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Hindlimb protein turnover and muscle protein synthesis in lambs: a comparison of techniques

Published online by Cambridge University Press:  09 March 2007

L. A. Crompton
Affiliation:
Growth Biochemistry Group, Department of Biochemistry and Physiology, University of Reading, Whiteknights, PO Box 228, Reading, Berkshire, RG6 2AJ
M. A. Lomax
Affiliation:
Growth Biochemistry Group, Department of Biochemistry and Physiology, University of Reading, Whiteknights, PO Box 228, Reading, Berkshire, RG6 2AJ
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Abstract

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A combination of arterio–venous difference, kinetic isotope transfer and blood flow rate techniques were used to measure tyrosine metabolism across hindlimb tissues of nine growing lambs (average live weight 36.5 kg) fed on a range of dry matter intakes. Muscle protein synthesis was measured using a continuous infusion technique and compared with simultaneous estimates of hindlimb protein turnover calculated from the values for tyrosine metabolism. When the specific radioactivity (SRA) of tyrosine in the arterial plasma free pool was assumed to be the same as the SRA of tyrosine in the direct precursor pool of protein synthesis, hindlimb protein synthesis (ksav; 3.66 (SEM 0.50) %/d) was significantly (P < 0.001) higher (68%) than muscle protein synthesis (ksp; 2.18 (SEM 0.31) %/d) but was similar to the value for muscle protein synthesis calculated using the homogenate free tyrosine SRA (ksh; 3.35 (SEM 0.42) %/d). Hindlimb and muscle protein synthesis (y) were both significantly related to dry matter intake (x) (ksav, r2 0.667, P = 0.007; ksh, r2 0.968, P < 0.001) and there was no significant difference between the slopes (P = 0.532) and intercepts (P = 0.945) of the two regression lines. The results demonstrate that hindlimb protein turnover cannot be quantitatively compared with muscle protein synthesis, probably due to high protein metabolic activity in non-muscular tissues within the hindlimb, although similar responses in protein synthetic rate to the level of feed intake were observed between hindlimb and muscle tissues.

