Single-molecule discrimination of discrete perisynaptic and distributed sites of actin filament assembly within dendritic spines.
Within dendritic spines, actin is presumed to anchor receptors in the postsynaptic density and play numerous roles regulating synaptic transmission. However, the submicron dimensions of spines have hindered examination of actin dynamics within them and prevented live-cell discrimination of perisynaptic actin filaments. Using photoactivated localization microscopy, we measured movement of individual actin molecules within living spines. Velocity of single actin molecules along filaments, an index of filament polymerization rate, was highly heterogeneous within individual spines. Most strikingly, molecular velocity was elevated in discrete, well-separated foci occurring not principally at the spine tip, but in subdomains throughout the spine, including the neck. Whereas actin velocity on filaments at the synapse was substantially elevated, at the endocytic zone there was no enhanced polymerization activity. We conclude that actin subserves spatially diverse, independently regulated processes throughout spines. Perisynaptic actin forms a uniquely dynamic structure well suited for direct, active regulation of the synapse.
Frost
Nicholas A
NA
Department of Physiology and Program in Neuroscience, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Shroff
Hari
H
Kong
Huihui
H
Betzig
Eric
E
Blanpied
Thomas A
TA
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D049229:Dendritic Spines / Q000378:metabolism*; D004622:Embryo, Mammalian; D049452:Green Fluorescent Proteins / Q000235:genetics; D006624:Hippocampus / Q000166:cytology; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009473:Neuronal Plasticity / Q000502:physiology; D009474:Neurons / Q000166:cytology*; D017729:Presynaptic Terminals / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley; D009435:Synaptic Transmission / Q000502:physiology; D014018:Tissue Distribution; D014162:Transfection / Q000379:methods
D009419:Nerve Tissue Proteins; D049452:Green Fluorescent Proteins