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Metabbolism of perfused ruminant muscle

Published online by Cambridge University Press:  27 March 2009

B. Jane Coward
Affiliation:
Department of Applied Biochemistry and Nutrition, University of Nottingham School of Agriculture, Sutton Bonington, Loughborough, LE12 5RD
P. J. Buttery
Affiliation:
Department of Applied Biochemistry and Nutrition, University of Nottingham School of Agriculture, Sutton Bonington, Loughborough, LE12 5RD

Summary

Substrate and nitrogen metabolism of ruminant muscle was studied in vitro using a perfused sheep hemidiaphragm preparation. Glucose, acetate, propionate and butyrate were taken up by the perfused muscle at a constant rate over 2 h. Rate of acetoacetate uptake declined over the perfusion period. Net production of 3-hydroxybutyrate over the perfusion period was observed.

Release of alanine and glycine by the perfused muscle could not be accounted for by protein breakdown, implicating muscle as a site of synthesis of these amino acids. The perfused muscle metabolized aspartate and arginine to a considerable extent, but the branched-chain amino acids only to a small extent. Only a small proportion, approximately 5%, of the alanine produced by the perfused muscle was derived from glucose.

In the presence of amino acids, at twice the concentration found in plasma, and insulin, some amino acids showed a net uptake by the diaphragm but when cysteine was omitted from the perfusate, all amino acids with the exception of arginine showed a net output. Evidence was obtained that protein synthesis was reduced in the absence of cysteine. The output of Nτ methylhistidine by the preparation was much higher than could be accounted for by estimates of protein breakdown using isotope dilution techniques. These data are, however, consistent with the view that Nτ methylhistidine is not a good index of muscle protein breakdown in sheep.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1982

