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Journal of Neuroscience, Vol 16, 1346-1358, Copyright © 1996 by Society for Neuroscience
Localization and active transport of mRNA in axons of sympathetic neurons in culture
M Olink-Coux and PJ Hollenbeck
Department of Neurobiology, Harvard Medical School, Boston, Massachusetts 02115, USA.
In neurons, the establishment and maintenance of distinct somatic,
dendritic, and axonal domains has long been known to rely on regulated
traffic of organelles and proteins. More recently, the local targeting of
specific mRNAs has also been demonstrated, at least in dendrites, to
provide a local supply of specific proteins. Here we set out to test
directly for the presence of mRNA in axons of cultured chick sympathetic
neurons, to examine their distribution during axonal outgrowth, to
determine the reliance of this distribution on specific cytoskeletal
elements, and to assess whether the axonal and somatic mRNA complements
differ. Using fluorescent in situ hybridization, we found that sympathetic
axons do contain poly(A+) mRNA along their length in a pattern that changes
gradually as axons elongate, from an evenly dispersed punctate distribution
with strong growth cone staining to a distribution focused at branch
points, varicosities, and some growth cones. Selective perturbations of the
cytoskeleton revealed that the presence of axonal mRNA was dependent on
microtubules (MTs), but not actin filaments, indicating that mRNA transport
and/or anchoring within the axon are active processes involving MTs.
Finally, reverse transcription-PCR amplification of RNAs from the axonal
and somatic compartments showed that beta-actin mRNA was present in both
compartments, whereas mRNA encoding alpha-tubulin was restricted to the
somatic compartment and entirely absent from the axons. Thus, the mRNA
populations in the soma versus the axon are both quantitatively and
qualitatively different, and these neurons are able to direct specific
mRNAs to the axon.
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