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APPETITE, OBESITY, DIGESTION, AND METABOLISM

Na-K-ATPase in rat cerebellar granule cells is redox sensitive

¹Institute of Veterinary Physiology, Vetsuisse Faculty and Zurich Centre of Integrative Human Physiology, University of Zurich, Zurich, Switzerland; ²Department of Biochemistry, International Biotechnological Center of Lomonosov Moscow State University, Moscow, Russia; and ³Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

Submitted 19 January 2005 ; accepted in final form 8 November 2005

Redox-induced regulation of the Na-K-ATPase was studied in dispersed rat cerebellar granule cells. Intracellular thiol redox state was modulated using glutathione (GSH)-conjugating agents and membrane-permeable ethyl ester of GSH (et-GSH) and Na-K-ATPase transport and hydrolytic activity monitored as a function of intracellular reduced thiol concentration. Depletion of cytosolic and mitochondrial GSH pools caused an increase in free radical production in mitochondria and rapid ATP deprivation with a subsequent decrease in transport but not hydrolytic activity of the Na-K-ATPase. Selective conjugation of cytosolic GSH did not affect free radical production and Na-K-ATPase function. Unexpectedly, overloading of cerebellar granule cells with GSH triggered global free radical burst originating most probably from GSH autooxidation. The latter was not followed by ATP depletion but resulted in suppression of active K⁺ influx and a modest increase in mortality. Suppression of transport activity of the Na-K-ATPase was observed in granule cells exposed to both permeable et-GSH and impermeable GSH, with inhibitory effects of external and cytosolic GSH being additive. The obtained data indicate that redox state is a potent regulator of the Na-K-ATPase function. Shifts from an "optimal redox potential range" to higher or lower levels cause suppression of the Na-K pump activity.

glutathione; free radicals; redox state

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