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Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons
We report on the effects of ethanol (EtOH) and Poly (ADP-ribose) polymerase (PARP) inhibition on RNA ribosomal engagement, as a proxy for protein translation, in prefrontal cortical (PFC) pyramidal neurons. We hypothesized that EtOH induces a shift in RNA ribosomal-engagement (RE) in PFC pyramidal n...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10126255/ https://www.ncbi.nlm.nih.gov/pubmed/37113267 http://dx.doi.org/10.3389/fnmol.2023.1125160 |
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author | Krishnan, Harish R. Vallerini, Gian Paolo Gavin, Hannah E. Guizzetti, Marina Rizavi, Hooriyah S. Gavin, David P. Sharma, Rajiv P. |
author_facet | Krishnan, Harish R. Vallerini, Gian Paolo Gavin, Hannah E. Guizzetti, Marina Rizavi, Hooriyah S. Gavin, David P. Sharma, Rajiv P. |
author_sort | Krishnan, Harish R. |
collection | PubMed |
description | We report on the effects of ethanol (EtOH) and Poly (ADP-ribose) polymerase (PARP) inhibition on RNA ribosomal engagement, as a proxy for protein translation, in prefrontal cortical (PFC) pyramidal neurons. We hypothesized that EtOH induces a shift in RNA ribosomal-engagement (RE) in PFC pyramidal neurons, and that many of these changes can be reversed using a PARP inhibitor. We utilized the translating ribosome affinity purification (TRAP) technique to isolate cell type-specific RNA. Transgenic mice with EGFP-tagged Rpl10a ribosomal protein expressed only in CaMKIIα-expressing pyramidal cells were administered EtOH or normal saline (CTL) i.p. twice a day, for four consecutive days. On the fourth day, a sub-group of mice that received EtOH in the previous three days received a combination of EtOH and the PARP inhibitor ABT-888 (EtOH + ABT-888). PFC tissue was processed to isolate both, CaMKIIα pyramidal cell-type specific ribosomal-engaged RNA (TRAP-RNA), as well as genomically expressed total-RNA from whole tissue, which were submitted for RNA-seq. We observed EtOH effects on RE transcripts in pyramidal cells and furthermore treatment with a PARP inhibitor “reversed” these effects. The PARP inhibitor ABT-888 reversed 82% of the EtOH-induced changes in RE (TRAP-RNA), and similarly 83% in the total-RNA transcripts. We identified Insulin Receptor Signaling as highly enriched in the ethanol-regulated and PARP-reverted RE pool and validated five participating genes from this pathway. To our knowledge, this is the first description of the effects of EtOH on excitatory neuron RE transcripts from total-RNA and provides insights into PARP-mediated regulation of EtOH effects. |
format | Online Article Text |
id | pubmed-10126255 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-101262552023-04-26 Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons Krishnan, Harish R. Vallerini, Gian Paolo Gavin, Hannah E. Guizzetti, Marina Rizavi, Hooriyah S. Gavin, David P. Sharma, Rajiv P. Front Mol Neurosci Neuroscience We report on the effects of ethanol (EtOH) and Poly (ADP-ribose) polymerase (PARP) inhibition on RNA ribosomal engagement, as a proxy for protein translation, in prefrontal cortical (PFC) pyramidal neurons. We hypothesized that EtOH induces a shift in RNA ribosomal-engagement (RE) in PFC pyramidal neurons, and that many of these changes can be reversed using a PARP inhibitor. We utilized the translating ribosome affinity purification (TRAP) technique to isolate cell type-specific RNA. Transgenic mice with EGFP-tagged Rpl10a ribosomal protein expressed only in CaMKIIα-expressing pyramidal cells were administered EtOH or normal saline (CTL) i.p. twice a day, for four consecutive days. On the fourth day, a sub-group of mice that received EtOH in the previous three days received a combination of EtOH and the PARP inhibitor ABT-888 (EtOH + ABT-888). PFC tissue was processed to isolate both, CaMKIIα pyramidal cell-type specific ribosomal-engaged RNA (TRAP-RNA), as well as genomically expressed total-RNA from whole tissue, which were submitted for RNA-seq. We observed EtOH effects on RE transcripts in pyramidal cells and furthermore treatment with a PARP inhibitor “reversed” these effects. The PARP inhibitor ABT-888 reversed 82% of the EtOH-induced changes in RE (TRAP-RNA), and similarly 83% in the total-RNA transcripts. We identified Insulin Receptor Signaling as highly enriched in the ethanol-regulated and PARP-reverted RE pool and validated five participating genes from this pathway. To our knowledge, this is the first description of the effects of EtOH on excitatory neuron RE transcripts from total-RNA and provides insights into PARP-mediated regulation of EtOH effects. Frontiers Media S.A. 2023-04-11 /pmc/articles/PMC10126255/ /pubmed/37113267 http://dx.doi.org/10.3389/fnmol.2023.1125160 Text en Copyright © 2023 Krishnan, Vallerini, Gavin, Guizzetti, Rizavi, Gavin and Sharma. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Neuroscience Krishnan, Harish R. Vallerini, Gian Paolo Gavin, Hannah E. Guizzetti, Marina Rizavi, Hooriyah S. Gavin, David P. Sharma, Rajiv P. Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons |
title | Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons |
title_full | Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons |
title_fullStr | Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons |
title_full_unstemmed | Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons |
title_short | Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons |
title_sort | effects of alcohol and parp inhibition on rna ribosomal engagement in cortical excitatory neurons |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10126255/ https://www.ncbi.nlm.nih.gov/pubmed/37113267 http://dx.doi.org/10.3389/fnmol.2023.1125160 |
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