To investigate in vivo the interconversion between glycine (Gly) and its N-methyl product sarcosine (Sar), [1-C-13]Gly and [1-C-14]Sar were infused into hourly fed pigs receiving diets with low- and high-threonine levels. An open two-pool model was developed to calculate Sar demethylation (DM) and Gly methylation (GM). During [1-C-14]Sar infusion, intracellular Gly specific radioactivities (SA) in the liver and kidney were higher than plasma Gly SA, suggesting that demethylation of Sar occurred in those tissues. DM estimated by using hippuric acid (HA) as the production pool had a mean value of 1.55-mu-mol.kg-1.h-1, similar to the Sar production rate (mean 1.85-mu-mol.kg-1.h-1). GM was undetectable (< 0.5-mu-mol.kg-1.h-1). These results suggest that, in fed pigs, Sar is produced mainly from choline catabolism and is degraded only to Gly in liver and kidney. On the assumption that Sar degradation gave rise only to Gly, the production rate of Gly (Gly PR) was calculated from [1-C-13]Gly and [1-C-14]Sar infusions using either the primary pools (plasma Gly and HA, respectively) or the secondary pools (HA and plasma Gly, respectively). The results were explained by a liver-plasma Gly exchange model. The whole body Gly irreversible loss, i.e., direct loss from plasma and liver, was calculated from this model to be 832 +/- 58-mu-mol.kg-1.h-1, showing that the estimation of Gly PR with [1-C-13]Gly infusion and plasma Gly enrichment (599 +/- 56-mu-mol.kg-1.h-1) was a significant underestimate of the true value. This model was also used to calculate a more realistic value of threonine disposal to Gly, showing that the use of an infusion of [1-C-13]Gly underestimated by 20% this component of threonine degradation.