Brewer's yeast was grown on a defined medium containing glucose, ammonia, salts and vitamins plus tracer 51Cr without (low-Cr) or with (high-Cr) carrier Cr. The two batches of yeast differed by more than 100-fold in Cr content, containing 80 ng and 10 7mu;g Cr/g dry yeast respectively. Extraction and fractionation procedures were designed to isolate Cr complexes with properties similar to those reported for glucose tolerance factor. After weaning, rats were reared on rat cubes (normal diet) or on a diet containing less than 0.1 μg Cr/kg (low-Cr diet), or on the low-Cr diet supplemented with Cr (1 mg Cr/kg). Hepatocytes from these rats were incubated with [U-14C]glucose and incorporation of 14C into glycogen was measured. Incorporation of glucose-C into glycogen was enhanced by some yeast fractions in the presence of insulin, but had less effect in the absence of insulin. No difference could be detected between the responses to fractions from high- or low-Cr yeast extracts, or between responses by hepatocytes from animals fed on normal or low-Cr diets with or without Cr upplementation. Glycogen synthetase (EC 2.4.1.11) activity (total and percentage in the a form) was similar in hepatocytes isolated from animals on the normal and low-Cr diets. Those yeast fractions which enhanced the response to insulin in the 14C-incorporation experiments also enhanced the percentage of the enzyme in the a form in the presence of insulin, but not in the absence of insulin. The presence in yeast extracts of material which enhances the response to insulin by hepatocytes may help to explain the reported beneficial effects of dietary yeast supplements on glucose tolerance.