Type
Protein Metabolism
Copyright
Copyright © The Nutrition Society 1993

References

REFERENCES

Abdul-Razzaq, H. A. & Bickerstaffe, R. (1989). The influence of rumen volatile fatty acids on protein metabolism in growing lambs. British Journal of Nutrition 62, 297310.Google Scholar
Airhart, J., Arnold, J. A., Stirewalt, W. S. & Low, R. B. (1982). Insulin stimulation of protein synthesis in cultured skeletal and cardiac muscle cells. American Journal of Physiology 243, C81C86.CrossRefGoogle ScholarPubMed
Airhart, J., Vidrich, A. & Khairallah, E. A. (1974). Compartmentation of free amino acids for protein synthesis in rat liver. Biochemical Journal 140, 539548.CrossRefGoogle ScholarPubMed
Baird, G. D., Van Der Walt, J. G. & Bergman, E. N. (1983). Whole-body metabolism of glucose and lactate in productive sheep and cows. British Journal of Nutrition 50, 249265.CrossRefGoogle ScholarPubMed
Barrett, E. J. & Gelfand, R. A. (1989). The in vivo study of cardiac and skeletal muscle protein turnover. Diabetes Metabolism Reviews 5, 133148.Google Scholar
Barrett, E. J., Revkin, J. H., Young, L. H., Zaret, B. L., Jacob, R. & Gelfand, R. A. (1987). An isotopic method for measurement of muscle protein synthesis and degradation in vivo. Biochemical Journal 245, 223228.Google Scholar
Bird, A. R., Chandler, K. D. & Bell, A. W. (1981). Effects of exercise and plane of nutrition on nutrient utilisation by the hind-limb of sheep. Australian Journal of Biologica1 Sciences 34, 541550.Google Scholar
Bohorov, O., Buttery, P. J., Correia, J. H. R. D. & Soar, J. B. (1987). The effect of the β-2-adrenergic agonist clenbuterol or implantation with oestradiol plus trenbolone acetate on protein metabolism in wether lambs. British Journal of Nutrition 51, 99107.CrossRefGoogle Scholar
Boisclair, Y., Bauman, D. E., Bell, A. W. & Dunshea, F. R. (1988). Muscle protein synthesis and whole body N balance in fed and underfed steers. FASEB Journal 2, A848.Google Scholar
Bryant, D. T. W. & Smith, R. W. (1982). Protein synthesis in muscle of mature sheep. Journal of Agricultural Science, Cambridge 98, 639643.CrossRefGoogle Scholar
Chambers, J. A. & Bickerstaffe, R. (1982). Effect of rumen development on protein synthesis in lambs. Proceedings of the Nutrition Society of Australia 7, 148.Google Scholar
Cheng, K. N., Dworzak, F., Ford, G. C., Rennie, M. J. & Halliday, D. (1985). Direct determination of leucine metabolism and protein breakdown in humans using L-[1-13C,15N]-leucine and the forearm model. European Journal of Clinical Investigation 15, 349354.Google Scholar
Cheng, K. N., Pacy, P. J., Dworzak, F., Ford, G. C. & Halliday, D. (1987). Influence of fasting on leucine and muscle protein metabolism across the forearm determined using L-[1-13C,15N]-leucine as the tracer. Clinical Science 73, 241246.CrossRefGoogle ScholarPubMed
Cheng, K. N., Pacy, P. J., Hicks, C., Ford, G. C., Merritt, H. & Halliday, D. (1988). A non-invasive technique for determining muscle protein synthesis. Proceedings of the Nutrition Society 41, 54A.Google Scholar
Coufalik, A. & Monder, C. (1978). Perinatal development of the tyrosine oxidising system. Biology of the Neonate 34, 161166.Google Scholar
Crompton, L. A. (1990). Acute nutritional signals in the control of hind-limb protein turnover in lambs in vivo. PhD Thesis, University of Reading.Google Scholar
Davis, S. R., Barry, T. N. & Hughson, G. A. (1981). Protein synthesis in tissues of growing lambs. British Journal of Nutrition 46, 409419.Google Scholar
Domanski, A., Lindsay, D. B. & Setchell, B. P. (1974). Blood flow and substrate uptake and oxidation in the hindlimb muscles of sheep. Journal of Physiology 242, 28P29P.Google Scholar
Garlick, P. J. & Lobley, G. E. (1987). Dietary intake and protein turnover. In Protein Metabolism and Nutrition. European Association of Animal Production, Publication no. 35, pp. 1821, Rostock, Germany: Wilhelm-Pieck- Universität.Google Scholar
