Comparison of constant infusion and flooding dose techniques to measure muscle protein synthesis rate in dogs

Abstract

The exact relationship between the constant infusion and flooding dose techniques for measuring fractional muscle protein synthesis (FMPS) rates was determined in anesthetized dogs during the postabsorptive period, using different isotopes of leucine as tracers. First, [1,2-13C]- and [U- 14C]leucine were given simultaneously by prime-constant infusion for 5 h to establish that both isotopes gave comparable FMPS rate values. The tracers gave almost identical FMPS rates and the values obtained after 3 and 5 h were not different. Next, the constant infusion and flooding dose methods were compared in the same animal by first measuring FMPS rate with a prime- constant infusion of [U-14C]leucine, then with a flooding dose of 376 μmol/kg [1,2-13C]leucine. The flooding dose method gave FMPS rates that were markedly greater than the constant infusion values. Finally, the effect of a 376 μmol/kg leucine flooding dose on FMPS rate was assessed using the prime-constant infusion of [U-14C]lysine. The large dose of leucine decreased FMPS rate by ~50% because acid-soluble muscle lysine (precursor) specific activity increased while the rate of incorporation of labeled lysine into protein decreased. These results indicate that the flooding dose method gives FMPS rates that are markedly greater than those obtained with the constant infusion method and that the latter method is not valid when a large dose of leucine is given.