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References

REFERENCES

Arnal, M. (1977). Muscle protein turnover in lambs throughout development. In Protein Metabolism and Nutrition. Proceedings 2nd International Symposium on Protein Metabolism and Nutrition, The Netherlands, pp. 3537. Pudoc Centre for Agricultural Publication and Documentation, Wageningen.Google Scholar
Atkin, G. E. & Ferdinand, W. (1970). Accelerated amino acid analysis: studies on the use of lithium citrate buffers and the effect of n-propanol in the analysis of physiological fluids and protein hydrolysates. Analytical Biochemistry 38, 313329.CrossRefGoogle Scholar
Ballard, F. J., Filsell, O. H. & Jarrett, I. G. (1976). Amino acid uptake and output by sheep hind limb. Metabolism: Clinical and Experimental 25, 415418.CrossRefGoogle ScholarPubMed
Beckerton, A. (1976). Protein turnover in sheep. Ph. D. thesis, University of Nottingham.Google Scholar
Beckerton, A., Buttery, P. J., Bailey, F. J. & Bolton, N. (1975). Improved method for the separation of lysine from N-ε-monomethyllysine in plasma using cation-exchange chromatography. Journal of Chromatography 104, 170171.CrossRefGoogle ScholarPubMed
Bell, A. W., Gardner, J. W., Manson, W. & Thompson, G. E. (1975). Acute cold exposure and the metabolism of blood glucose, lactate and pyruvate, and plasma amino acids in the hind limb of the fed and fasted young ox. British Journal of Nutrition 33, 207217.Google Scholar
Bell, A. W., Gardner, J. W. & Thompson, G. E. (1974). The effects of acute cold exposure and feeding of volatile fatty acid metabolism in the hind leg of the young ox. British Journal of Nutrition 32, 471477.Google Scholar
Bell, A. W. & Thompson, G. E. (1974). Effects, of cold exposure and feeding on net exchange of plasma free fatty acids and glycerol across the hind limb of the young ox. Research in Veterinary Science 17, 265267.CrossRefGoogle ScholarPubMed
Berger, M., Hagg, S. & Ruderman, N. B. (1975). Glucose metabolism in perfused skeletal muscle, interaction of insulin and exercise on glucose uptake. Biochemical Journal 146, 231238.CrossRefGoogle ScholarPubMed
Bergman, E. N. (1975). Production and utilization of metabolites by the alimentary tract as measured in portal and hepatic blood. In Digestion and Metabolism in the Ruminant. Proceedings IV International Symposium on Ruminant Physiology, Sydney, Australia (ed. McDonald, I. W. and Warner, A. C. I.), pp. 292305. The University of New England Publications Unit, Armidale, Australia.Google Scholar
Bergmeyer, H. U. & Bernt, E. (1974). D-glucose determinatiocn with glucose oxidase and peroxidase. In Methods of Enzymatic Analysis, 2nd English edn., vol. 3, (ed. Bergmeyer, H. U.), pp. 12051215, New York, San Francisco, London: Academic Press.Google Scholar
Boling, J. A., Willard, J. C. & Burris, W. R. (1977). Isolation – perfusion of ovine hind limbs. III. Perfusate lysine levels. Journal of Animal Science 45, 832836.CrossRefGoogle ScholarPubMed
Buttery, P. J., Beckerton, A., Mitchell, R. M.Davies, K. & Annison, E. F. (1975). The turnover rate of muscle and liver protein in sheep. Proceedings of the Nutrition Society 34, 91A.Google ScholarPubMed
Chalmers, M. I., Grant, I. & White, F. (1977). Free amino nitrogen levels across the hindquarters of sheep. Proceedings of the Nutrition Society 36, 123A.Google ScholarPubMed
Chang, T. W. & Goldberg, A. L. (1978). The origin of alanine production in skeletal muscle. Journal of Biological Chemistry 253, 36773684.Google Scholar
Coward, B.J. & Buttery, P. J. (1980). A perfused ruminant muscle preparation. Journal of Agricultural Science, Cambridge 95, 716.Google Scholar
Cross, D. L., Boling, J. A. & Ely, D. G. (1974). Isolation-perfusion of ovine hind limbs. II. Amino acid metabolism. Journal of Animal Science 39, 898902.CrossRefGoogle ScholarPubMed
Domanski, A., Lindsay, D. B. & Setchell, B. P. (1974). Blood flow and substrate uptake and oxidation in the hind limb muscles of sheep. Journal of Physiology 242, 28P29P.Google ScholarPubMed