Garlick, P. J., McNurlan, M. A. & Preedy, V. R. (1980). A rapid and convenient technique for measuring the rate of protein synthesis in tissues by injection of [3H]phenylalanine. Biochemical Journal 192, 719723.Google Scholar
Garlick, P. J. & Marshall, I. (1972). A technique for measuring brain protein synthesis. Journal of Neurochemistry 19, 577583.CrossRefGoogle ScholarPubMed
Garlick, P. J., Millward, D. J. & James, W. P. T. (1973). The diurnal response of muscle and liver protein synthesis in vivo in meal-fed rats. Biochemical Journal 136, 935945.Google Scholar
Gelfand, R. A. & Barrett, E. J. (1987). Effect of physiologic hyperinsulinemia on skeletal muscle protein synthesis and breakdown in man. Journal of Clinical Invesligation 80, 16.Google Scholar
Goldberg, A. L., Tischler, M., DeMartino, G. & Griffin, G. (1980). Hormonal regulation of protein degradation and synthesis in skeletal muscle. Federation Proceedings 39, 3136.Google Scholar
Granner, D. K. & Hargrove, J. L. (1983). Regulation of the synthesis of tyrosine aminotransferase: the relationship to mRNATAT. Molecular and Cellular Biochemistry 53/54, 113128.Google ScholarPubMed
Halliday, D., Pacy, P. J., Cheng, K. N., Dworzak, F., Gibson, J. N. & Rennie, M. J. (1988). Rate of protein synthesis in skeletal muscle of normal man and patients with muscular dystrophy: a reassessment. Clinical Science 74, 237240.CrossRefGoogle ScholarPubMed
Harris, C. I. & Milne, G. (1977). The unreliability of urinary 3-methylhistidine excretion as a measure of muscle protein degradation in sheep. Proceedings of the Nutrition Society 35, 138A.Google Scholar
Harris, C. I. & Milne, G. (1980). The urinary excretion of N-methylhistidine in sheep: an invalid index of muscle protein breakdown. British Journal of Nutrition 44, 129140.CrossRefGoogle Scholar
Harris, P. M., Dellow, D. W. & Sinclair, B. R. (1989 a). Preliminary ‘in vivo’ measurements of protein and energy metabolism in the skin of sheep. Australian Journal of Agricultural Science 40, 879888.Google Scholar
Harris, P. M., Garlick, P. J. & Lobley, G. E. (1989 b). Interactions between energy and protein metabolism in the whole body and hind-limb of sheep in response to intake. In Energy Metabolism of Farm Animals, European Association of Animal Production, Publication no. 43, pp. 167170 [van der Honing, Y. and Close, W. H., editors]. Wageningen, The Netherlands: Pudoc.Google Scholar
Harris, P. M. & Lobley, G. E. (1991). Amino acid and energy metabolism in the peripheral tissues of ruminants. In Physiological Aspects of Digestion and Metubolism in Ruminants, pp. 201230 [Tsuda, T., Sasaki, Y. and Kawashima, R., editors]. London: Academic Press.Google Scholar
Holman, R. B. & Snape, B. M. (1983). Determination of L-tyrosine in rat brain by reverse-phase liquid chromatography with electrochemical detection. Journal of Chromatography 262, 415419.CrossRefGoogle Scholar
Horber, F. F., Horber-Feyder, C. M., Krayer, S., Schwenk, W. F. & Haymond, M. W. (1989). Plasma reciprocal pool specific activity predicts that of intracellular free leucine for protein synthesis. American Journal of Physiology 251, E385E399.Google Scholar
Lobley, G. E., Connell, A., Milne, E., Buchan, V., Calder, A. G., Anderson, S. E. & Vint, H. (1990). Muscle protein synthesis in response to testosterone administration in wether lambs. British Journal of Nutrition 64, 691704.Google Scholar
Lobley, G. E., Milne, V., Lovie, J. M., Reeds, P. J. & Pennie, K. (1980). Whole body and tissue protein synthesis in cattle. British Journal of Nutrition 43, 491502.CrossRefGoogle ScholarPubMed
Matthews, D. E., Schwarz, H. P., Yang, R. D., Motil, K. J., Young, V. R. & Bier, D. M. (1982). Relationship of plasma leucine and α-ketoisocaproate during a L-[l-13C]leucine infusion in man: a method for measuring human intracellular leucine tracer enrichment. Metabolism 31, 11051112.Google Scholar
Ministry of Agriculture, Fisheries and Food (1984). Energy Allowances and Feeding Systems for Ruminants. Reference Book no. 433. London: H. M. Stationery Office.Google Scholar
