Real‐time measurement of cytosolic free calcium concentration in HL‐60 cells during static magnetic field exposure and activation by ATP

Belton, Michelle; Commerford, Kristy; Hall, Justin; Prato, Frank S.; Carson, Jeffrey J. L.

doi:10.1002/bem.20409

Cited by 20 publications

(20 citation statements)

References 32 publications

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“…In contrast, some studies have found no effect of SMF exposure on calcium homeostasis. For example, in a previous study, we examined whether a 100 mT SMF would affect the [Ca 2þ ] c response to ATP [Belton et al, 2008]. Upon addition of ATP to fura 2-AM loaded HL-60 cells we observed a large and sudden increase in [Ca 2þ ] c .…”

Section: Introductionmentioning

confidence: 86%

“…Cytosolic free calcium in fura-2 loaded cell samples was measured with a modified ratiometric fluorescence spectroscopy system (PTI, London, ON) comprised of a light source, chopper with filter assembly, bifurcated quartz fiber bundle, control electronics, a photomultiplier tube detector, and a personal computer [Belton et al, 2008]. The data collection and spectrometer operation were handled by commercial software (Felix32, PTI, Lawrenceville, NJ).…”

Section: Apparatus For Measurement Of Cytosolic Free Calciummentioning

confidence: 99%

“…Bell model 9500, Sypris Test & Measurement, Orlando, FL) and found to be within AE5% of the mean flux density. More details on the design of the system can be found in Belton et al [2008]. Cells were maintained in suspension with a computer-controlled mixer similar in design to previously described systems [Grundler et al, 1977].…”

Section: Apparatus For Measurement Of Cytosolic Free Calciummentioning

confidence: 99%

“…Other metrics were considered (e.g., slope of the ATP response), but the metrics presented capture the overall features of each time series and are directly comparable to our previous observations [Belton et al, 2008]. All time series calculations were performed with custom software written in Cþþ (Microsoft Visual Cþþ).…”

Section: Time Series and Statistical Analysismentioning

confidence: 99%

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Effect of 100 mT homogeneous static magnetic field on [Ca²⁺]_c response to ATP in HL‐60 cells following GSH depletion

Belton

Prato

Rozanski

et al. 2009

Bioelectromagnetics

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Calcium is an important molecule in a number of biological systems. Often these systems are signal transduction cascades involving molecules such as ATP. ATP activates second messengers which can interact with ion channels on the endoplasmic/sarcoplasmic reticulum resulting in the emptying of the intracellular calcium stores and an increase in cytosolic free calcium concentration ([Ca2+]c). Changes in [Ca2+]c can be influenced by external factors such as a static magnetic field (SMF). One hypothesis suggests that a SMF affects the cells through the radical pair mechanism. By reducing the number of antioxidant molecules like glutathione (GSH), the proportion of free radicals in the cells is increased and may lead to a greater probability of a biological response to a SMF. The purpose of this study was to determine if the [Ca2+]c response to ATP was affected by depletion of GSH by diethylmaleate (DEM) and the absence or presence of a 100 mT homogeneous SMF. Undifferentiated HL-60 cells were loaded with fura-2 AM. [Ca2+]c was measured in real time using a ratiometric fluorescence spectroscopy system. Various (DEM) ranging from 1 to 15 mM were added to deplete GSH. Cells were either exposed to sham or magnetic field (100 mT) for 13 min (780 s) and challenged with 1 microM ATP. The data show that [Ca2+]c was elevated following treatment with DEM with greater [Ca2+]c at higher [DEM]. The [Ca2+]c response to ATP was decreased as the DEM concentration increased. However, there was no effect of a 100 mT SMF on the average [Ca2+]c peak following ATP activation or the full width at half maximum (FWHM) of the [Ca2+]c response and recovery after ATP activation.

