Screening of thiol compounds: depolarization-induced release of glutathione and cysteine from rat brain slices.
Superfusates from rat brain slices were screened for thiol compounds after derivatization with monobromobimane by reversed-phase HPLC. Only glutathione and cysteine were detected. The Ca(2+)-dependent release of these compounds from slices of different regions of rat brain was investigated, applying a highly sensitive and reproducible quantification method, based on reduction of superfusates with dithiothreitol, reaction of thiols with iodoacetic acid, precolumn derivatization with o-phthalaldehyde reagent solution, and analysis with reversed-phase HPLC. This methodology allowed determination of reduced and total thiols in aliquots of the same superfusates. Mostly reduced glutathione and cysteine were released upon K+ depolarization and the Ca2+ dependency suggests that they originate from a neuronal compartment. The GSH release was most prominent in the mesodiencephalon, cortex, hippocampus, and striatum and lowest in the pons-medulla and cerebellum. This underscores a physiologically significant role for glutathione in CNS neurotransmission.
Brain Research Institute, University of Zürich, Switzerland.
L
L
Zängerle
M
M
Cuénod
K H
KH
Winterhalter
K Q
KQ
Do
D000818:Animals; D001377:Azaserine / Q000494:pharmacology; D001921:Brain / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000378:metabolism*; D003980:Diazooxonorleucine / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism*; D010084:Oxidation-Reduction; D010477:Perfusion; D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D005723:gamma-Glutamyltransferase / Q000037:antagonists & inhibitors
D013438:Sulfhydryl Compounds; D003980:Diazooxonorleucine; D001377:Azaserine; D005723:gamma-Glutamyltransferase; D005978:Glutathione; D003545:Cysteine