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Journal of Neuroscience, Vol 4, 1944-1953, Copyright © 1984 by Society for Neuroscience
An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. I. Cells which develop without intercellular contacts
WP Bartlett and GA Banker
We have studied the processes which are elaborated by hippocampal neurons
in dissociated cell culture. Nerve cells, which were obtained from fetal
rats at 18 to 20 days of gestation, were plated at very low density onto
polylysine-treated coverslips and were maintained in serum- free medium.
Under such conditions, some cells develop without contacting any
neighboring neurons or glial cells. Examples of such isolated cells which
had developed for 1 week in culture were studied first by light microscopy,
then they were sectioned parallel to the substratum so that all portions of
the cell and its processes could be examined by electron microscopy.
Dendrites and axons could be clearly distinguished by both light and
electron microscopy. Dendrites were rather thick at the base but tapered
rapidly to a minimum diameter of about 0.5 micron and contained
polyribosomes throughout their length. Axons, which were several times
longer than the dendrites, were thinner at the origin, tapered much less,
and were essentially ribosome-free. These ultrastructural differences were
particularly obvious at branch points, where cytoplasmic organelles tend to
accumulate. Clusters of polyribosomes were invariably present at dendritic
branch points, but they were never observed at axonal branch points. The
axons most commonly arose from the proximal portion of a dendrite rather
than directly from the cell body as they typically do in situ. These
observations show that the fundamental differences in form and in the
distribution of ribosomes between axons and dendrites can be established in
cell culture. Contact with afferent fibers or with target cells during the
period of process outgrowth is unnecessary for the expression of these
features of axonal and dendritic differentiation.
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