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Journal of Neuroscience, Vol 14, 1130-1140, Copyright © 1994 by Society for Neuroscience
Development of subcellular mRNA compartmentation in hippocampal neurons in culture
R Kleiman, G Banker and O Steward
Department of Neuroscience, University of Virginia Health Sciences Center, Charlottesville 22908.
Neurons possess an RNA transport system that is present in dendrites (but
not axons) and sort mRNAs so that some mRNAs are restricted to cell bodies
while a few others (like the mRNA for MAP2) are present in dendrites. The
present study evaluates when dendrite-specific RNA transport and mRNA
sorting into cell body and somatodendritic compartments first appear in
developing hippocampal neurons maintained in culture. A 3H-uridine
pulse-chase paradigm was used to evaluate transport of newly synthesized
RNA from the site of synthesis in the nucleus into the developing neurites.
The intracellular distribution of mRNAs encoding actin, tubulin, GAP-43,
and MAP2 as well as polyA RNA and rRNA was evaluated by in situ
hybridization at different stages of development. Newly synthesized RNA was
translocated into both developing axons and dendrites early in development,
but only into dendrites as the neurons matured. Tubulin, GAP-43, and actin
mRNAs, which are restricted to cell bodies in mature neurons, were found
exclusively in neuronal cell bodies at all developmental stages. MAP2 mRNA,
which is present in the dendrites of mature neurons, was present at very
low levels in neurons at 2 or 3 d in culture and was not detectable within
dendrites. The overall levels of MAP2 mRNA increased over time, and by 5-7
d in culture, MAP2 mRNA was detectable in some dendrites. PolyA RNA and
rRNA were detectable in developing neurites including axons. Levels of
polyA and rRNA increased in dendrites as neurons matured while labeling of
axons diminished. By 10 d in culture, axonal labeling for polyA and rRNA
had virtually disappeared. The increase in the levels of polyA, rRNA, and
MAP2 mRNA in dendrites between 5 and 7 d in culture corresponds roughly
with the appearance of other dendritic characteristics and the beginning of
dendritic outgrowth.
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