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. 2012:798:245-64.
doi: 10.1007/978-1-61779-343-1_14.

In vivo measurement of muscle protein synthesis rate using the flooding dose technique

Marta L Fiorotto  1 , Horacio A Sosa Jr, Teresa A Davis
Affiliations

In vivo measurement of muscle protein synthesis rate using the flooding dose technique

Marta L Fiorotto et al. Methods Mol Biol. 2012.

Abstract

Skeletal muscle mass is determined by the balance between rates of protein synthesis and degradation. Protein synthesis rates can be measured in vivo by administering an amino acid as a tracer that is labeled with an isotope (radioactive or stable) of C, H, or N. The rate at which the labeled amino acid is incorporated into muscle protein, as a function of the amount of labeled amino acid in the precursor pool at the site of translation, reflects the rate of protein synthesis. There are a number of approaches for performing this measurement depending on the question being addressed and the experimental system being studied. In this chapter, we describe the "flooding dose" approach using L-[(3)H]-phenylalanine as the tracer and that is suitable for determining the rate of skeletal muscle protein synthesis (total and myofibrillar proteins) over an acute period (ideally less than 30 min) in any size animal; details for working with mice are presented. The method describes how to administer the tracer without anesthesia, the tissue collection, and the preparation of muscle and blood samples for analysis of the tracer and tracee amino acids in the precursor pool and in muscle proteins.

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References

    1. Davis TA, Fiorotto ML. Regulation of muscle growth in neonates. Curr Opin Clin Nutr Metab Care. 2009;12:78–85. - PMC - PubMed
    1. Davis TA, Fiorotto ML. Regulation of skeletal muscle protein metabolism in growing animals. In: Burrin DG, Mersmann HJ, editors. Biology of Metabolism of Growing Animals. Elsevier; The Netherlands: 2005. pp. 37–68.
    1. Rooyackers OE, Nair KS. Hormonal regulation of human muscle protein metabolism. Annu Rev Nutr. 1997;17:457–485. - PubMed
    1. Davis TA, Fiorotto ML, Burrin DG, et al. Protein synthesis in organs and tissues: quantitative methods in laboratory animals. In: El-Khoury AE, editor. Methods for Investigation of Amino Acid and Protein Metabolism. CRC Press; Boca Raton: 1999. pp. 49–68.
    1. Reeds PJ, Davis TA. Of flux and flooding: the advantages and problems of different isotopic methods for quantifying protein turnover in vivo: I. Methods based on the dilution of a tracer. Curr Opin Clin Nutr Metab Care. 1999;2:23–28. - PubMed

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