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1 Department of Physiology, Kurume University School of Medicine, Kurume 830; and 2 Laboratory of Cellular Neurobiology, Tokyo University of Pharmacy and School of Life Science, Hachioji, Tokyo 192-03, Japan
Tanaka, E., S. Yamamoto, Y. Kudo, S. Mihara, and H. Higashi. Mechanisms underlying the rapid depolarization produced by deprivation of oxygen and glucose in rat hippocampal CA1 neurons in vitro. J. Neurophysiol. 78: 891-902, 1997. Intracellular recordings were made to investigate the mechanism, site, and ionic basis of generation of the rapid depolarization induced by superfusion with ischemia-simulating medium in hippocampal CA1 pyramidal neurons of rat tissue slices. Superfusion with ischemia-simulating medium produced a rapid depolarization after ~6 min of exposure. When oxygen and glucose were reintroduced, the membrane potential did not repolarize but depolarized further, reaching 0 mV ~5 min after reintroduction. Simultaneous recordings of changes in cytoplasmic Ca2+ concentration ([Ca2+]i) and membrane potential recorded from 1-[6-amino-2-(5-carboxy-2-oxazolyl)-5-benzofuranyloxy] - 2 - ( 2 - amino - 5 - methylphenoxy ) - ethane - N, N, N
, Ntetraacetic acid pentaacetoxymethyl ester (Fura-2/AM) loaded slices revealed a rapid increase in [Ca2+]i in all CA1 layers corresponding to the rapid depolarization of the soma membrane. The result suggests that the rapid depolarization is generated not only in the soma but also in the apical and basal dendrites. Application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), DL-2-amino-4-phosphonobutyric acid, and DL-2-amino-3-phosphonopropionic acid or bicuculline did not affect the amplitude and the maximal slope. Reduction in the concentration of extracellular Ca2+ or addition of CNQX or DL-2-amino-5-phosphonopentanoic acid delayed the onset of the rapid depolarization. The amplitude of the rapid depolarization recorded with Cs acetate electrodes in tetraethylammonium-containing medium had a linear relationship to the membrane potential between 
50 and 20 mV. The reversal potential was shifted in the hyperpolarizing direction by a decrease in either [Na+]o or [Ca2+]o, whereas the reversal potential was shifted in the depolarizing direction by a decrease in [Cl
]o or using CsCl electrodes. An increase or decrease in [K+]o did not affect the reversal potential. These results indicate that the rapid depolarization is Na+, Ca2+, and Cl dependent. The lack of effects of changes in [K+]o is probably due to the accumulation of interstitial K+ before generating the rapid depolarization. Prolonged application of ouabain (30 µM) caused an initial small hyperpolarization, a subsequent slow depolarization, and a rapid depolarization. In summary, the present study has demonstrated that the rapid depolarization is voltage-independent and is probably due to a nonselective increase in permeability to all participating ions, which may occur only in pathological conditions. The underlying conductance change is primarily the result of inhibition of Na,K-ATPase activity in the recorded neuron.
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