| Literature DB >> 34939924 |
Caryn R Hale1, Kirsty Sawicka1, Kevin Mora1, John J Fak1, Jin Joo Kang1, Paula Cutrim1, Katarzyna Cialowicz2, Thomas S Carroll3, Robert B Darnell1,4.
Abstract
Neurons rely on translation of synaptic mRNAs in order to generate activity-dependent changes in plasticity. Here, we develop a strategy combining compartment-specific crosslinking immunoprecipitation (CLIP) and translating ribosome affinity purification (TRAP) in conditionally tagged mice to precisely define the ribosome-bound dendritic transcriptome of CA1 pyramidal neurons. We identify CA1 dendritic transcripts with differentially localized mRNA isoforms generated by alternative polyadenylation and alternative splicing, including many that have altered protein-coding capacity. Among dendritic mRNAs, FMRP targets were found to be overrepresented. Cell-type-specific FMRP-CLIP and TRAP in microdissected CA1 neuropil revealed 383 dendritic FMRP targets and suggests that FMRP differentially regulates functionally distinct modules in CA1 dendrites and cell bodies. FMRP regulates ~15-20% of mRNAs encoding synaptic functions and 10% of chromatin modulators, in the dendrite and cell body, respectively. In the absence of FMRP, dendritic FMRP targets had increased ribosome association, consistent with a function for FMRP in synaptic translational repression. Conversely, downregulation of FMRP targets involved in chromatin regulation in cell bodies suggests a role for FMRP in stabilizing mRNAs containing stalled ribosomes in this compartment. Together, the data support a model in which FMRP regulates the translation and expression of synaptic and nuclear proteins within different compartments of a single neuronal cell type.Entities:
Keywords: FMRP; RNA-binding proteins; genetics; genomics; local translation; mRNA localization; mouse; neuroscience; synaptic plasticity
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Year: 2021 PMID: 34939924 PMCID: PMC8820740 DOI: 10.7554/eLife.71892
Source DB: PubMed Journal: Elife ISSN: 2050-084X Impact factor: 8.140