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The Journal of Neuroscience, January 1, 1998, 18(1):251-265
Sorting of -Actin mRNA and Protein to Neurites and Growth
Cones in Culture
Gary J.
Bassell1,
Honglai
Zhang1,
Anne L.
Byrd1,
Andrea M.
Femino1,
Robert H.
Singer1,
Krishan L.
Taneja2,
Lawrence M.
Lifshitz3,
Ira M.
Herman4, and
Kenneth S.
Kosik5
1 Department of Anatomy and Structural Biology, Albert
Einstein College of Medicine, Bronx, New York 10461, 2 Department of Cell Biology and 3 Biomedical
Imaging Group, University of Massachusetts Medical Center, Worcester,
Massachusetts 10615, 4 Department of Physiology, Tufts
University School of Medicine, Boston, Massachusetts, and
5 Center for Neurological Disease, Brigham and Women's
Hospital, Harvard Medical School, Boston, Massachusetts
The transport of mRNAs into developing dendrites and axons may be a
basic mechanism to localize cytoskeletal proteins to growth cones and
influence microfilament organization. Using isoform-specific antibodies
and probes for in situ hybridization, we observed
distinct localization patterns for - and -actin within cultured
cerebrocortical neurons. -Actin protein was highly enriched within
growth cones and filopodia, in contrast to -actin protein, which was
distributed uniformly throughout the cell. -Actin protein also was
shown to be peripherally localized after transfection of -actin cDNA bearing an epitope tag. -Actin mRNAs were localized more frequently to neuronal processes and growth cones, unlike -actin mRNAs, which
were restricted to the cell body. The rapid localization of -actin
mRNA, but not -actin mRNA, into processes and growth cones could be
induced by dibutyryl cAMP treatment. Using high-resolution in
situ hybridization and image-processing methods, we showed that
the distribution of -actin mRNA within growth cones was statistically nonrandom and demonstrated an association with
microtubules. -Actin mRNAs were detected within minor neurites,
axonal processes, and growth cones in the form of spatially distinct
granules that colocalized with translational components.
Ultrastructural analysis revealed polyribosomes within growth cones
that colocalized with cytoskeletal filaments. The transport of
-actin mRNA into developing neurites may be a sequence-specific
mechanism to synthesize cytoskeletal proteins directly within processes
and growth cones and would provide an additional means to deliver
cytoskeletal proteins over long distances.
Key words:
mRNA localization; actin isoforms; cytoskeleton; growth
cones; axonal transport; in situ
hybridization
Copyright © 1998 Society for Neuroscience 0270-6474/98/181251-15$05.00/0
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R.-P. JANSEN
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I. Nabi
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E.-L. Punnonen, C. Fages, J. Wartiovaara, and H. Rauvala
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H. Eng, K. Lund, and R. B. Campenot
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L. Havin, A. Git, Z. Elisha, F. Oberman, K. Yaniv, S. P. Schwartz, N. Standart, and J. K. Yisraeli
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D. A. Potter, J. S. Tirnauer, R. Janssen, D. E. Croall, C. N. Hughes, K. A. Fiacco, J. W. Mier, M. Maki, and I. M. Herman
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P. M. C. Wong, Q. Yuan, H. Chen, B. M. Sultzer, and S.-W. Chung
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W. Gu, F. Pan, H. Zhang, G. J. Bassell, and R. H. Singer
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G. Liu, W. M. Grant, D. Persky, V. M. Latham Jr., R. H. Singer, and J. Condeelis
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