Lippincott Williams & Wilkins, Inc., Philadelphia
Glutamate Neurotoxicity in Rat Cerebellar Granule Cells Involves Cytochrome c Release from Mitochondria and Mitochondrial Shuttle Impairment
Article first published online: 18 JAN 2002
Journal of Neurochemistry
Volume 73, Issue 1, pages 237–246, July 1999
How to Cite
Atlante, A., Gagliardi, S., Marra, E., Calissano, P. and Passarella, S. (1999), Glutamate Neurotoxicity in Rat Cerebellar Granule Cells Involves Cytochrome c Release from Mitochondria and Mitochondrial Shuttle Impairment. Journal of Neurochemistry, 73: 237–246. doi: 10.1046/j.1471-4159.1999.0730237.x
Abbreviations used: ADK, adenylate kinase; BME, basal medium with Eagle’s salts; CGCs, cerebellar granule cells; cyt c, cytochrome c; DHAP, dihydroxyacetone phosphate; e.u., enzymatic units; GDH, glutamate dehydrogenase; Glu-CGCs, glutamate-treated CGCs; GNT, glutamate neurotoxicity; G3P, glycerol 3-phosphate; G3PDH, glycerol-3-phosphate dehydrogenase; MAL, malate; MDH, malate dehydrogenase; MK-801, (5R, 10S)-(+)-5-methyl-10,11-dihydro[a,d]cyclohepten-5,10-imine hydrogen maleate; OAA, oxaloacetate; PBS, phosphate-buffered saline; TMPD, N,N,N′,N′-tetramethyl-p-phenylenediamine.
- Issue published online: 18 JAN 2002
- Article first published online: 18 JAN 2002
- Cytochrome c;
Abstract: To gain some insight into the mechanism by which glutamate neurotoxicity takes place in cerebellar granule cells, two steps of glucose oxidation were investigated: the electron flow via respiratory chain from certain substrates to oxygen and the transfer of extramitochondrial reducing equivalents via the mitochondrial shuttles. However, cytochrome c release from intact mitochondria was found to occur in glutamate-treated cells as detected photometrically in the supernatant of the cell homogenate suspension. As a result of cytochrome c release, an increase of the oxidation of externally added NADH was found, probably occurring via the NADH-b5 oxidoreductase of the outer mitochondrial membrane. When the two mitochondrial shuttles glycerol 3-phosphate/dihydroxyacetone phosphate and malate/oxaloacetate, devoted to oxidizing externally added NADH, were reconstructed, both were found to be impaired under glutamate neurotoxicity. Consistent early activation in two NADH oxidizing mechanisms, i.e., lactate production and plasma membrane NADH oxidoreductase activity, was found in glutamate-treated cells. In spite of this, the increase in the cell NADH fluorescence was found to be time-dependent, an index of the progressive damage of the cell.