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Biophys J, July 2000, p. 163-183, Vol. 79, No. 1

An Image-Based Model of Calcium Waves in Differentiated Neuroblastoma Cells

Charles C. Fink,* Boris Slepchenko,dagger Ion I. Moraru,Dagger James Watras,§ James C. Schaff,dagger and Leslie M. Loew*dagger

 *Department of Physiology,  dagger Center for Biomedical Imaging Technology,  Dagger Department of Surgery, and  §Department of Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030 USA

Calcium waves produced by bradykinin-induced inositol-1,4,5-trisphosphate (InsP3)-mediated release from endoplasmic reticulum (ER) have been imaged in N1E-115 neuroblastoma cells. A model of this process was built using the "virtual cell," a general computational system for integrating experimental image, biochemical, and electrophysiological data. The model geometry was based on a cell for which the calcium wave had been experimentally recorded. The distributions of the relevant cellular components [InsP3 receptor (InsP3R)], sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA) pumps, bradykinin receptors, and ER] were based on 3D confocal immunofluorescence images. Wherever possible, known biochemical and electrophysiological data were used to constrain the model. The simulation closely matched the spatial and temporal characteristics of the experimental calcium wave. Predictions on different patterns of calcium signals after InsP3 uncaging or for different cell geometries were confirmed experimentally, thus helping to validate the model. Models in which the spatial distributions of key components are altered suggest that initiation of the wave in the center of the neurite derives from an interplay of soma-biased ER distribution and InsP3 generation biased toward the neurite. Simulations demonstrate that mobile buffers (like the indicator fura-2) significantly delay initiation and lower the amplitude of the wave. Analysis of the role played by calcium diffusion indicated that the speed of the wave is only slightly dependent on the ability of calcium to diffuse to and activate neighboring InsP3 receptor sites.

Biophys J, July 2000, p. 163-183, Vol. 79, No. 1
© 2000 by the Biophysical Society   0006-3495/00/07/163/21  $2.00


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