The effects of either low (25 pnollmin) or high (235 pnol/min) infusion of NH&l into the mesenteric vein for 5 d were determined on 0, consumption plus urea and amino acid transfers across the portaldrained viscera (PDV) and liver of young sheep. Kinetic transfers were followed by use of l5NH4C1 for 10 h on the fifth day with simultaneous infusion of [l-'3C]leucine to monitor amino acid oxidation.Neither PDV nor liver blood flow were affected by the additional NH, loading, although at the higher rate there was a trend for increased liver 0, consumption. N&-N extraction by the liver accounted for 64-70 % of urea-N synthesis and at the lower infusion rate the additional N required could be more than accounted for by hepatic removal of free amino acids. At the higher rate of NH, administration additional sources of N were apparently required to account fully for urea synthesis. Protein synthesis rates in the PDV and liver were unaffected by NH3 infusion but both whole-body (P < 0.05) and splanchnic tissue leucine oxidation were elevated at the higher rate of administration. Substantial synthesis of [15wglutamine occurred across the liver, particularly with the greater NH, supply, and enrichments exceeded considerably those of glutamate. The [15N]urea synthesized was predominantly as the single labelled, i.e. [14N15N], species. These various kinetic data are compatible with the action of ovine hepatic glutamate dehydrogenase (EC1.4.1.2) in periportal hepatocytes in the direction favouring glutamate deamhation. Glutamate synthesis and uptake is probably confined to the perivenous cells which do not synthesize urea. The implications of NH, detoxification to the energy and N metabolism of the ruminant are discussed.

The response in whole-body and splanchnic tissue mass and isotope amino acid transfers in both plasma and blood has been studied in sheep offered 800 g lucerne (Medicago sutivu) pellets/d. Amino acid mass transfers were quantified over a 4 h period, by arteric+venous procedures, across the portal-drained viscera (PDV) and liver on day 5 of an intravenous infusion of either vehicle or the methylated products, choline (0.5 g/d) plus creatine (1.0 g/d). Isotopic movements were monitored over the same period during a 10 h infusion of a mixture of U-13C-labelled amino acids obtained from hydrolysis of labelled algal cells. Sixteen amino acids were monitored by gas chromatography-mass spectrometry, with thirteen of these analysed within a single chromatographic analysis. Except for methionine, which is discussed in a previous paper, no significant effects of choline plus creatine infusion were observed on any of the variables reported. Whole-body protein irreversible-loss rates ranged from 158 to 245 g/d for the essential amino acids, based on the relative enrichments (dilution of the U-I3C molecules by those unlabelled) of free amino acids in arterial plasma, and 206-519 g/d, when blood free amino acid relative enrichments were used for the calculations. Closer agreement was obtained between lysine, threonine, phenylalanine and the branched-chain amino acids. Plasma relative enrichments always exceeded those in blood (P < 0.001), possibly due to hydrolysis of peptides or degradation of protein within the erythrocyte or slow equilibration between plasma and the erythrocyte. Net absorbed amino acids across the PDV were carried predominantly in the plasma. Little evidence was obtained of any major and general involvement of the erythrocytes in the transport of free amino acids from the liver. Net isotope movements also supported these findings. Estimates of protein synthesis rates across the PDV tissues from [UJ3C] leucine kinetics showed good agreement with previous values obtained with single-labelled leucine. Variable rates were obtained between the essential amino acids, probably due to different intracellular dilutions. Isotope dilution across the liver was small and could be attributed predominantly to uni-directional transfer from extracellular sources into the hepatocytes and this probably dominates the turnover of the intracellular hepatic amino acid pools.Stable isotopes: Amino acids: Protein metabolism: Liver

~ ~~~Whole-body protein synthesis, estimated by the irreversible loss rate procedure, and hind-leg protein metabolism determined by arterio-venous techniques were monitored in response to three nutritional conditions (approximately 0.6, 1 2 and 1.8 x energy maintenance (M)) in ten wether lambs (33 kg average live weight). In all lambs and treatments measurements were based on radiolabelled phenylalanine, but the terminal procedures (five a t 0.6 x M and five at 1.8 x M) also included infusion of[ I-'3Clleucine ; this permitted comparison of amino acids catabolized (leucine) and non-metabolized (phenylalanine) by the hind-limb tissues. Whole-body protein synthesis increased with intake and the relationship with energy expenditure was slightly lower than that reported previously for pigs and cattle. The efficiency of protein retention: protein synthesis did not exceed 0.25 between the two intake extremes. Effects of intake on amino acid oxidation were similar to those observed for cattle. Hind-limb protein synthesis also increased significantly (P < 0001) in response to intake. Estimates of protein gain, from net uptake values, indicated that the tissues made a greater proportional contribution to total protein retention above M and to protein loss below M, emphasizing the role played by muscle tissue in providing mobile protein stores. The rates of protein synthesis calculated depended on the selection of precursor (blood) metabolite, but rates based on leucine always exceeded those based on phenylalanine when precursor from the same pool was selected. The incremental efficiency of protein retained: protein synthesis was apparently unity between 0.6 and 1.2 x M but 0.3 from 1.2 to 1.8 x M. Blood flow through the iliac artery was also proportional to intake. Leucine and 0x0-acid catabolism to carbon dioxide increased with intake such that the metabolic fate of the amino acid was distributed in the proportion 2: I between protein gain and oxidation. The rates of oxidation were only 1-3% the reported capacity of the rate-limiting dehydrogenase enzyme in muscle, but sufficient enzyme activity resides in the hindlimb adipose tissue to account for such catabolism. 1987) and, thus, small changes in the balance of the two processes can produce marked effects on net anabolism and production efficiency. One effective modulator of protein turnover is intake and for a variety of commercial species, including pig (Reeds et ul. 1980),

