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The Journal of Neuroscience, January 15, 1999, 19(2):619-629
Oxygen/Glucose Deprivation in Hippocampal Slices: Altered
Intraneuronal Elemental Composition Predicts Structural and Functional
Damage
Charles P.
Taylor1,
Mark L.
Weber1,
Christopher
L.
Gaughan2,
Ellen J.
Lehning2, and
Richard M.
LoPachin2
1 Department of Neuroscience Therapeutics, Parke-Davis
Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor,
Michigan 48105, and 2 Department of Anesthesiology, Albert
Einstein College of Medicine, Montefiore Medical Center, Bronx, New
York 10467
Effects of oxygen/glucose deprivation (OGD) on subcellular
elemental composition and water content were determined in nerve cell
bodies from CA1 areas of rat hippocampal slices. Electron probe x-ray
microanalysis was used to measure percentage water and concentrations
of Na, P, K, Cl, Mg, and Ca in cytoplasm, nucleus, and mitochondria of
cells exposed to normal and oxygen/glucose deficient medium. As an
early (2 min) consequence of OGD, evoked synaptic potentials were lost,
and K, Cl, P, and Mg concentrations decreased significantly in all
morphological compartments. As exposure to in vitro OGD
continued, a negative DC shift in interstitial voltage occurred (~5
min), whereas general elemental disruption worsened in cytoplasm
and nucleus (5-42 min). Similar elemental changes were noted in
mitochondria, except that Ca levels increased during the first 5 min of
OGD and then decreased over the remaining experimental period (12-42
min). Compartmental water content decreased early (2 min), returned to
control after 12 min of OGD, and then exceeded control levels at 42 min. After OGD (12 min), perfusion of hippocampal slices with control
oxygenated solutions (reoxygenation) for 30 min did not restore
synaptic function or improve disrupted elemental composition. Notably,
reoxygenated CA1 cell compartments exhibited significantly elevated Ca
levels relative to those associated with 42 min of OGD. When slices
were incubated at 31°C (hypothermia) during OGD/reoxygenation,
neuronal dysfunction and elemental deregulation were minimal. Results
show that in vitro OGD causes loss of transmembrane Na,
K, and Ca gradients in CA1 neurons of hippocampal slices and that
hypothermia can obtund this damaging process and preserve neuronal function.
Key words:
oxygen/glucose deprivation; ischemia; reperfusion injury; CA1 cells; hypothermia; neuroprotection; electron probe x-ray
microanalysis
Copyright © 1999 Society for Neuroscience 0270-6474/99/192619-11$05.00/0
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