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Biophys. J. BioFAST: First Published January 28, 2005. doi:10.1529/biophysj.104.045260
© 2005 by the Biophysical Society.

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BIOPHYSICAL THEORY AND MODELING

A model-independent algorithm to derive Ca2+ fluxes underlying local cytosolic Ca2+ transients

Alejandra C. Ventura 1, Luciana Bruno 1, Angelo Demuro 2, Ian Parker 2 and Silvina Ponce Dawson 1*

1 Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires
2 University of California, Irvine

* To whom correspondence should be addressed. E-mail: silvina{at}df.uba.ar.

Submitted on May 6, 2004
Revised on June 14, 2004
Accepted on 12 January 2005


  Abstract
Local intracellular Ca2+ signals result from Ca2+ flux into the cytosol through individual channels or clusters of channels. To gain a mechanistic understanding of these events we need to know the magnitude and spatial distribution of the underlying Ca2+ flux. However, this is difficult to infer from fluorescence Ca2+ images because the distribution of Ca2+-bound dye is affected by poorly characterized processes including diffusion of Ca2+ ions, their binding to mobile and immobile buffers and sequestration by Ca2+ pumps. Several methods have previously been proposed to derive Ca2+ flux from fluorescence images, but all require explicit knowledge or assumptions regarding these processes. We now present a novel algorithm that requires few assumptions and is largely model-independent. By testing the algorithm with both numerically generated ``image'' data and experimental images of sparklets resulting from Ca2+ flux through individual voltage-gated channels, we show that it satisfactorily reconstructs the magnitude and time course of the underlying Ca2+ currents.

Key Words: buffers, calcium fluxes, calcium transients, computer algorithm, diffusion, fluorescent images




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Copyright © 2005 by the Biophysical Society.