cysteinylglycine and acivicin

Deficiency of the antioxidant glutathione in brain appears to be connected with several diseases characterized by neuronal loss. To study neuronal glutathione metabolism and metabolic interactions between neurons and astrocytes in this respect, neuron-rich primary cultures and transient cocultures of neurons and astroglial cells were used. Coincubation of neurons with astroglial cells resulted within 24 hr of incubation in a neuronal glutathione content twice that of neurons incubated in the absence of astroglial cells. In cultured neurons, the availability of cysteine limited the cellular level of glutathione. During a 4 hr incubation in a minimal medium lacking all amino acids except cysteine, the amount of neuronal glutathione was doubled. Besides cysteine, also the dipeptides CysGly and gammaGluCys were able to serve as glutathione precursors and caused a concentration-dependent increase in glutathione content. Concentrations giving half-maximal effects were 5, 5, and 200 microM for cysteine, CysGly, and gammaGluCys, respectively. In the transient cocultures, the astroglia-mediated increase in neuronal glutathione was suppressed by acivicin, an inhibitor of the astroglial ectoenzyme gamma-glutamyl transpeptidase, which generates CysGly from glutathione. These data suggest the following metabolic interaction in glutathione metabolism of brain cells: the ectoenzyme gamma-glutamyl transpeptidase uses as substrate the glutathione released by astrocytes to generate the dipeptide CysGly that is subsequently used by neurons as precursor for glutathione synthesis.

Topics: Animals; Antioxidants; Astrocytes; Cell Survival; Cells, Cultured; Coculture Techniques; Cysteine; Dipeptides; gamma-Glutamyltransferase; Glutamine; Glutathione; Glycine; Isoxazoles; Neurons; Rats; Rats, Wistar

This study was conducted to document the glutathione (GSH) cycle (interorgan circulation of GSH) in broilers in vivo. Two experiments were conducted on 36 anesthetized male broilers (n = 6 per treatment) implanted with cannulae in the carotid artery, hepatic portal, and hepatic veins. Plasma GSH, glutamate, cysteine, cystine, and cysteinylglycine levels in each vessel were monitored following a bolus injection [Experiment (Exp.) 1] or 30 min continuous infusion (Exp. 2) of GSH, or a gamma-glutamyltranspeptidase inhibitor (AT125) into the hepatic portal vein. Controls received saline alone. The GSH and AT125 treatments were used to determine the effect of increasing the prehepatic GSH load and of inhibiting systemic GSH degradation, respectively, on the GSH cycle. Hepatic export of GSH was clearly evident in all three treatment groups in both experiments (Exp.). The GSH and AT125 treatments raised amino acid levels in some or all of the vessels, whereas cysteinylglycine was elevated by AT125 and depressed by the GSH treatment compared to Controls. Hepatic uptake of glutamate, cysteine, and/or cystine was observed in Controls and GSH-treated birds, but not in birds given AT125 (Exp. 2). Neither hepatic export nor uptake of cysteinylglycine was observed in any treatment group. The results clearly demonstrate the ability of the avian liver to export GSH into the general circulation despite alterations that might arise from changes in extra-hepatic ability to utilize GSH or its constituent amino acids.

Topics: Animals; Biological Transport; Carotid Arteries; Chickens; Cysteine; Cystine; Dipeptides; Enzyme Inhibitors; gamma-Glutamyltransferase; Glutamic Acid; Glutathione; Hepatic Veins; Isoxazoles; Liver; Male; Portal Vein

The role of hepatic gamma-glutamyltransferase in the breakdown of circulating glutathione was studied in the perfused guinea pig liver. Hepatic gamma-glutamyltransferase activity in the guinea pig is sevenfold higher than in the rat and is comparable to its activity in man. Guinea pig livers were found to remove, in a single pass, 50% to 90% of glutathione (10 to 50 mumol/L) added to the portal perfusate. Removal of portal glutathione was totally dependent on the activity of gamma-glutamyltransferase and led to the near quantitative appearance of cysteinyl-glycine and cysteine in the caval perfusate. Glutathione removal by the intact liver followed saturation with a Michaelis constant (Km) of 59 mumol/L for glutathione and a maximum velocity of 235 nmol glutathione/min/gm of liver weight. The capacity of the guinea pig liver to remove circulating glutathione was estimated to be sevenfold to 10-fold higher than its net rate of output of glutathione into the circulation. Inhibition of gamma-glutamyltransferase activity in the perfused liver led to threefold to sixfold increases in the hepatic output of glutathione into the circulation, indicating that more than two thirds of glutathione transported extracellularly is broken down. Data obtained demonstrate a major role of hepatic gamma-glutamyltransferase, both in the removal of portally carried glutathione and in the degradation of glutathione molecules released by the liver itself into the sinusoids. These findings suggest the existence of an intraorgan transport of glutathione in the liver, whereby periportal cells could provide glutathione precursors to pericentral cells.

Topics: Animals; Borates; Cysteine; Dipeptides; gamma-Glutamyltransferase; Glutathione; Guinea Pigs; Isoxazoles; Liver; Perfusion; Serine