Rates of protein synthesis for the liver, plasma albumin and total plasma protein were quantified in sheep either offered a supra-maintenance intake or fasted for 3 d. The technique of continuous infusion over a 12 h period was employed with the simultaneous infusion of [l-'3C]glycine, phenylalanine. Blood and plasma samples were removed at timed intervals from the hepatic portal and hepatic veins plus the aorta. Enrichments of the free amino acids (AA) were determined in all blood and plasma samples as was the protein-bound AA in an apolipoprotein BlOO extract. Protein-bound phenylalanine enrichments were also measured in albumin and total protein from plasma plus samples from liver biopsies. The apolipoprotein BlOO enrichments agreed well with those of the free AA in hepatic (and hepatic portal) plasma but were lower than for arterial free AA and greater than liver homogenate free AA. This adds support to the concept that export proteins may preferentially use AA directly from extracellular sources. Intake had no significant effect on constitutive liver protein synthesis and the values agreed well with those obtained by other isotopic approaches. There were, however, sigdlicant declines, based on hepatic venous free phenylalanine enrichment, at the lower intake in both the fractional (3.4 v. 4.7 % per d; P = 0.024) and absolute (2.4 v. 4.2 g/d; P=O-O11) synthesis rates of albumin, which matched the estimated decrease in total plasma albumin content (52 v. 67 g, P < 0.01). In contrast, there was a smaller reduction in total plasma protein mass (145 Y. 151 g, P=O.O35) with no observed significant ditrerence in kinetic parameters. Albumin synthesis was calculated to account for a maximum of 17 % of total liver protein synthesis in the fed condition and this may fall to 8 % during moderate fasts.

In ten lambs (average live weight 33 kg), five offered 300 g/d (approximately 0 6 x maintenance; L) and five 900 g/d (1.8 x maintenance; H), tissue protein synthesis was measured by three procedures simultaneously. The techniques involved continuous infusion of leucine over 7-8 h followed by a terminal large dose of 115Nlphenylalanine during the last 30 or 60 min. Rates of protein synthesis were then calculated based on the free amino acid or 0x0-acid isotopic activity in either arterial, iliac venous blood or tissue homogenate for the continuous-infusion studies, or on plasma or tissue homogenate for the large-dose procedure. For muscle ( > 99 YO), and to a lesser extent skin amino or 0x0-acid were significantly less, more so at the lower intake. In contrast, for skin, a tissue dominated by export protein synthesis, values from the large-dose procedure (L 6.37 %/d, H 10.98 %Id) were similar to those derived with arterial or venous metabolites as precursor (L 5.23 and 693%/d, H 9.98 and 11.71 %/d for leucine), but much less than those based on homogenate data. Based on the large-dose technique, protein synthesis increased with intake in muscle (P < 0.001), skin (P = 0.009) and liver (26.7 v. 30.5 %/d; P = 0.029). The contributions of muscle and skin to total protein synthesis were approximately equal. The incremental efficiency of conversion for muscle of synthesized protein into deposition appeared to be similar to values reported for rodents.Tissue protein synthesis: Protein intake: LambThe dynamic nature of protein metabolism has been much investigated over the past two decades, with particular emphasis on understanding responses in tissues to a variety of nutritional and physiological stimuli. In both laboratory and commercial species most attention has focused on measurement of protein synthesis, for which a range of tracerbased techniques are available (see Lobley, 1988). Data for the larger species are more limited than for rodents, due to both accessibility and cost. Furthermore, much of the information on protein metabolism in individual tissues of farm animals is based on the continuous infusion of tracer amino acid developed by Waterlow and his colleagues (e.g. Garlick et al. 1973). The problem with this technique is that the various free amino acid pools of the body (e.g. vascular, interstitial, intracellular) become labelled to different