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The Journal of Neuroscience, April 1, 2001, 21(7):2393-2403
Dendritic Spines Lost during Glutamate Receptor Activation
Reemerge at Original Sites of Synaptic Contact
M. Josh
Hasbani,
Michelle L.
Schlief,
Daniel A.
Fisher, and
Mark P.
Goldberg
Departments of Neurology and Anatomy and Neurobiology Center for
the Study of Nervous System Injury, Washington University School of
Medicine, St. Louis, Missouri 63110
During cerebral ischemia, neurons undergo rapid alterations in
dendritic structure consisting of focal swelling and spine loss. We
used time-lapse microscopy to determine the fate of dendritic spines
that disappeared after brief, sublethal hypoxic or excitotoxic exposures. Dendrite and spine morphology were assessed in cultured cortical neurons expressing yellow fluorescent protein or labeled with
the fluorescent membrane tracer, DiI. Neurons exposed to NMDA, kainate,
or oxygen-glucose deprivation underwent segmental dendritic beading
and loss of approximately one-half of dendritic spines. Most spine loss
was observed in regions of local dendritic swelling. Despite widespread
loss, spines recovered within 2 hr after termination of agonist
exposure or oxygen-glucose deprivation and remained stable over the
subsequent 24 hr. Recovery was slower after NMDA than AMPA/kainate
receptor activation. Time-lapse fluorescence imaging showed that the
vast majority of spines reemerged in the same location from which they
disappeared. In addition to spine recovery, elaboration of dendritic
filopodia was observed in new locations along the dendritic shaft after
dendrite recovery. Spine recovery did not depend on actin
polymerization because it was not blocked by application of
latrunculin-A, which eliminated filamentous actin staining in spines
and blocked spine motility. Throughout spine loss and recovery,
presynaptic and postsynaptic elements remained in physical proximity.
These results suggest that elimination of dendritic spines is not
necessarily associated with loss of synaptic contacts. Rapid
reestablishment of dendritic spine synapses in surviving neurons may be
a substrate for functional recovery after transient cerebral ischemia.
Key words:
hypoxia; glutamate; excitotoxicity; dendritic spine; synapse; actin
Copyright © 2001 Society for Neuroscience 0270-6474/01/2172393-11$05.00/0
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