Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-20T11:46:07.258Z Has data issue: false hasContentIssue false
  • English
  • Français

Whole body metabolism of cysteine and glutathione and their utilization in the skin of Romney sheep: consequences for wool growth

Published online by Cambridge University Press:  27 March 2009

J. Lee ,
P. M. Harris ,
B. R. Sinclair  and
B. P. Treloar
J. Lee
Affiliation:
AgResearch Grasslands, Private Bag 11008, Palmerston North, New Zealand
P. M. Harris
Affiliation:
AgResearch Grasslands, Private Bag 11008, Palmerston North, New Zealand
B. R. Sinclair
Affiliation:
AgResearch Grasslands, Private Bag 11008, Palmerston North, New Zealand
B. P. Treloar
Affiliation:
AgResearch Grasslands, Private Bag 11008, Palmerston North, New Zealand
Get access Rights & Permissions [Opens in a new window]

Summary

Infusion of [35S]-labelled cysteine into the jugular of Romney sheep of different ages was used to estimate the extent of combined cysteine and cystine (cyst(e)ine) and glutathione (GSH) interchange in various tissues, including skin, and to measure whole body irreversible loss rates (1LR) for cyst(e)ine. The studies were undertaken at Palmerston North, New Zealand during 1990 and 1991.

Reverse phase HPLC using a fluorigenic reagent, 7-fluoro-2,1,3-benzoxadiazole-4-sulphonate (SBD-F) was used for the simultaneous determination of thiols (cysteine and GSH) in whole blood (bound and free fractions), liver, kidney, small intestine, muscle, pancreas and skin. The appearance of [35S]-label in thiol compounds and their oxidation products was determined by ion-exchange HPLC. Specific radioactivities (SRA) for cyst(e)ine and GSH derived from this data showed equivalence for cysteine and GSH SRAs in all tissues, except for whole blood, indicating rapid withintissue interconversion between these thiols. In whole blood, however, the very low SRA for GSH (< 4 DPM/nmol) compared to cyst(e)ine (250 DPM/nmol) indicated markedly slower or negligible exchange with red blood cell GSH, and hence little inter-organ transport of [35S]-label as GSH.

Close infusion of [35S] cysteine into a defined patch of skin and collection of the venous outflow permitted direct in vivo measurement of the proportional uptake of cysteine by the skin. Results indicated considerable variation in the uptake of cysteine per se (20–40%) but no, or very little, oxidation of cysteine in the skin and no net export of GSH.

The combination of whole body, tissue and skin specific studies of [35S]-labelled cysteine metabolism quantitatively confirmed the very high proportion of circulating cyst(e)ine in sheep which is directed to skin and wool protein synthesis alone, and highlighted those aspects of this metabolism which are of most importance to wool production.

Type
Animals
Information
The Journal of Agricultural Science , Volume 121 , Issue 1 , August 1993 , pp. 111 - 124
Copyright
Copyright © Cambridge University Press 1993

