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Journal of Neuroscience, Vol 11, 1918-1929, Copyright © 1991 by Society for Neuroscience
Actin dynamics in growth cones
S Okabe and N Hirokawa
Department of Anatomy and Cell Biology, School of Medicine, University of Tokyo, Japan.
The mechanism of actin incorporation and turnover in the nerve growth cone
was examined by immunoelectron microscopy and low-light-level video
microscopy of cultured neurons injected with biotin-labeled actin or
fluorescently labeled actin. We first determined the sites of actin
incorporation into the cytoskeleton of growth cones by immunoelectron
microscopy of cultured neurons injected with biotin-labeled actin and
reacted with an anti-biotin antibody and a gold-labeled secondary antibody.
Shortly after the injection, biotin-actin molecules incorporated into the
cytoskeleton were localized in the distal part of actin bundles in the
filopodia and at the membrane-associated fringe of the actin filament
network. With longer incubation, most actin polymers in the growth cones
were labeled uniformly, suggesting that actin subunits are added
preferentially at the membrane-associated ends of preexisting actin
filaments. We then determined whether actin filaments translocate within
the growth cones by low-light-level video microscopy of living neurons
injected with fluorescently labeled actin and photobleached with a laser
beam. When actin fluorescence at the leading edge of a growth cone was
bleached, a rearward translocation of the bleached spot toward the base of
the growth cone was observed. This observation suggests the presence of a
rearward flow of actin polymers within growth cones. Taken together, these
results indicate that there is a continuous addition of actin monomers at
the leading edge of the growth cone and a successive rearward translocation
of the assembled filaments.
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