Effect of temperature on bradykinin-induced arachidonate release and calcium mobilization in vascular endothelial cells

Abstract

The effect of decreased temperature on Ca(2+)-dependent arachidonic acid release was studied in vascular endothelial cells by investigating bradykinin (BK)-stimulated Ca2+ mobilization, inositol phosphate formation and arachidonic acid release. At both 37 degrees C and 22 degrees C, BK efficiently increased cytosolic Ca2+ concn. ([Ca2+]i). At 22 degrees C, peak [Ca2+]i was higher and returned to basal levels more slowly. Although this response was preceded by rapid formation of Ins(1,4,5)P3, the activity of phospholipase C was significantly impaired at 22 degrees C. To determine if Ins(1,4,5)P3 effectively mobilized intracellular Ca2+, we used saponin-permeabilized cells. Ins(1,4,5)P3, mobilized sequestered Ca2+ to a similar degree at 37 degrees C and 22 degrees C, although Ca2+ release was prolonged at 22 degrees C. In intact cells, BK mobilized intracellular Ca2+ stores and activated Ca2+ entry. The rate of 45Ca2+ entry was approx. 2-fold slower at 22 degrees C, even though the peak and duration of the rise in [Ca2+]i were higher and sustained at the lower temperature. TG mobilized intracellular Ca2+, activated Ca2+ entry and elevated [Ca2+]i at both temperatures. As with BK, the peak [Ca2+]i reached after thapsigargin treatment was higher at 22 degrees C. This effect of lower temperature on [Ca2+]i was most probably due to decreased Ca2+ efflux after a decrease in activity of the Ca(2+)-ATPase on the plasma membrane. Both A23187 and BK were shown to stimulate phospholipase A2 and arachidonic acid release at 22 degrees C. In each case, the rate and extent of release were decreased compared with that at 37 degrees C. Among several effects, lowering the temperature decreases the activity of phospholipase C, Ca(2+)-ATPase(s), Ca(2+)-entry mechanisms and phospholipase A2. Together, these effects lead to a higher and more prolonged elevation of [Ca2+]i, but a decrease in arachidonate release in response to BK.