Arachidonic acid (AA) release and prostaglandin (PG) biosynthesis were studied in rat embryo fibroblasts (R 129) synchronized by double thymidine-excess block. Whatever the culture medium was (medium 199 supplemented with 10% fetal calf serum (FCS) or 1% FCS plus 0.1% bovine serum albumin (BSA], AA release rapidly increased until the 4th hour of the cell cycle (S phase), remained on a plateau in G2M and G1 phases and did not increase again in the S phase of the following cell cycle. Time course and amplitude of AA release in synchronized cells did not differ from what it was observed after the simple renewal of the culture medium in asynchronous cells. So AA release seemed to be independent of the cell cycle. By contrast, PGE2 and PGF2 alpha biosynthesis clearly increased in the S phase of two consecutive cell cycles, indicating that cyclooxygenase activity and not phospholipase A2 activity vary according to the cell cycle.