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Journal of Neuroscience, Vol 14, 2585-2593, Copyright © 1994 by Society for Neuroscience
Perturbation of intracellular calcium and hydrogen ion regulation in cultured mouse hippocampal neurons by reduction of the sodium ion concentration gradient
RA Koch and ME Barish
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010.
Na(+)-Ca2+ exchange has been identified as a mechanism for regulation of
intracellular Ca ion concentration ([Ca2+]i) in neurons of invertebrates
and vertebrates, but for mammalian central neurons its role in restoration
of resting [Ca2+]i after transient increases induced by stimulation has
been less clear. We have examined the recovery of [Ca2+]i following K+
depolarization and glutamate receptor activation of cultured mouse
hippocampal neurons using the Ca(2+)- sensitive dye Fura-2. Reduction of
the transmembrane Na+ gradient by removal of external Na+ slowed the
recovery of neurons from imposed Ca2+ loads. We observed that [Ca2+]i
regulation was disrupted more severely when N-methyl-D-glucamine (N-MG),
Tris, or choline rather than Li+ replaced external Na+. Additional
disruption of intracellular pH regulation by substitutes other than Li+ may
account for this difference. Measurement of [Ca2+]i and [H+]i (using the
H(+)-sensitive dye BCECF) during glutamate receptor activation indicated
that Ca2+ influx resulted in production of intracellular H+, and that Li+
but not N-MG could prevent cytoplasmic acidification on removal of external
Na+. We also observed that intracellular acidification alone was sufficient
to slow recovery from Ca2+ load. We conclude, therefore, that Na(+)-Ca2+
exchange contributes to recovery of [Ca2+]i after stimulation leading to
Ca2+ entry into hippocampal neurons, and that Na(+)-H+ exchange limits the
acidification (and secondary increase in [Ca2+]i) that accompanies Ca2+
influx. We suggest that because both Na(+)-Ca2+ and Na(+)-H+ exchangers
will be compromised during ischemia and hypoglycemia, increased
intracellular H+ may synergize with cytoplasmic Ca2+ to potentiate
excitotoxic neuronal death.
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