INOSITOL 1,4,5-TRISPHOSPHATE-SENSITIVE AND CAFFEINE-SENSITIVE CA2-STORING ORGANELLE IN BOVINE ADRENAL CHROMAFFIN CELLS()

The uptake and release properties of Ca2+ by several subcellular fract ions of the bovine adrenal medulla were investigated. Investigation by the Ca-45(2+) tracer method showed that permeabilized cells and the f ractions of mitochondria (MT) and microsomes (MC) caused ATP-dependent Ca2+ uptake in a Ca2+ concentration-dependent manner (pCa 8 - 4), whe reas permeabilized cells and the fractions of secretory granules (SG) were able to accumulate a significant amount of Ca2+ even in the absen ce of ATP, which was completed by the addition of hexokinase and gluco se. In these organelle fractions, Ca2+ uptake in the presence of ATP a t pCa 7 and pCa 5.8 was well-correlated with the activity of the NADPH cytochrome c reductase (marker enzyme for the endoplasmic reticulum) and cytochrome c oxidase (marker enzyme for mitochondria), respectivel y. As detected by Fura-2 ratiometry, both inositol 1,4,5-trisphosphate (IP3) and caffeine caused concentration-dependent Ca2+ releases from permeabilized cells and MC, but not from MT and SG. In an ATP-depleted condition, homogenates still took up a significant amount of Ca2+ but was not able to respond to IP3 and caffeine. These results suggest th at the endoplasmic reticulum is a major Ca2+-storing organelle, which releases Ca2+ in response to IP3 and caffeine in bovine adrenal chroma ffin cells.