The lack of extracellular Na+ exacerbates Ca2+-dependent damage of cultured cerebellar granule cells.
Rhodamine 123 staining, light and electron microscopy were used to evaluate the ultrastructural and functional state of cultured cerebellar granule cells after short treatment with the solution where NaCl was substituted by sucrose (sucrose balance salt medium, SBSM). Cell exposure to SBSM for 20 min resulted in the fact that mitochondria in the neurons lost their ability to sequester rhodamine 123. This effect could be prevented by: (i) non-competitive N-methyl-D-aspartate (NMDA) receptor channel blocker, 10(-5) M MK-801; (ii) a competitive specific antagonist of NMDA glutamate receptors, 0.25 x 10(-3) M D,L-2-amino-7-phosphonoheptanoate (APH); (iii) 10(-3) M cobalt chloride; (iv) removal of Ca2+ from the medium. Low Na+ in the Ca2+-containing medium caused considerable mitochondrial swelling in granule cells. However, the same treatment in the absence of calcium ions in the medium abolished the deleterious effect of SBSM on the neuronal mitochondrial structure and functions. It is suggested that (i) the exposure of cultured cerebellar granule cells to SBSM leads to a release of endogenous glutamate from cells; (ii) Ca2+ ions potentially de-energizing neuronal mitochondria enter the neuron preferentially through the NMDA channels rather than through the Na+/Ca2+ exchanger; (iii) mitochondrial swelling in granule cells is highly Ca2+-dependent; (iv) cellular overload with sodium ions can activate mitochondrial Na+/Ca2+ exchanger and thus prevent permeability transition pore opening in mitochondria.
Isaev
N K
NK
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia. isaev@energ.genebee.msu.su
Stelmashook
E V
EV
Alexandrova
O P
OP
Andreeva
N A
NA
Polyakova
I A
IA
Victorov
I V
IV
Zorov
D B
DB
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D002531:Cerebellum / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D003035:Cobalt / Q000494:pharmacology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005456:Fluorescent Dyes; D008854:Microscopy, Electron; D051381:Rats; D017208:Rats, Wistar; D020112:Rhodamine 123; D012235:Rhodamines; D012965:Sodium Chloride / Q000633:toxicity*
D018691:Excitatory Amino Acid Antagonists; D005456:Fluorescent Dyes; D012235:Rhodamines; D020112:Rhodamine 123; D003035:Cobalt; D012965:Sodium Chloride; D016291:Dizocilpine Maleate; C018021:cobaltous chloride; D002118:Calcium