Evidence for the existence of inositol (1,4,5)-trisphosphate- and ryanodine-sensitive pools in bovine endothelial cells. Ca2+ releases in cells with different basal level of intracellular Ca2+.

Abstract

In single bovine aortic endothelial (BAE) cells pre-loaded with Fura-2, Ca2+ transients in a Ca2+-free medium have been revealed, which evidently reflects Ca2+ release from intracellular stores. In cells with different levels of resting basal cytoplasmic Ca2+ ([Ca2+]i) from about 50 to 110 nM, a biphasic dependence of the Ca2+ transients on resting [Ca2+]i was shown and spontaneous Ca2+ oscillations were observed. At a [Ca2+]i level over 110 nM, a pronounced rise in Ca2+ transients occurred and only single transients were observed. Ryanodine (10 microM) produced a transient [Ca2+]i elevation, suggesting the presence of ryanodine receptors in intracellular store membranes. The results imply that both inositol 1,4,5-trisphosphate-sensitive Ca2+ release (IICR) and Ca2+-sensitive Ca2+ release (CICR) take place in BAE cells. Only IICR seems to be sufficient for generating baseline Ca2+ oscillations in BAE cells, whereas the ATP-induced (5-100 microM) Ca2+ response involves the CICR set in motion by an oscillatory IICR of high frequency. The completion of both the spontaneous and ATP-induced Ca2+ transients was associated with a [Ca2+]i decrease to a level below the initial resting [Ca2+]i (undershoot). Its depth biphasically depended on the resting [Ca2+]i from 50 to 110 nM, suggesting that the lack of a Ca2+ leak from inositol 1,4,5-trisphosphate-sensitive stores is responsible for the undershoot in this range. The Ca2+ leak is concluded to play a key role in the initiation and termination of regenerative IICR both in spontaneous oscillations and in ATP-induced transients.