Modulation of glutamate exocytosis by redox changes of superficial thiol groups in rat cerebrocortical synaptosomes.
The treatment of cerebral cortex synaptosomes with the membrane impermeable thiol reagent 5,5'-dithio-bis-(2-nitrobenzoic acid) (DTNB) induces a long-lasting partial inhibition (about 40%) of the KCl-stimulated Ca2+-dependent exocytosis of glutamate. Synaptosomes are not damaged by the treatment. The increase of cytoplasmic free Ca2+ concentration ([Ca2+]i) upon depolarization is not affected by DTNB. The inhibition is observed also if exocytosis is induced with the Ca2+-ionophore ionomycin. In all cases the inhibition is reversed by the impermeable reductant glutathione (GSH). Similarly the inhibition of exocytosis by H2O2 (Zoccarato, F., Valente, M. and Alexandre, A., Hydrogen peroxide induces a long-lasting inhibition of the Ca2+-dependent glutamate release in cerebrocortical synaptosomes without interfering with cytosolic Ca2+. J. Neurochem., 64 (1995) 2552-2558.) is reversed by GSH. It is concluded that redox changes (possibly thiol-disulfide transitions) of superficial groups modulate the exocytotic apparatus directly. In an attempt to identify the protein(s) involved in this novel type of control, we evidenced DTNB (H2O2) reactive bands at 35 and at 85-150 kDa which can be labeled with a monobromotrimethylammoniobimane bromide (qBBr) derivatization.
Zoccarato
F
F
Department of Biological Chemistry and C.N.R. Unit for the Study of Biomembranes, University of Padova, Italy.
Cavallini
L
L
Valente
M
M
Alexandre
A
A
D000818:Animals; D002118:Calcium / Q000493:pharmacokinetics; D002540:Cerebral Cortex / Q000378:metabolism; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004586:Electrophoresis; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D005456:Fluorescent Dyes; D018698:Glutamic Acid / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D000644:Quaternary Ammonium Compounds; D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism*
D005456:Fluorescent Dyes; D016877:Oxidants; D000644:Quaternary Ammonium Compounds; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D018698:Glutamic Acid; C027344:bromotrimethylammoniobimane; D004228:Dithionitrobenzoic Acid; D006861:Hydrogen Peroxide; D005978:Glutathione; D002118:Calcium