Cauda epididymal spermatozoa were examined for their capacity to synthesize prostaglandins (PGs) and hydroxy acids and the role of nitric oxide (NO) on cyclooxygenase and lipoxygenase enzyme activity was determined. Sperm suspensions were incubated in the presence or absence of 300 μM sodium nitroprusside (NP) and 20 μg/ml hemoglobin (Hb) and then incubated with [ 1 4 C]arachidonic acid (AA) for 60 min. The cyclooxygenase and lipoxygenase products of AA metabolism were measured for their radioactivity. It was found that mouse spermatozoa could synthesize PGE 2 and 5-hydroxyeicosatetraenoic acid (5-HETE) under these experimental conditions. The synthesis of these two metabolites was 100% stimulated by NP, the releaser of NO, and the response to NP was completely prevented by Hb, a scavenger of NO. These data provide evidence that mouse spermatozoa can synthesize PGE 2 and 5-HETE in the presence of AA in vitro and that NO is involved in the production of AA metabolites in the male gamete.