Stable isotope studies in conscious mice have been limited by the invasive catheterization procedures and relatively large sample size required. We developed minimally invasive catheterization protocols that together with the ability to analyze small samples have allowed for the study of urea kinetics in conscious mice. A single dose of 15N15N-urea followed by multiple sampling in mice (n = 6) showed that a primary pool of urea exchanged rapidly [70.65 +/- 14.96 mmol/(kg x h)] with a secondary pool. The urea entry rate determined with this protocol was 3.36 +/- 0.30 mmol/(kg x h). Continuous infusion of 15N15N-urea (n = 6) achieved plateau enrichment values at 3.3 +/- 0.2.h from which the urea entry rate was determined by isotope dilution [3.24 +/- 0.23 mmol/(kg x h)]. The urea entry rate measured by the single dose or continuous infusion protocol did not differ (P = 0.76). The minimally invasive methods described allow us to study not only ureagenesis and urea cycle disorders in vivo, but also urea transport and transporter function and nitrogen metabolism in general in mouse models. This is especially relevant because mouse targeting technologies will likely facilitate the generation of organ and tissue specific nulls of the various urea cycle enzymes.