Felig, P. (1973). The glucose-alanine cycle. Metabolism 22, 179207.Google Scholar
Felig, P. (1975). Amino acid metabolism in man. Annual Review of Biochemistry 44, 933955.CrossRefGoogle ScholarPubMed
Felig, P., Pozefsky, T., Marliss, E. & Cahill, G. F. Jr (1970). Alanine: key role in gluconeogenesis. Science, New York 167, 10031004.CrossRefGoogle ScholarPubMed
Fulks, K. M., Li, J. B. & Goldberg, A. L. (1975). Effect of insulin, glucose and amino acids on protein turnover in rat diaphragm. Journal of Biological Chemistry 250, 290298.CrossRefGoogle ScholarPubMed
Gale, E. F. (1974). L-amino acids, L-lysine, L-arginine, L-ornithine, L-tyrosine, L-histidine, L-glutamic acid, L-aspartic acid, manometric method. In Methods of Enzymatic Analysis, 2nd English edn., vol. 4, (ed. Bergmeyer, H. U.), pp. 16621668. New York, San Francisco, London: Academic Press.Google Scholar
Goldberg, A. L. & Dice, J. F. (1974). Intracellular protein degradation in mammalian and bacterial cells. Annual Review of Biochemistry 43, 835869.Google Scholar
Goodman, M. M., Berger, M. & Ruderman, N. B. (1974). Glucose metabolism in rat skeletal muscle at rest, effect of starvation, diabetes, ketone bodies and free fatty acids. Diabetes 23, 881888.CrossRefGoogle ScholarPubMed
Grubb, B. (1976). De novo synthesis of alanine by the perfused rat hind limb. American Journal of Physiology 230, 13791384.CrossRefGoogle Scholar
Gulli, R. & Searle, G. L. (1973). Alanine turnover in the human: determination of the specific activity. Proceedings of the Society for Experimental Biology and Medicine 144, 830833.CrossRefGoogle ScholarPubMed
Hamilton, P. B. & Van Slyke, D. D. (1943). The gasometric determination of free amino acids in blood filtrates by the ninhydrin-CO2 method. Journal of Biological Chemistry 150, 231250.CrossRefGoogle Scholar
Harris, C. I. & Milne, G. (1980). The urinary excretion of Nτ-methylhistidine in sheep – an invalid index of muscle protein breakdown. British Journal of Nutrition 44, 129140.CrossRefGoogle Scholar
Heitman, R. N. & Bergman, E. N. (1980). Integration of amino acid metabolism in sheep: effects of fasting and acidosis. American Journal of Physiology 239, E248E254.Google Scholar
Jarrett, I. G., Filsell, O. H. & Ballard, F. J. (1976). Utilization of oxidizable substrates by the sheep hind limb: effects of starvation and exercise. Metabolism 25, 523531.CrossRefGoogle ScholarPubMed
Jefferson, L. S., Koehler, J. O. & Morgan, H. E. (1972). Effect of insulin on protein synthesis in skeletal muscle of an isolated perfused pieparation of rat hemicorpus. Proceedings of the National Academy of Science U. S. A. 69, 816820.CrossRefGoogle ScholarPubMed
Jefferson, L. S., Li, J. B. & Rannels, S. R. (1977). Regulation by insulin of amino acid ielease and protein turnover in the perfused rat hemicorpus. Joiwnal of Biological Chemistry 252, 14761483.CrossRefGoogle ScholarPubMed
Jefferson, L. S., Rannels, D. E., Munger, B. L. & Morgan, H. E. (1974). Insulin in the regulation of protein turnover in heart and skeletal muscle. Federation Proceedings 33, 10981104.Google ScholarPubMed
Jones, G. B. (1965). Determination of the specific activity of labelled blood glucose by liquid scintillation using glucose pentaacetate. Analytical Biochemistry 12, 249258.CrossRefGoogle ScholarPubMed
Krebs, H. A. & Eggleston, L. V. (1945). Metabolism of acetoacetate in animal tissues. Biochemical Journal 39, 408419.CrossRefGoogle ScholarPubMed
Leng, R. A. & Annison, E. F. (1964). The metabolism of D(-)-β-hydroxybutyrate in sheep. Biochemical Journal 90, 464469.CrossRefGoogle Scholar
Lindsay, D. B., Steel, J. W. & Barker, P. J. (1977). Metabolism of alanine in sheep muscle. Proceedings of the Nutrition Society 36, 80A.Google Scholar
Lindsay, D. B., Steel, J. W. & Buttery, P. J. (1976). The output of amino acids from muscle of fed and starved sheep. Proceedings of the Nutrition Society 36, 33A.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
London, D. R., Foley, T. H. & Webb, C. G. (1965). Evidence for the release of individual amino acids from the resting human forearm. Nature, London 208, 588589.Google Scholar
