Regulation of F-actin stability in dendritic spines by glutamate receptors and calcineurin.
Neuronal degeneration and cell death can result from excessive activation of receptors for the excitatory neurotransmitter glutamate; however, the very earliest changes in cytoskeletal organization have not been well documented. We have used an in vitro model system to examine the early consequences of intense glutamate receptor activation on dendritic spine synapses. Cultured hippocampal neurons exposed to NMDA for as little as 5 min exhibited a rapid and extensive loss of dendritic spines. Staining for the presynaptic marker synapsin 1 and the postsynaptic density proteins PSD-95 and the NR1 subunit of NMDA receptors remained intact. The disappearance of spines was accompanied by a selective loss of filamentous actin staining at synapses. The NMDA-induced loss of spine actin was time- and concentration-dependent and blocked by NMDA receptor antagonists. The effect was mimicked by L-glutamate, AMPA, and ionomycin but not by agonists of L-type calcium channels or of metabotropic glutamate receptors. The effect of NMDA on local actin assembly was strongly attenuated by pretreatment with an actin stabilizing compound or by an antagonist of the calcium-dependent protein phosphatase calcineurin. Immunoreactivity for calcineurin was enriched at synapses together with F-actin. These results indicate that the actin-mediated stability of synaptic structure is disrupted by intense glutamate receptor activity and that calcineurin blockers may be useful in preventing such destabilization.
Halpain
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S
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Hipolito
A
A
Saffer
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L
D000199:Actins / Q000378:metabolism*; D000818:Animals; D019703:Calcineurin / Q000378:metabolism*; D048429:Cell Size / Q000187:drug effects; D003599:Cytoskeleton / Q000378:metabolism; D003712:Dendrites / Q000737:chemistry / Q000378:metabolism*; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology; D016202:N-Methylaspartate / Q000494:pharmacology; D017729:Presynaptic Terminals / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D017470:Receptors, Glutamate / Q000378:metabolism*
D000199:Actins; D018690:Excitatory Amino Acid Agonists; D017470:Receptors, Glutamate; D016202:N-Methylaspartate; D019703:Calcineurin