SPONTANEOUS CA2+ RELEASE FROM A CAFFEINE AND RYANODINE-SENSITIVE INTRACELLULAR CA2+ STORE IN FRESHLY PREPARED HEPATOCYTES

A considerable fraction of freshly prepared hepatocytes loaded with the fluorescent [Ca2+](i) indicator fura-a exhibited spontaneous rhythmic fluctuations that tended to decrease with increasing length of incubation after isolation. These oscillations were dependent on the external Ca2+. They could no longer be observed when a Ca2+ chelator- (ethylenebis [oxyethylenenitrilo]) tetraacetic acid-was added to medium. Addition of thapsigargin, which is known to release Ca2+ from inositol 1,4,5-trisphosphate-sensitive and -insensitive Ca2+ stores, induced a large transient increase in [Ca-2+](i) and abolished the fluctuations. When the cells were treated with 2 mmol/L caffeine, frequency was increased, whereas 10 mmol/L caffeine induced a single large peak followed by a persistent plateau. Moreover, addition of dibutyryl cAMP led to decreased frequency of fluctuations. Ryanodine caused larger fluctuations; thereafter the [Ca2+](i) level became much higher and the spices ceased. These results suggest that spontaneous rhythmic fluctuations in freshly prepared hepatocytes are driven by Ca2+ release from a caffeine- and ryanodine-sensitive calcium-induced calcium release pool.