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Akerboom, T. P. M. & Sies, H. (1989). Transport of glutathione, glutathione disulfide, and glutathione conjugates across the hepatocyte plasma membrane. Methods in Enzymology 173, 523–534.CrossRefGoogle ScholarPubMed
Anderson, N. E. & Meister, A. (1987). Intracellular delivery of cysteine. Methods in Enzymology 143, 313–325.CrossRefGoogle ScholarPubMed
Benevenga, N. J., Radcliffe, B. C. & Egan, A. R. (1983). Tissue metabolism of methionine in sheep. Australian Journal of Biological Sciences 36, 475–485.CrossRefGoogle ScholarPubMed
Black, J. L. & Reis, P. J. (1979). Speculation on the control of nutrient partition between wool growth and other body functions. In Physiological and Environmental Limitations to Wool Growth (Eds Black, J. L. & Reis, P. J.), pp. 269–293. Armidale, Australia: University of New England.Google Scholar
Bremmers, R. P. M., Morgan, P. F., McCutcheon, S. N. & Purchas, R. W. (1988). Effect of plane of nutrition on energy and nitrogen retention and on plasma urea concentrations in Southdown ram hoggets from high and low backfat selection lines. New Zealand Journal of Agricultural Research 31, 1–7.CrossRefGoogle Scholar
Camas, H., Baspinar, H., Antapli, M., Ogan, C. & Sener, E. (1986). Relationship of serum glutathione concentration with fleece traits in Ramlic yearling ewes. Animal Breeding Abstracts (1990) 058–01527.Google Scholar
Clark, C. M., Mackenzie, D. D. S., McCutcheon, S. N. & Blair, H. T. (1989). Physiological responses to selection for greasy fleeceweight in Romney sheep. New Zealand Journal of Agricultural Research 32, 29–36.CrossRefGoogle Scholar
Cobon, D. H., Suter, G. R., Connelly, P. T., Shepherd, R. K. & Hopkins, P. S. (1988). The residual effects of methionine supplementation on the wool growth performance of grazing sheep. Proceedings of the Australian Society of Animal Production 17, 383.Google Scholar
D'andrea, R. J., Sivaprassad, A. V., Bawden, S., Kuczek, E. S., Whitbread, L. A. & Rogers, G. E. (1989). Isolation of microbial genes for cysteine synthesis and prospects for their use in increasing wool growth. In The Biology of Wool and Hair (Eds Rogers, G. E., Reis, P. J., Ward, K. A., & Marshall, R. C.), pp. 447–463. London: Chapman and Hall.Google Scholar
Deneke, S. M. & Fanburg, B. L. (1989). Regulation of cellular glutathione. American Journal of Physiology 257, L163–L173.Google ScholarPubMed
Dimant, E., Landsberg, E. & London, I. M. (1955). The metabolic behaviour of reduced glutathione in human and avian erythrocytes. Journal of Biological Chemistry 213, 769–776.CrossRefGoogle ScholarPubMed
Fenn, P. D. & Leng, R. A. (1989). Wool growth and sulfur amino acid entry rate in sheep fed roughage based diets supplemented with bentonite and sulfur amino acids. Australian Journal of Agricultural Research 40, 889–896.CrossRefGoogle Scholar
Flebrand, G., Ozapo, V., Wassmuth, R. & Freudenreich, P. (1989). Association of blood concentration of glutathione with meat and fat gain and fitness in various sheep breeds and crosses. Journal of Animal Breeding and Genetics 106, 59–66.Google Scholar
Gaitonde, M. K. (1967). A spectrophotometric method for the direct determination of cysteine in the presence of other naturally occurring amino acids. Biochemical Journal 104, 627–633.CrossRefGoogle ScholarPubMed
Garlick, P. J. (1980). Assessment of protein metabolism in the intact animal. In Protein Deposition in Animals (Eds Buttery, P. J. & Lindsay, D. B.), pp. 51–68. London: Butterworths.CrossRefGoogle Scholar
Harris, P. M., Dellow, D. W. & Sinclair, B. R. (1989). Preliminary ‘in vivo’ measurements of protein and energy metabolism in the skin of sheep. Australian Journal of Agricultural Research 40, 879–888.CrossRefGoogle Scholar
Hoey, W. A., Broadmeadow, A. C. C. & Hopkins, P. S. (1984). A study of the peripheral utilisation of sulphur amino acids for fibre production in the sheep. Proceedings of the Australian Society of Animal Production 15, 120–123.Google Scholar
Hopkins, G. J., Roberts, J. & Evans, J. V. (1975). Red blood cell concentrations of reduced glutathione and potassium as biological markers of wool growth in Merino sheep. Journal of Agricultural Science, Cambridge 84, 481–486.CrossRefGoogle Scholar
Lee, P. L. Y. & Slocum, R. H. (1988). A high resolution method for amino acid analysis of physiological fluids containing mixed disulphides. Clinical Chemistry 34, 719–723.CrossRefGoogle Scholar
