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Volume 16, Number 13,
Issue of July 1, 1996
pp. 4069-4079
Copyright ©1996 Society for Neuroscience
Motor Neurons Are Selectively Vulnerable to AMPA/Kainate
Receptor-Mediated Injury In Vitro
Received Feb. 12, 1996; revised April 1, 1996; accepted April 8, 1996.
Sean G. Carriedo1,
Hong Z. Yin2, and
John H. Weiss1, 2, 3
Departments of 1 Psychobiology,
2 Neurology, and 3 Anatomy and Neurobiology,
University of California-Irvine, Irvine, California 92717-4290
The nonphosphorylated neurofilament marker SMI-32 stains motor
neurons in spinal cord slices and stains a subset of cultured spinal
neurons [``large SMI-32(+) neurons''], which have a morphology
consistent with motor neurons identified in vitro: large
cell body, long axon, and extensive dendritic arborization. They are
found preferentially in ventral spinal cord cultures, providing further
evidence that large SMI-32(+) neurons are indeed motor neurons, and
SMI-32 staining often colocalizes with established motor neuron markers
(including acetylcholine, calcitonin gene-related peptide, and
peripherin). Additionally, choline acetyltransferase activity (a
frequently used index of the motor neuron population) and peripherin(+)
neurons share with large SMI-32(+) neurons an unusual vulnerability to
AMPA/kainate receptor-mediated injury. Kainate-induced loss of
these motor neuron markers is Ca2+-dependent,
which supports a critical role of Ca2+ ions in
this injury. Raising extracellular Ca2+
exacerbates injury, whereas removal of extracellular
Ca2+ is protective. A basis for this
vulnerability is provided by the observation that most peripherin(+)
neurons, like large SMI-32(+) neurons, are subject to
kainate-stimulated Co2+ uptake, a histochemical
stain that identifies neurons possessing
Ca2+-permeable AMPA/kainate
receptor-gated channels. Finally, of possibly greater relevance to the
slow motor neuronal degeneration in diseases, both large SMI-32(+)
neurons and peripherin(+) neurons are selectively damaged by prolonged
(24 hr) low-level exposures to kainate (10 µM)
or to the glutamate reuptake blocker
L-trans-pyrrolidine-2,4-dicarboxylic
acid (100 µM). During these low-level kainate
exposures, large SMI-32(+) neurons showed higher intracellular
Ca2+ concentrations than most spinal neurons,
suggesting that Ca2+ ions are also important in
this more slowly evolving injury.
Key words:
cell culture;
AMPA;
kainate;
glutamate;
motor neuron;
calcium;
SMI-32;
peripherin;
ChAT;
neurotoxicity;
cobalt;
calcium
imaging
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