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In recent years, several metabolic roles have been proposed for vitamin C. Recent information suggests a strong causal relationship between high endogenous levels of ascorbic acid and changes in normal reproductive biology. Using highly enriched populations of guinea pig Leydig cells, we have found that elevated levels 50 to 500 microM of ascorbate significantly P < 0.01 depressed testosterone production in a dose-dependent manner while low levels 0 to 10 microM were without effect. Leydig cells incubated under hypoxic 3% oxygen culture conditions produced significantly P < 0.01 more testosterone than similar cells cultured under normoxic 19% oxygen conditions. The results of this study suggest that high concentrations of ascorbate and normoxic culture conditions suppress testosterone production in isolated Leydig cells. Thus, it would seem that there exists a delicate balance between normal metabolic requirements for vitamin C and excessive ascorbate levels that might alter normal gonadal reproductive events.