Meister, A. (1973). On the enzymology of amino acid transport. Science 180, 33–39.CrossRefGoogle ScholarPubMed
Meister, A. & Anderson, M. E. (1983). Glutathione. Annual Review of Biochemistry 52, 711–760.CrossRefGoogle ScholarPubMed
Pisulewski, P. M. & Buttery, P. J. (1985). The effect of increasing methionine supply on the methionine conversion to cyst(e)ine in sheep. British Journal of Nutrition 54, 121–129.CrossRefGoogle ScholarPubMed
Radcliffe, B. C. & Egan, A. R. (1978). The effect of diet and of methionine loading on activity of enzymes in the transulfuration pathway in sheep. Australian Journal of Biological Sciences 31, 105–114.CrossRefGoogle ScholarPubMed
Reeds, P. J. (1992). Isotopic estimation of protein systhesis and proteolysis. In Modern Methods in Protein Nutrition and Metabolism (Ed. Nissen, S.), pp. 249–273. London: Academic Press.CrossRefGoogle Scholar
Reis, P. J. (1986). The relative efficiency of methionine, cysteine, and homocysteine for promoting wool growth. Proceedings of the Nutrition Society of Australia II, 166–170.Google Scholar
Reis, P. J., Tunks, D. A. & Sharry, L. F. (1989). Incorporation of abomasal and intravenous doses of [35S]cystine and [35S]methionine into wool. Journal of Agricultural Science, Cambridge 112, 313–319.CrossRefGoogle Scholar
Reis, P. J., Tunks, D. A. & Munro, S. G. (1990). Effects of the infusion of amino acids into the abomasum of sheep, with emphasis on the relative value of methionine, cysteine and homocysteine for wool growth. Journal of Agricultural Science, Cambridge 114, 59–68.CrossRefGoogle Scholar
Rizzi, R., Caroli, A., Bolla, P., Acciaioli, A. & Pagnacco, G. (1988). Variability of reduced glutathione levels in Massese ewes and its effect on daily milk production. Journal of Dairy Research 55, 345–353.CrossRefGoogle ScholarPubMed
Sandholm, M., Atroshi, F. & Sankari, S. (1983). Erythrocyte GSH as carrier of amino acids into the lactating mammary gland – a study in sheep and goats. In Production Disease in Farm Animals, 5th International Conference, Uppsala, Sweden (1983), pp. 81–84.Google Scholar
Standeven, A. M. & Wetterhahn, K. E. (1991). Tissuespecific changes in glutathione and cysteine after buthionine sulfoximine treatment of rats and the potential for artifacts in thiol levels resulting from tissue preparation. Toxicology and Applied Pharamacology 107, 269–284.CrossRefGoogle ScholarPubMed
Tagari, H. & Bergman, E. N. (1978). Intestinal disappearance and portal blood appearance of amino acids in sheep. Journal of Nutrition 108, 790–803.CrossRefGoogle ScholarPubMed
Toyo'oka, T., Uchiyama, S. & Saito, Y. (1988). Simultaneous determination of thiols and disulphides by high performance liquid chromatography with fluorescence detection. Analytica Chimica Ada 205, 29–41.CrossRefGoogle Scholar
Viña, J. R., Palacin, M., Puertes, I. R., Hernandez, R. & Viña, J. (1989). Role of the γ-glutamyl cycle in the regulation of amino acid translocation. American Journal of Physiology 257, E916–E922.Google ScholarPubMed
Williams, A. J. (1976). Metabolism of cystine by Merino sheep genetically different in wool production. IV. Rates of entry of cystine into plasma, measured with a single intravenous injection of L-[35S]cystine, and the subsequent incorporation of 35S into wool fibres. Australian Journal of Biological Sciences 29, 513–524.CrossRefGoogle ScholarPubMed
Williams, A. J. (1987). Physiological consequences of selection for increased fleeceweight. In Merino Improvement Programs in Australia (Ed. McGuirk, B. J.), pp. 481–494. Sydney: Australian Wool Corporation.Google Scholar
Williams, A. J., Leng, R. A. & Stephenson, S. K. (1972). Metabolism of cystine by Merino sheep genetically different in wool production. I. Comparison of the entry rates of cystine in sheep from flocks selectively bred for high and low fleece weight. Australian Journal of Biological Sciences 25, 1259–1268.CrossRefGoogle ScholarPubMed
Williams, A. J., Murison, R. D. & Cross, C. (1986). Reactions of cyst(e)ine concentration in plasma of Merino sheep from two genetic groups to fasting and ACTH injection. Australian Journal of Agricultural Research 37, 657–663.CrossRefGoogle Scholar
Williams, A. J., Thornberry, K. J. & Gilmour, A. R. (1990). The activity of the thyroid glands in relation to genetic differences in wool production. Australian Journal of Agricultural Research 41, 573–581.CrossRefGoogle Scholar