Quantitative analysis of mitochondrial Ca2+ uptake and release pathways insympathetic neurons - Reconstruction of the recovery after depolarization-evoked [Ca2+](i) elevations

Rate equations for mitochondrial Ca2+ uptake and release and plasma membran e Ca2+ transport were determined from the measured fluxes in the preceding study and incorporated into a model of Ca2+ dynamics. It was asked if the m easured fluxes are sufficient to account for the [Ca2+](i), recovery kineti cs after depolarization-evoked [Ca2+](i) elevations. Ca2+ transport across the plasma membrane was described by a parallel extrusion/leak system, whil e the rates of mitochondrial Ca2+ uptake and release were represented using equations like those describing Ca2+ transport by isolated mitochondria. T aken together, these rate descriptions account very well for the time cours e of recovery after [Ca2+](i) elevations evoked by weak and strong depolari zation and their differential sensitivity to FCCP, CGP 37157, and [Na+](i). The model also leads to three general conclusions about mitochondrial Ca2 transport in intact cells: (1) mitochondria are expected to accumulate Ca2 + even in response to stimuli that raise [Ca2+](i) only slightly above rest ing levels; (2) there are two qualitatively different stimulus regimes that parallel the buffering and non-buffering modes of Ca2+ transport by isolat ed mitochondria that have been described previously; (3) the impact of mito chondrial Ca2+ transport on intracellular calcium dynamics is strongly infl uenced by nonmitochondrial Ca2+ transport; in particular, the magnitude of the prolonged [Ca2+](i) elevation that occurs during the plateau phase of r ecovery is related to the Ca2+ set-point described in studies of isolated m itochondria, but is a property of mitochondrial Ca2+ transport in a cellula r context. Finally, the model resolves the paradoxical finding that stimulu s-induced [Ca2+](i) elevations as small as similar to 300 nM increase intra mitochondrial total Ca2+ concentration, but die steady [Ca2+](i) elevations evoked by such stimuli are not influenced by FCCP.