The metabolism of lactate, alanine and glutamine was studied in isolated canine liver cells. Lactate proved a better gluconeogenic precursor than alanine and glutamine. Glucose was synthesized at similar rates from both amino acids and ureogenesis from glutamine was double that from alanine. As in isolated rat liver cells, glucose and urea represented the main carbon and nitrogenous end-products of lactate, alanine and glutamine metabolism. Taken with the known levels of hepatic uptake of glutamine and alanine, these findings suggest that in the fasted dog, unlike in the fasted rat, glutamine and not alanine is the main transporter of nitrogen and carbon from muscle to liver for urea synthesis and gluconeogenesis.