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The Journal of Neuroscience, October 15, 2003, 23(28):9409-9417
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Cellular/Molecular
An Unbiased cDNA Library Prepared from Isolated Aplysia Sensory Neuron Processes Is Enriched for Cytoskeletal and Translational mRNAs
Robert Moccia,1 * Dillon Chen,2 * Vlasta Lyles,1 Estreya Kapuya,2 Yaping E,4 Sergey Kalachikov,5 Christian M. T. Spahn,6 Joachim Frank,6 Eric R. Kandel,4 Mark Barad,1,2 andKelsey C. Martin1,2,3 1Brain Research Institute/Interdepartmental Program in Neuroscience, 2Department of Psychiatry and Biobehavioral Sciences, Neuropsychiatric Institute, 3Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, California 90095, 4Howard Hughes Medical Institute, Columbia University, Center for Neurobiology and Behavior, 5Columbia Genome Center, Columbia University, New York, New York 10032, and 6Howard Hughes Medical Institute, Health Center Inc., Albany, New York 12201
Local protein synthesis is required for long-lasting synapse-specific plasticity in cultured Aplysia sensorimotor synapses. To identify synaptically localized mRNAs, we prepared a cDNA library from isolated sensory neurites. By sequence analysis, we estimate that the library contains 263 distinct mRNAs, with 98 of these mRNAs constituting 70% of all clones. The localized transcripts are enriched for mRNAs encoding cytoskeletal elements and components of the translational machinery. In situ hybridization confirms that the mRNAs for at least eight of these transcripts are present in distal neurites. Immunocytochemistry reveals that serotonin regulates the translation of one of the localized mRNAs, that encoding
1-tubulin. Our identification of mRNAs encoding cytoskeletal elements suggests that local protein synthesis is required for the growth of new synaptic connections associated with persistent synaptic strengthening. Our finding of mRNAs encoding components of the translational machinery suggests that local protein synthesis serves to increase the translational capacity of synapses.
Key words: synaptic plasticity; mRNA localization; translation; synaptic tagging; Aplysia; cytoskeleton
Received Feb 26, 2003; revised July 18, 2003; accepted August 18, 2003.
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