Using two dyes to observe the competition of Ca2+ trapping mechanisms and their effect on intracellular Ca2+ signals.

The specificity and universality of intracellular [Formula: see text] signals rely on the variety of spatio-temporal patterns that the [Formula: see text] concentration can display. [Formula: see text] liberation through inositol 1,4,5-trisphosphate receptors ([Formula: see text]) is key for this variety. In this paper, we study how the competition between buffers of different kinetics affects [Formula: see text] signals that involve [Formula: see text] release through [Formula: see text]. The study also provides insight into the underlying spatial distribution of the channels that participate in the signals. Previous works on the effects of [Formula: see text] buffers have drawn conclusions 'indirectly' by observing the [Formula: see text]-bound dye distributions in the presence of varying concentrations of exogenous buffers and using simulations to interpret the results. In this paper, we make visible the invisible by observing the signals simultaneously with two dyes, [Formula: see text] and [Formula: see text], each of which plays the role of a slow or fast [Formula: see text] buffer, respectively. Our observations obtained for different concentrations of [Formula: see text] highlight the dual role that fast buffers exert on the dynamics, either reducing the intracluster channel coupling or preventing channel inhibition and allowing the occurrence of relatively long cycles of [Formula: see text] release. Our experiments also show that signals with relatively high [Formula: see text] release rates remain localized in the presence of large [Formula: see text] concentrations, while the mean speed of the elicited waves increases. We interpret this as a consequence of the more effective uncoupling between [Formula: see text] clusters as the slow dye concentration increases. Combining the analysis of the experiments with numerical simulations, we also conclude that [Formula: see text] release not only occurs within the close vicinity of the centers of the clearly identifiable release sites ([Formula: see text] clusters) but there are also functional [Formula: see text] in between them.

Simplified model of cytosolic Ca2+ dynamics in the presence of one or several clusters of Ca2+ -release channels.

Calcium release from intracellular stores plays a key role in the regulation of a variety of cellular activities. In various cell types this release occurs through inositol-triphosphate (IP3) receptors which are Ca2+ channels whose open probability is modulated by the cytosolic Ca2+ concentration itself. Thus, the combination of Ca2+ release and Ca2+ diffusion evokes a variety of Ca2+ signals depending on the number and relative location of the channels that participate of them. In fact, a hierarchy of Ca2+ signals has been observed in Xenopus laevis oocytes, ranging from very localized events (puffs and blips) to waves that propagate throughout the cell. In this cell type channels are organized in clusters. The behavior of individual channels within a cluster cannot be resolved with current optical techniques. Therefore, a combination of experiments and mathematical modeling is unavoidable to understand these signals. However, the numerical simulation of a detailed mathematical model of the problem is very hard given the large range of spatial and temporal scales that must be covered. In this paper we present an alternative model in which the cluster region is modeled using a relatively fine grid but where several approximations are made to compute the cytosolic Ca2+ concentration ([Ca;{2+}]) distribution. The inner-cluster [Ca;{2+}] distribution is used to determine the openings and closings of the channels of the cluster. The spatiotemporal [Ca;{2+}] distribution outside the cluster is determined using a coarser grid in which each (active) cluster is represented by a point source whose current is proportional to the number of open channels determined before. A full reaction-diffusion system is solved on this coarser grid.

Computing with excitable systems in a noisy environment.

In this work we show that excitable units with biologically inspired couplings are capable of performing any logic operation in a noisy environment without synchronization.

Saltatory propagation of Ca2+ waves by Ca2+ sparks.

Punctate releases of Ca2+, called Ca2+ sparks, originate at the regular array of t-tubules in cardiac myocytes and skeletal muscle. During Ca2+ overload sparks serve as sites for the initiation and propagation of Ca2+ waves in myocytes. Computer simulations of spark-mediated waves are performed with model release sites that reproduce the adaptive Ca2+ release observed for the ryanodine receptor. The speed of these waves is proportional to the diffusion constant of Ca2+, D, rather than D, as is true for reaction-diffusion equations in a continuous excitable medium. A simplified "fire-diffuse-fire" model that mimics the properties of Ca2+-induced Ca2+ release (CICR) from isolated sites is used to explain this saltatory mode of wave propagation. Saltatory and continuous wave propagation can be differentiated by the temperature and Ca2+ buffer dependence of wave speed.