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Measurement of Protein Turnover in Skeletal Muscle Strips^1,2,

Michigan State University⁴, East Lansing 48824-1225

Abstract

Isolated porcine and bovine muscle strips were incubated in Krebs Ringer bicarbonate buffer to determine in vitro protein synthesis (PS) and protein degradation (PD) rates to validate the in vitro system for use with livestock species. The addition of 5x plasma concentrations of amino acids to the medium stimulated PS 30%. Addition of 3.5 mM leucine to a leucine-deficient buffer supplemented with amino acids decreased PD 37% and stimulated PS 24%. The addition of .1 U/ml insulin reduced PD 28% and increased PS 30%. Protein degradation was elevated in longitudinally split rat soleus and extensor digitorum longus muscles compared to their contralateral intact muscles. Muscle strips must be removed within 15 min of exsanguination because PD rates become greatly elevated thereafter. ATP concentrations declined during incubation, but the addition of ATP or creatine had no effect on either PD or PS. Neither PD nor PS was affected by the addition of transferrin, fetuin, ascorbate, dexamethasone or indomethacin to the incubation medium. However, muscle strips were sensitive to the addition of triiodothyronine (T₃); PD was increased up to 75% as T₃ concentration was increased, and PS rates doubled compared to controls. Serum from mature barrows or gilts had no effect on protein turnover, but the addition of 10% and 15% serum from boars increased both PD and PS. With fasted pigs a continual decline in PS occurred over 5 d, whereas PD was elevated at 3 d and then declined to rates comparable to the fed state after 5 d. These data suggest that the in vitro system has application for assessing relative changes that occur in vivo following nutritional, physiological and endocrinological manipulation.

¹ The authors gratefully acknowledge the technical assistance of Kati Loeffler, Mary Kraus, Adrianne Danzeisen, Sally Johnson, Robert Burnett and Tom Forton.

² Present address: Dept. of Anim. and Dairy Sci., Auburn University, Auburn, AL 36849-4201.

³ Michigan Agric. Exp. Sta. journal art. no. 12185.

⁴ Dept. of Anim. Sci.-Growth Biol. Group, and Dept. of Food Sci. and Human Nutr.

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