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Volume 16, Number 24, Issue of December 15, 1996 pp. 7812-7820
Copyright ©1996 Society for Neuroscience

Translocation of RNA Granules in Living Neurons

Received July 10, 1996; revised Sept. 13, 1996; accepted Sept. 24, 1996.

Roger B. Knowles1, James H. Sabry2, Maryann E. Martone3, Thomas J. Deerinck3, Mark H. Ellisman3, Gary J. Bassell1, and Kenneth S. Kosik1

1 Center for Neurological Diseases, Brigham and Women's Hospital, Boston, Massachusetts 02115, 2 Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, and 3 Microscopy and Imaging Resource, University of California, San Diego, La Jolla, California 92093-0608

Sorting of RNAs to specific subcellular loci occurs in diverse settings from fly oocytes to mammalian neurons. Using the membrane-permeable nucleic acid stain SYTO 14, we directly visualized the translocation of endogenous RNA in living cells. Labeled RNA was distributed nonrandomly as discrete granules in neuronal processes. The labeled granules colocalized with poly(A+) mRNA, with the 60S ribosomal subunit, and with elongation factor 1alpha , suggesting that granules represent a translational unit. A subset of labeled granules colocalized with beta -actin mRNA. Correlative light and electron microscopy indicated that the fluorescent granules corresponded to clusters of ribosomes at the ultrastructural level. Poststaining of sections with heavy metals confirmed the presence of ribosomes within these granules. In living neurons, a subpopulation of RNA granules was motile during the observation period. They moved at an average rate of 0.1 µm/sec. In young cultures their movements were exclusively anterograde, but after 7 d in culture, one-half of the motile granules moved in the retrograde direction. Granules in neurites were delocalized after treatment with microtubule-disrupting drugs. These results raise the possibility of a cellular trafficking system for the targeting of RNA in neurons.

Key words: RNA granules; cytoskeleton; ribosomes; beta -actin mRNA; elongation factor 1alpha ; photo-oxidation; RNA translocation




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