In this study we examined the effect of cholesterol (Diet 2), cholesterol and fish oil (FO) polyunsaturated acid (Diet 3), and polyunsaturated acid (Diet 4) enriched diets upon the acrosome reaction (AR) of New Zealand White rabbit spermatozoa. Male rabbits fed with cholesterol alone or with FO increased their cholesterol and LDL-cholesterol serum levels after 15 days of diet. Ten semen samples were obtained after 2 months of diet. Our results suggest that hypercholesterolemia and hypertriglyceridemia in male rabbits could produce a decreased capacity of sperm AR after 4 h (0%, 0%, and 60% lower than the control), 6 h (0%, 68%, and 44%), or 8 h (58%, 52% and 32%) of incubation in capacitating medium. Another set of experiments were made with lysophosphatidylcholine (LPC), 80 micrograms/ml, and the same pattern of AR was seen. Nevertheless, the high cholesterol and total lipids (TL) levels in serum did not affect the cholesterol levels in seminal plasma (SP) but affect the SP total lipids. The diminished capacity of rabbit sperm to undergo the AR was not reverted by in vitro incubation with the Shinitsky medium for cholesterol depletion (MDC). These results indirectly suggest that the cholesterol/phospholipid ratio in hypercholesterolemic sperm is similar to that of controls and are in agreement with preliminary studies made in our laboratory that evidenced the same cholesterol/phospholipid ratio in rabbit sperm from hypercholesterolemic animals than from controls.(ABSTRACT TRUNCATED AT 250 WORDS)