Ca uptake by heart cells: I. Ca uptake by the sarcoplasmic reticulum of intact heart cells in suspension

Abstract

Electric field stimulation of adult rat heart cells suspended in medium with 0.2 mM Ca and isoproterenol caused 45Ca uptake at a rate (5.25 pmol/mg/beat) proportional to stimulation frequency. Uptake was strongly inhibited by verapamil or thapsigargin. 45Ca autoradiography showed that stimulation dependent verapamil sensitive uptake was associated with the rod shaped cells, while the uptake by round cells was unaffected by stimulation and was verapamil-insensitive. 45Ca efflux measurements revealed a caffeine-sensitive component of uptake which was abolished by thapsigargin, and a caffeine-insensitive component. Part of the latter was sensitive to thapsigargin but not to 30 s of stimulation; another part was sensitive to such stimulation but not to thapsigargin. With longer times of stimulation, the caffeine-insensitive pool increased in size, part of which appeared to be mitochondrial Ca uptake via a thapsigargin-sensitive pool. The caffeine-sensitive pool labelled quickly in stimulated cells and its size and rate of labelling was increased by stimulation frequency (3.87 pmol/mg/beat), while the caffeine-insensitive pool labelled more slowly and was relatively insensitive to stimulation (0.77 pmol/mg/beat). We conclude that essentially all of the SR Ca pool, as defined by its involvement in excitation-contraction coupling, is released by caffeine.