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Section: Introductionmentioning

confidence: 86%

Section: Apparatus For Measurement Of Cytosolic Free Calciummentioning

confidence: 99%

Section: Apparatus For Measurement Of Cytosolic Free Calciummentioning

confidence: 99%

Section: Time Series and Statistical Analysismentioning

confidence: 99%

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Effect of 100 mT homogeneous static magnetic field on [Ca²⁺]_c response to ATP in HL‐60 cells following GSH depletion

Belton

Prato

Rozanski

et al. 2009

Bioelectromagnetics

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“…ATP induced apoptotic cell death of both HL-60 and F-36P leukaemia cell lines [456]. Static magnetic field exposure of HL-60 cells increased the increase in [Ca 2+ ] i produced by ATP [457,458]. A 3 receptor agonists were shown to inhibit proliferation and induce apoptosis of HL-60 leukaemia cells [459,460].…”

Section: Lymphocytic Leukaemiamentioning

confidence: 98%

Purinergic signalling and cancer

Burnstock

Virgilio

2013

Purinergic Signalling

266

265

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Receptors for extracellular nucleotides are widely expressed by mammalian cells. They mediate a large array of responses ranging from growth stimulation to apoptosis, from chemotaxis to cell differentiation and from nociception to cytokine release, as well as neurotransmission. Pharma industry is involved in the development and clinical testing of drugs selectively targeting the different P1 nucleoside and P2 nucleotide receptor subtypes. As described in detail in the present review, P2 receptors are expressed by all tumours, in some cases to a very high level. Activation or inhibition of selected P2 receptor subtypes brings about cancer cell death or growth inhibition. The field has been largely neglected by current research in oncology, yet the evidence presented in this review, most of which is based on in vitro studies, although with a limited amount from in vivo experiments and human studies, warrants further efforts to explore the therapeutic potential of purinoceptor targeting in cancer.

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Real‐time measurement of cytosolic free calcium concentration in DEM‐treated HL‐60 cells during static magnetic field exposure and activation by ATP

Rozanski¹,

Belton²,

Prato³

et al. 2008

Bioelectromagnetics

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This study investigated whether glutathione depletion affected the sensitivity of HL-60 cells to static magnetic fields. The effect of Diethylmaleate (DEM) on static magnetic field induced changes in cytosolic free calcium concentration ([Ca(2+)](c)) was examined. Cells were loaded with a fluorescent dye and exposed to a uniform static magnetic field at a strength of 0 mT (sham) or 100 mT. [Ca(2+)](c) was monitored during field and sham exposure using a ratiometric fluorescence spectroscopy system. Cells were activated by the addition of ATP. Metrics extracted from the [Ca(2+)](c) time series included: average [Ca(2+)](c) during the Pre-Field and Field Conditions, peak [Ca(2+)](c) following ATP activation and the full width at half maximum (FWHM) of the peak ATP response. Comparison of each calcium metric between the sham and 100 mT experiments revealed the following results: average [Ca(2+)](c) measured during the Field condition was 53 +/- 2 nM and 58 +/- 2 nM for sham and 100 mT groups, respectively. Average FWHM was 51 +/- 3 s and 54 +/- 3 s for sham and 100 mT groups, respectively. An effect of experimental order on the peak [Ca(2+)](c) response to ATP in sham/sham experiments complicated the statistical analysis and did not allow pooling of the first and second order experiments. No statistically significant difference between the sham and 100 mT groups was observed for any of the calcium metrics. These data suggested that manipulation of free radical buffering capacity in HL-60 cells did not affect the sensitivity of the cells to a 100 mT static magnetic field.

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Real‐time measurement of cytosolic free calcium concentration in HL‐60 cells during static magnetic field exposure and activation by ATP

Cited by 20 publications

References 32 publications

Effect of 100 mT homogeneous static magnetic field on [Ca²⁺]_c response to ATP in HL‐60 cells following GSH depletion

Effect of 100 mT homogeneous static magnetic field on [Ca²⁺]_c response to ATP in HL‐60 cells following GSH depletion

Purinergic signalling and cancer

Real‐time measurement of cytosolic free calcium concentration in DEM‐treated HL‐60 cells during static magnetic field exposure and activation by ATP

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Real‐time measurement of cytosolic free calcium concentration in HL‐60 cells during static magnetic field exposure and activation by ATP

Cited by 20 publications

References 32 publications

Effect of 100 mT homogeneous static magnetic field on [Ca2+]c response to ATP in HL‐60 cells following GSH depletion

Effect of 100 mT homogeneous static magnetic field on [Ca2+]c response to ATP in HL‐60 cells following GSH depletion

Purinergic signalling and cancer

Real‐time measurement of cytosolic free calcium concentration in DEM‐treated HL‐60 cells during static magnetic field exposure and activation by ATP

Contact Info

Product

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About

Effect of 100 mT homogeneous static magnetic field on [Ca²⁺]_c response to ATP in HL‐60 cells following GSH depletion

Effect of 100 mT homogeneous static magnetic field on [Ca²⁺]_c response to ATP in HL‐60 cells following GSH depletion