- Exchanger via Sulphenylating AE2 to Reduce Intracellular pH in Vascular Smooth Muscle Cells

Sulfur Dioxide Activates Cl-/HCO3- Exchanger via Sulphenylating AE2 to Reduce Intracellular pH in Vascular Smooth Muscle Cells

The Cutting Edge of Disease Modeling: Synergy of Induced Pluripotent Stem Cell Technology and Genetically Encoded Biosensors

Tailoring drug co-delivery nanosystem for mitigating U-87 stem cells drug resistance

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