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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-04-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fnmol.2023.1125160/full |
| _version_ | 1797849273364119552 |
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| author | Harish R. Krishnan Gian Paolo Vallerini Hannah E. Gavin Marina Guizzetti Marina Guizzetti Hooriyah S. Rizavi Hooriyah S. Rizavi David P. Gavin David P. Gavin Rajiv P. Sharma Rajiv P. Sharma |
| author_facet | Harish R. Krishnan Gian Paolo Vallerini Hannah E. Gavin Marina Guizzetti Marina Guizzetti Hooriyah S. Rizavi Hooriyah S. Rizavi David P. Gavin David P. Gavin Rajiv P. Sharma Rajiv P. Sharma |
| author_sort | Harish R. Krishnan |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-09T18:42:21Z |
| format | Article |
| id | doaj.art-e8c583c98e94474d810827584b3aeb76 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-04-09T18:42:21Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-e8c583c98e94474d810827584b3aeb762023-04-11T04:45:49ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992023-04-011610.3389/fnmol.2023.11251601125160Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neuronsHarish R. Krishnan0Gian Paolo Vallerini1Hannah E. Gavin2Marina Guizzetti3Marina Guizzetti4Hooriyah S. Rizavi5Hooriyah S. Rizavi6David P. Gavin7David P. Gavin8Rajiv P. Sharma9Rajiv P. Sharma10Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Psychiatry, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Psychiatry, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Behavioral Neuroscience, Oregon Health & Science University, Portland, OR, United StatesVA Portland Health Care System, Portland, OR, United StatesDepartment of Psychiatry, University of Illinois at Chicago, Chicago, IL, United StatesJesse Brown Veterans Affairs Medical Center, Chicago, IL, United StatesDepartment of Psychiatry, University of Illinois at Chicago, Chicago, IL, United StatesJesse Brown Veterans Affairs Medical Center, Chicago, IL, United StatesDepartment of Psychiatry, University of Illinois at Chicago, Chicago, IL, United StatesJesse Brown Veterans Affairs Medical Center, Chicago, IL, United StatesWe 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.https://www.frontiersin.org/articles/10.3389/fnmol.2023.1125160/fullTRAP-seqribosomal engagementRNA sequencingalcohol use disorder (AUD)PARP (poly(ADP-ribose) polymeraseABT-888 |
| spellingShingle | Harish R. Krishnan Gian Paolo Vallerini Hannah E. Gavin Marina Guizzetti Marina Guizzetti Hooriyah S. Rizavi Hooriyah S. Rizavi David P. Gavin David P. Gavin Rajiv P. Sharma Rajiv P. Sharma Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons Frontiers in Molecular Neuroscience TRAP-seq ribosomal engagement RNA sequencing alcohol use disorder (AUD) PARP (poly(ADP-ribose) polymerase ABT-888 |
| 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 | TRAP-seq ribosomal engagement RNA sequencing alcohol use disorder (AUD) PARP (poly(ADP-ribose) polymerase ABT-888 |
| url | https://www.frontiersin.org/articles/10.3389/fnmol.2023.1125160/full |
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