Fertilization of rat eggs in vitro could not be achieved when epididymal spermatozoa were preincubated and eggs in clots incubated in a chemically defined medium without D-glucose. Very high proportions (84-100%) of eggs examined were undergoing fertilization when 2.78-8.34 mM-D-glucose were included in the medium. The substitution of D-fructose or D-galactose for D-glucose resulted in very poor penetration rates (0-4%), but D-mannose was effective for fertilization (59-99% penetration). Incubations for sperm capacitation and egg fertilization in different media containing the various hexoses showed that rat epididymal spermatozoa could be partly capacitated without hexose, that D-glucose, D-mannose or D-galactose but not D-fructose is effective for sperm capacitation and that only D-glucose and D-mannose supported penetration of eggs in vitro.