Correlation among endothelial cell shape, F-actin arrangement, and prostacyclin synthesis.

Though many factors have been identified which modulate prostacyclin (PGI2) synthesis, there is little information on cellular mechanisms whereby endothelial cells (EC) regulate their basal eicosanoid metabolism. Using substrates of various adhesive capacities, bovine and porcine aortic EC shape and cytoskeletal F-actin arrangement could be modulated. Staining with rhodamine-phalloidin (R-P) permitted analysis of F-actin arrangement, while differences in cell shape were determined by measurement of cell perimeter surface area (CPSA). Spectrophotoflurometric measurements were used to quantitate the R-P binding capacity of the cultures. Cultures of reduced CPSA (225.2 +/- 13.5 mu2) generated the highest levels of basal PGl2 (6.14 +/- 0.51 pg/ug cell protein); had a diffuse arrangement of F-actin and an increased binding capacity for R-P (463.55 +/- 50.58 nmoles/ug cell protein). Cultures of enlarged CPSA (1399.3 +/- 148.3 mu2), with many actin cables and a significantly reduced (p less than 0.001) R-P binding capacity (74.941 +/- 11.79 nmoles/ug of cell protein) produced significantly smaller (p less than 0.001) basal quantities of PGl2 (1.33 +/- 0.14 pg/ug cell protein). Similarly, arachidonic acid stimulation of cultures of reduced CPSA resulted in an increased synthesis of PGl2 when compared to stimulated cultures of enlarged cells. These findings suggest a role for cell shape and the cytoskeleton in the mechanism controlling PGl2 production and indicate that alteration of the arrangement of F-actin may be of importance in regulation of EC eicosanoid metabolism.