Differential regulation of ER Ca2+ uptake and release rates accounts for multiple modes of Ca2+-induced Ca2+ release

The ER is a central element in Ca(2+) signaling, both as a modulator of cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) and as a locus of Ca(2+)-regulated events. During surface membrane depolarization in excitable cells, the ER may either accumulate or release net Ca(2+), but the conditions of stimulation that determine which form of net Ca(2+) transport occurs are not well understood. The direction of net ER Ca(2+) transport depends on the relative rates of Ca(2+) uptake and release via distinct pathways that are differentially regulated by Ca(2+), so we investigated these rates and their sensitivity to Ca(2+) using sympathetic neurons as model cells. The rate of Ca(2+) uptake by SERCAs (J(SERCA)), measured as the t-BuBHQ-sensitive component of the total cytoplasmic Ca(2+) flux, increased monotonically with [Ca(2+)](i). Measurement of the rate of Ca(2+) release (J(Release)) during t-BuBHQ-induced [Ca(2+)](i) transients made it possible to characterize the Ca(2+) permeability of the ER ((~)P(ER)), describing the activity of all Ca(2+)-permeable channels that contribute to passive ER Ca(2+) release, including ryanodine-sensitive Ca(2+) release channels (RyRs) that are responsible for CICR. Simulations based on experimentally determined descriptions of J(SERCA), and of Ca(2+) extrusion across the plasma membrane (J(pm)) accounted for our previous finding that during weak depolarization, the ER accumulates Ca(2+), but at a rate that is attenuated by activation of a CICR pathway operating in parallel with SERCAs to regulate net ER Ca(2+) transport. Caffeine greatly increased the [Ca(2+)] sensitivity of ((~)P(ER)), accounting for the effects of caffeine on depolarization-evoked [Ca(2+)](i) elevations and caffeine-induced [Ca(2+)](i) oscillations. Extending the rate descriptions of J(SERCA), ((~)P(ER)), and J(pm) to higher [Ca(2+)](i) levels shows how the interplay between Ca(2+) transport systems with different Ca(2+) sensitivities accounts for the different modes of CICR over different ranges of [Ca(2+)](i) during stimulation.