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The Journal of Neuroscience, September 1, 2000, 20(17):6385-6393
CaMKII 3' Untranslated Region-Directed mRNA Translocation in
Living Neurons: Visualization by GFP Linkage
Martha S.
Rook,
Mei
Lu, and
Kenneth S.
Kosik
Center for Neurological Diseases, Brigham and Women's Hospital,
Boston, Massachusetts 02115, and Department of Neurology, Harvard
Medical School, Boston, Massachusetts 02115
The CaMKII mRNA extends into distal hippocampal dendrites, and
the 3' untranslated region (3'UTR) is sufficient to mediate this
localization. We labeled the 3'UTR of the CaMKII mRNA in hippocampal
cultures by using a green fluorescent protein (GFP)/MS2 bacteriophage
tagging system. The CaMKII 3'UTR formed discrete granules throughout
the dendrites of transfected cells. The identity of the fluorescent
granules was verified by in situ hybridization. Over 30 min time periods these granules redistributed without a net increase in
granule number; with depolarization there is a tendency toward
increased numbers of granules in the dendrites. These observations
suggest that finer time resolution of granule motility might reveal
changes in the motility characteristics of granules after
depolarization. So that motile granules could be tracked, shorter
periods of observation were required. The movements of motile granules
can be categorized as oscillatory, unidirectional anterograde, or
unidirectional retrograde. Colocalization of CaMKII 3'UTR granules
and synapses suggested that oscillatory movements allowed the granules
to sample several local synapses. Neuronal depolarization increased the
number of granules in the anterograde motile pool. Based on the time
frame over which the granule number increased, the translocation of
granules may serve to prepare the dendrite for mounting an adequate
local translation response to future stimuli. Although the resident
pool of granules can respond to signals that induce local translation,
the number of granules in a dendrite might reflect its activation history.
Key words:
RNA localization; RNA translocation; CaMKII; 3'UTR; RNA granules; synaptic plasticity
Copyright © 2000 Society for Neuroscience 0270-6474/00/20176385-09$05.00/0
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