Meister, A. (1965). Biochemistry of the Amino Adds, 2nd edn., vol. II. New York, London: Academic Press.Google Scholar
Mellanby, J. & Williamson, D. H. (1974). Acetoacetate. In Methods of Enzymatic Analysis, 2nd English edn., Vol. 4, (ed. Bergmeyer, H. U.), pp. 18401843. New York, San Francisco, London: Academic Press.CrossRefGoogle Scholar
Odessey, R., Khairallah, E. A. & Goldberg, A. L. (1974). Origin and possible significance of alanine production by skeletal muscle. Journal of Biological Chemistry 249, 76237629.CrossRefGoogle ScholarPubMed
Pozefsky, T., Felig, F., Tobin, J. D., Soeldner, J. S. & Cahill, G. F. Jun (1969). Amino acid balance across tissues of the forearm in postabsorptive man. Effects of insulin at two dose levels. Journal of Clinical Investigation 48, 22732282.CrossRefGoogle ScholarPubMed
Radcliffe, B. C. & Egan, A. R. (1974). A survey of methionine adenosyltransferase and cystathionine-γ-lyase activities in ruminant tissues. Australian Journal of Biological Science 27, 465471.CrossRefGoogle ScholarPubMed
Rannels, D. E., Hjalmarson, A. C. & Morgan, H. E. (1974). Effects of non-carbohydrate substances on protein synthesis in muscle. American Journal of Physiology 226, 528539.CrossRefGoogle Scholar
Rannels, D. E., Kao, R. & Morgan, H. E. (1975). Effect of insulin on protein turnover in heart muscle. Journal of Biological Ckmietry 250, 16941701.Google Scholar
Rannels, D. E., Kao, R. & Morgan, H. E. (1976). Effect of cardiac ischemia on protein degradation. Circulation 53 (Supplement I), 130133.Google ScholarPubMed
Ruderman, N. B. & Berger, M. (1974). The formation of glutamine and alanine in skeletal muscle. Journal of Biological Chemistry 249, 55005506.CrossRefGoogle ScholarPubMed
Ruderman, N. B. & Lund, P. (1972). Amino acid metabolism in skeletal muscle-regulation of glutamine and alanine release in the perfused rat hindquarter. Israeli Journal of Medical Science 8, 295302.Google ScholarPubMed
Soharrer, E. & Blatt, J. (1976). In-vitro-untersuchungen zur amino-saurenaufnahme in die leber und muskelzelle beim lamm. Zentralblatt für Veterinär Medizin 23, 121130.CrossRefGoogle Scholar
Scharrer, E. & Huntemann, H. (1977). Developmental changes of monosaccharide uptake into skeletal muscle of the lamb. Pflugers Archiv 369, 6164.Google Scholar
Somogyi, M. (1945). Determination of blood sugar. Journal of Biological Chemistry 160, 6973.CrossRefGoogle Scholar
Strohfeldt, P., Kettl, H. & Weinges, K. F. (1974). Perfusion of the isolated rat hind limb with a synthetic medium. Hormone and Metabolic Research 6, 167168.CrossRefGoogle ScholarPubMed
Sutton, J. D. & Johnson, V. W. (1969). Fermentation in the rumen of cows given rations containing hay and flaked maize or rolled barley in widely different proportions. Journal of Agricultural Science, Cambridge 73, 459468.CrossRefGoogle Scholar
Theinhaus, R., Tharandt, L., Zais, V. & Staib, W. (1975). Effect of glucocorticoids on the release of amino acids in the perfused rat hindquarter. Hoppe-Seyler's Zeitschrift für Physiologische Chemie 356, 811817.Google Scholar
Vernon, B. G. & Buttery, P. J. (1976). Protein turnover in rats treated with trenbolone acetate. British Journal of Nutrition 36, 575579.CrossRefGoogle Scholar
Ward, L. C. & Buttery, P. J. (1978). Nτ-methylhistidine as an index of myofibrillar protein breakdown – an appraisal. Life Sciences 23, 11031116.CrossRefGoogle ScholarPubMed
Williamson, J. R. & Krebs, H. A. (1961). Acetoacetate as fuel of respiration in the perfused rat heart. Biochemical Journal 80, 540547.Google Scholar
Williamson, J. R. & Mellanby, J. (1974). D-(-)-)-3-hydroxybutyrate. In Methods of Enzymatic Analysis 2nd English edn., vol. 4, (ed. Bergmeyer, H. U.) pp. 18361839. New York, San Francisco, London: Academic Press.CrossRefGoogle Scholar
Wolff, J. E., Bergman, E. N. & Williams, H. H. (1972). Net metabolism of plasma amino acids by liver and portal drained viscera of fed sheep. American Journal of Physiology 223, 455460.CrossRefGoogle ScholarPubMed