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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
Authors:Camilla Rozanski  Michelle Belton  Frank S. Prato  Jeffrey J.L. Carson
Affiliation:1. Imaging Program, Lawson Health Research Institute, London, Ontario, Canada;2. Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada
Abstract: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 ([Ca2+]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. [Ca2+]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 [Ca2+]c time series included: average [Ca2+]c during the Pre‐Field and Field Conditions, peak [Ca2+]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 [Ca2+]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 [Ca2+]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. Bioelectromagnetics 30:213–221, 2009. © 2008 Wiley‐Liss, Inc.
Keywords:DC  fluorescence  ratiometric  fura‐2  free radical
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