Effects of alcohol and PARP inhibition on RNA ribosomal engagement in cortical excitatory neurons
The objective of the study is to describe the effects of EtOH on excitatory pre-frontal cortical neuronal ribosomal-engagement (RE) transcripts from total-RNA and provides insights into Poly (ADP-ribose) polymerase (PARP)-mediated regulation of EtOH effects (using the PARP inhibitor ABT-888). Overall design: Mice whose ribosomes were tagged with EGFP expressed only in CaMKIIa expressing cells were used for the study. PFC samples were homogenized in the appropriate buffer and TRAP-RNA (collected by anti-EGFP antibody incubation) and total-RNA samples were extracted. Following quantification and quality control, the samples were subjected to rRNA depletion, library preparation and 50bp paired-end sequencing (NovaSeq 6000) using the Illumina platform. Data were analyzed using standardized bioinformatic packages and normalized based on treatment status, i.e., Control, EtOH or EtOH+ABT-888, as well as by sample type, i.e., Input or TRAP.