FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons
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 precis...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-12-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/71892 |
| _version_ | 1811253444359487488 |
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| author | Caryn R Hale Kirsty Sawicka Kevin Mora John J Fak Jin Joo Kang Paula Cutrim Katarzyna Cialowicz Thomas S Carroll Robert B Darnell |
| author_facet | Caryn R Hale Kirsty Sawicka Kevin Mora John J Fak Jin Joo Kang Paula Cutrim Katarzyna Cialowicz Thomas S Carroll Robert B Darnell |
| author_sort | Caryn R Hale |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T16:51:00Z |
| format | Article |
| id | doaj.art-a7355e8c3b06440880d93b72cabe9183 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:51:00Z |
| publishDate | 2021-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-a7355e8c3b06440880d93b72cabe91832022-12-22T03:24:24ZengeLife Sciences Publications LtdeLife2050-084X2021-12-011010.7554/eLife.71892FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neuronsCaryn R Hale0Kirsty Sawicka1https://orcid.org/0000-0003-4195-6327Kevin Mora2John J Fak3Jin Joo Kang4Paula Cutrim5Katarzyna Cialowicz6Thomas S Carroll7Robert B Darnell8https://orcid.org/0000-0002-5134-8088Laboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United StatesLaboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United StatesLaboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United StatesLaboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United StatesLaboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United StatesLaboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United StatesBio-Imaging Resource Center, The Rockefeller University, New York, United StatesBioinformatics Resource Center, The Rockefeller University, New York, United StatesLaboratory of Molecular Neuro-Oncology, Rockefeller University, New York, United States; Howard Hughes Medical Institute, Rockefeller University, New York, United StatesNeurons 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.https://elifesciences.org/articles/71892local translationFMRPRNA-binding proteinssynaptic plasticitymRNA localization |
| spellingShingle | Caryn R Hale Kirsty Sawicka Kevin Mora John J Fak Jin Joo Kang Paula Cutrim Katarzyna Cialowicz Thomas S Carroll Robert B Darnell FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons eLife local translation FMRP RNA-binding proteins synaptic plasticity mRNA localization |
| title | FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons |
| title_full | FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons |
| title_fullStr | FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons |
| title_full_unstemmed | FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons |
| title_short | FMRP regulates mRNAs encoding distinct functions in the cell body and dendrites of CA1 pyramidal neurons |
| title_sort | fmrp regulates mrnas encoding distinct functions in the cell body and dendrites of ca1 pyramidal neurons |
| topic | local translation FMRP RNA-binding proteins synaptic plasticity mRNA localization |
| url | https://elifesciences.org/articles/71892 |
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