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Journal of Neuroscience, Vol 13, 1993-2000, Copyright © 1993 by Society for Neuroscience
Glutamate receptor-induced 45Ca2+ accumulation in cortical cell culture correlates with subsequent neuronal degeneration
DM Hartley, MC Kurth, L Bjerkness, JH Weiss and DW Choi
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri 63110.
Murine neuronal and glial cell cultures exposed briefly to glutamate
accumulated large amounts of 45Ca2+ from the extracellular medium during
the exposure. Most of the accumulation likely reflected influx into
neurons, as little accumulation was observed in similarly treated glial
cultures. When the concentration of glutamate was varied between 10 and
1000 microM, or exposure duration was varied between 0 and 10 min, the
amount of 45Ca2+ accumulation correlated closely with the amount of
neuronal death 24 hr later. Both 45Ca2+ accumulation and cell death could
be attenuated in a dose-dependent manner by the competitive NMDA antagonist
D-aminophosphonovalerate or the noncompetitive antagonist dextrorphan, with
IC50 values of approximately 100 microM and 15 microM, respectively. In
contrast, neither 45Ca2+ accumulation nor cell death was blocked by the
alpha-amino-3-hydroxy-5-methyl-4- isoxazole propionic acid (AMPA)/kainate
antagonist 6-cyano-7- nitroquinoxaline-2,3-dione (CNQX) in the presence of
high glycine. With brief exposure, high concentrations of AMPA, kainate, or
K+ produced much less death or 45Ca2+ accumulation than produced by
glutamate, especially if 10 microM MK-801 was included in the exposure
medium to block NMDA receptor activation. Kainate- or AMPA-induced 45Ca2+
accumulation or neuronal cell death was blocked with CNQX. However, high
K(+)-triggered 45Ca2+ accumulation was only partially blocked with CNQX
plus MK-801, consistent with mediation by voltage-gated Ca2+ channels. In
addition to measuring the accumulation of 45Ca2+ occurring during agonist
exposure, we also assessed accumulation during the 30 min immediately
following completion of a 3-5 min exposure to 500 microM NMDA.(ABSTRACT
TRUNCATED AT 250 WORDS)
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