Oddy, V. H., Brown, B. W. & Jones, A. W. (1981). Measurement of organ blood flow using tritiated water. I. Hind-limb muscle blood flow in conscious ewes. Australian Journal of Biological Sciences 34, 419425.CrossRefGoogle Scholar
Oddy, V. H., Gooden, J. M. & Annison, E. F. (1984). Partitioning of nutrients in Merino ewes. I. Contribution of skeletal muscle, the pregnant uterus and the lactating mammary gland to total energy expenditure. Australian Journal of Biological Sciences 31, 375388.CrossRefGoogle Scholar
Oddy, V. H., Jones, A. W. & Warren, H. M. (1988). Phenylalanine as a marker of muscle protein synthesis. Proceedings of the Nutrition Society of Australia 13, 119.Google Scholar
Oddy, V. H. & Lindsay, D. B. (1986). Determination of rates of protein synthesis, gain and degradation in intact hind-limb muscle of lambs. Biochemical Journal 233, 417425.CrossRefGoogle ScholarPubMed
Oddy, V. H., Lindsay, D. B., Barker, P. J. & Northrop, A. J. (1987). Effect of insulin on hind-limb and whole body leucine and protein metabolism in fed and fasted lambs. British Journal of Nutrition 58, 437452.Google Scholar
Ohisalo, J. J., Anderson, B. M., Viljanen, A. A. & Anderson, S. M. (1982). Is there a brain tyrosine aminotransferase? Biochemical Journal 204, 621622.Google Scholar
Pacy, P. J., Cheng, K. N., Rennie, M. J. & Halliday, D. (1988). Muscle protein synthesis rate – a reappraisal of control values. Proceedings of the Nutrition Society 41, 53A.Google Scholar
Pappenheimer, J. R. & Setchell, B. P. (1972). The measurement of cerebral blood flow in the rabbit and sheep. Journal of Physiology 226, 48P50P.Google Scholar
Pell, J. M., Caldarone, E. M. & Bergman, E. N. (1986). Leucine and α-ketoisocaproate metabolism and interconversions in fed and fasted sheep. Metabolism 35, 10051016.CrossRefGoogle ScholarPubMed
Pethick, D. W., Harman, N. & Chong, J. K. (1987). Non-esterified long chain fatty acid metabolism in fed sheep at rest and during exercise. Australian Journal of Biological Sciences 40, 221234.Google Scholar
Preedy, V. R., McNurlan, M. A. & Garlick, P. J. (1983). Protein synthesis in skin and bone of the young rat. British Journal of Nutrition 49, 517523.Google Scholar
Seve, B., Reeds, P. J., Fuller, M. F., Cadenhead, A. & Hay, S. M. (1986). Protein synthesis and retention in some tissues of the young pig as influenced by dietary protein intake after early weaning. Possible connection to the energy metabolism. Reproduction, Nutrition, Developpement 26, 849861.CrossRefGoogle Scholar
Teleni, E. & Annison, E. F. (1986). Development of a sheep hind-limb muscle preparation for metabolic studies. Australian Journul of Biological Sciences 39, 271281.CrossRefGoogle ScholarPubMed
Teleni, E., Annison, E. F. & Lindsay, D. B. (1986). Metabolism of valine and the exchange of amino acids across the hind-limb muscles of fed and starved sheep. Australian Journal of Biological Sciences 39, 379393.Google Scholar
Thompson, G. N., Pacy, P. J., Merritt, H., Ford, G. C., Read, M. A., Cheng, K. N. & Halliday, D. (1989). Rapid measurement of whole body and forearm protein turnover using a [2H5]phenylalanine model. American Journal of Physiology 256, E631E639.Google Scholar
Tomas, F. M. & Ballard, F. J. (1987). Applications of the N-methylhistidine technique for measuring myofibrillar protein breakdown in vivo. In Lysosomes: Their Role in Protein Breakdown, pp. 679711 [Glaumann, H. and Ballard, F. J., editors]. London: Academic Press.Google Scholar
Vincent, R. (1984). Protein metabolism in sheep: measurement in vivo and the effects of pregnancy and lactation. PhD Thesis, University of Cambridge.Google Scholar
Waalkes, T. P. & Udenfriend, S. (1957). A fluorometric method for the estimation of tyrosine in plasma and tissues. Journal of Laboratory and Clinical Medicine 50, 773–736.Google Scholar
Waterlow, J. C., Garlick, P. J. & Millward, D. J. (1978). Protein Turnover in Mammalian Tissues and in the Whole Body. London: North-Holland.Google Scholar