Journal
NATURE NEUROSCIENCE
Volume 15, Issue 12, Pages 1742-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nn.3246
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Funding
- US National Institutes of Health (NIH)
- US National Science Foundation
- Lucille Packard Children's Hospital Pediatric Research Fund
- Korea Research Foundation of the South Korea Ministry of Education, Science and Technology [R31-2008-000-10083-0]
- Howard Hughes Medical Institute
- NIH [4R37NS027177-23, P41GM103412-24, R01NS076860]
- Burroughs Wellcome Fund
- Rita Allen Foundation
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Protein synthesis is highly regulated throughout nervous system development, plasticity and regeneration. However, tracking the distributions of specific new protein species has not been possible in living neurons or at the ultrastructural level. Previously we created TimeSTAMP epitope tags, drug-controlled tags for immunohistochemical detection of specific new proteins synthesized at defined times. Here we extend TimeSTAMP to label new protein copies by fluorescence or photo-oxidation. Live microscopy of a fluorescent TimeSTAMP tag reveals that copies of the synaptic protein PSD95 are synthesized in response to local activation of growth factor and neurotransmitter receptors, and preferentially localize to stimulated synapses in rat neurons. Electron microscopy of a photo-oxidizing TimeSTAMP tag reveals new PSD95 at developing dendritic structures of immature neurons and at synapses in differentiated neurons. These results demonstrate the versatility of the TimeSTAMP approach for visualizing newly synthesized proteins in neurons.
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