13C NMR spectroscopic measurement of glutathione synthesis and antioxidant metabolism in the intact ocular lens.
A 13C NMR spectroscopic method for non-invasive, time-resolved measurements of glutathione function in the intact ocular lens maintained in organ culture is described. L-[beta-13C]cysteine (1 mM) included in the incubation medium is incorporated, by way of lenticular amino acid uptake and glutathione biosynthetic mechanisms, into the cysteinyl residue of intralenticular glutathione. 13C-NMR chemical shift measurements facilitate analysis of glutathione synthesis and anti-oxidant reactions in the intact tissue. The results of this preliminary study demonstrate the viability of a rapid non-invasive method for monitoring the multiple aspects of glutathione biosynthesis, metabolism, and function in intact tissue.
Willis
J A
JA
Department of Chemistry and Biochemistry, University of California, Santa Cruz 95064.
Schleich
T
T
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D002247:Carbon Isotopes; D003545:Cysteine / Q000378:metabolism*; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism*; D019803:Glutathione Disulfide; D007700:Kinetics; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009924:Organ Culture Techniques; D016877:Oxidants / Q000494:pharmacology; D010545:Peroxides / Q000494:pharmacology; D011817:Rabbits; D013997:Time Factors; D020122:tert-Butylhydroperoxide
D000975:Antioxidants; D002247:Carbon Isotopes; D016877:Oxidants; D010545:Peroxides; D020122:tert-Butylhydroperoxide; D005978:Glutathione; D003545:Cysteine; D019803:Glutathione Disulfide