Effectors of the frequency of calcium oscillations in HEK-293 cells: wavelet analysis and a computer model

Szekely, David, Brennan, Sarah C. ORCID: https://orcid.org/0000-0002-8719-4367, Mun, Hee-Chang, Conigrave, Arthur D. and Kuchel, Philip W. 2009. Effectors of the frequency of calcium oscillations in HEK-293 cells: wavelet analysis and a computer model. European Biophysics Journal 39 (1) , pp. 149-165. 10.1007/s00249-009-0469-2

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Abstract

Oscillations of the intracellular concentration of Ca2+ in cultured HEK-293 cells, which heterologously expressed the calcium-sensing receptor, were recorded with the fluorophore Fura-2 using fluorescence microscopy. HEK-293 cells are extremely sensitive to small perturbations in extracellular calcium concentrations. Resting cells were attached to cover slips and perifused with saline solution containing physiologically relevant extracellular Ca2+ concentrations in the range 0.5–5 mM. Acquired digitized images of the cells showed oscillatory fluctuations in the intracellular Ca2+ concentration over the time course, and were processed as a function of the change in Fura-2 excitation ratio and frequency at 12–37°C. Newly developed data processing techniques with wavelet analysis were used to estimate the frequency at which the rectified sinusoidal oscillations occurred; we estimated ~4 min−1 under normal conditions. Temperature variations revealed an Arrhenius relationship in oscillation frequency. A critical Ca2+ concentration of ~2 mM was estimated, below which oscillations did not occur. These data were used to develop a kinetic model of the system that was simulated using Mathematica; kinetic parameter values were adjusted to match the experimentally observed oscillations of intracellular Ca2+ concentration as a function of extracellular Ca2+ concentration, and temperature; and from these, limit cycles were obtained and control coefficients were estimated for all parameters.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Publisher:	Springer
ISSN:	0175-7571
Last Modified:	27 Oct 2022 09:26
URI:	https://orca.cardiff.ac.uk/id/eprint/65843

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