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Journal of Neuroscience, Vol 8, 1425-1435, Copyright © 1988 by Society for Neuroscience
Growth cone configuration and advance: a time-lapse study using video- enhanced differential interference contrast microscopy
JM Aletta and LA Greene
Department of Pharmacology, NYU Medical Center, New York 10016.
We have analyzed the dynamics of growth cone configuration using video-
enhanced contrast differential interference contrast microscopy.
Regenerating neurites from NGF-treated PC12 cells and sympathetic neurons
were observed in real time during their elongation and reviewed by
time-lapse video recordings. This technique provided a high- resolution
view of motile growth cone elements including filopodia, microspikes,
lamellipodia and ruffles. On the basis of our observations, a multistage
model for growth cone advance is proposed. Elongation commences with
lamellipodial spreading. If the newly extended lamellipodium does not
retract or lift off the substrate in the form of a ruffle, a second
phase--consolidation--occurs, in which the lamellipodium thickens as it
fills with cytoplasm and organelles. The consolidated area then undergoes
further transformation into an area of neuritic shaft as new lamellipodia
form at the leading and peripheral zones of the distal process. We never
observed filopodia or microspikes contracting to propel the growth cone
forward. We also noted that elongating tips generally had large
varicosities within 20 micron of their leading edges. These may play a role
in neurite outgrowth and in the formation of smaller, synaptic
vesicle-containing varicosities. The dynamic behavior of the growth cone
was under the control of NGF. Withdrawal of NGF resulted in the
disappearance of motile structures and cessation of growth, while
readdition of NGF triggered the rapid reappearance of these structures and
the resumption of growth. The high-resolution video microscopy of living
growth cones provides necessary baseline information, as well as a bioassay
paradigm, for future studies on the molecular mechanisms of nerve growth.
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