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.