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Journal of Neuroscience, Vol 9, 1897-1909, Copyright © 1989 by Society for Neuroscience
Dynamics of the endoplasmic reticulum and other membranous organelles in growth cones of cultured neurons
ME Dailey and PC Bridgman
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110.
The fluorescent lipophilic dye 3,3'-dihexyloxacarbocyanine iodide
[DiOC6(3)] was used to examine the distribution of membrane-bound
organelles in growth cones of cultured rat sympathetic neurons. Within
chemically fixed growth cones, intense DiOC6(3) fluorescence was localized
predominately to the base or central region of growth cones. However, in
most growth cones several thin DiOC6(3)-fluorescent processes radiated from
the base into the periphery, and double fluorescence imaging of single
growth cones indicated that these processes were highly colocalized
(approximately 79%) with microtubules. The distribution of DiOC6(3)
fluorescence in living growth cones was examined using low light-level
fluorescence video microscopy. We observed thin fluorescent processes
within the periphery of growth cones to undergo length excursions
(extension/retraction) and to change orientation (move laterally). During
growth cone advance, processes became progressively thicker and were
gradually engulfed by the advancing fluorescent mass. When growth cones
were viewed with video-enhanced differential interference contrast
microscopy, the position of the fluorescent processes correlated with
thickened extensions of central-type cytoplasm through which vesiclelike
organelle transport often occurred. These observations indicate several
features concerning the organization and movement of membranous organelles
(MOs) in growth cones: (1) MOs are highly compartmentalized, the majority
being localized to the growth cone base; (2) MOs advance into the periphery
along distinct pathways probably associated with microtubules; (3) one or
more thin continuous MOs, which most likely represent a thin tubular
component of the endoplasmic reticulum, generally precedes advance of
vesiclelike MOs along individual transport pathways; and (4) transport
pathways with their associated MOs are spatially and temporally dynamic.
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