Unknown,Transcriptomics,Genomics,Proteomics

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Alterations to the Synaptic Transcriptome in Response to Ethanol-Induced Behavioral Sensitization in DBA/2J Mice

ABSTRACT: Alcoholism is a complex neurological disorder characterized by loss of control in limiting intake, and progressive compulsion to seek and consume ethanol. Prior studies have suggested that the characteristic behaviors associated with escalation of drug use are caused, at least in part, by ethanol-evoked changes in gene expression affecting synaptic plasticity. Implicit in this hypothesis is a dependence on new protein synthesis and remodeling at the synapse. It is well established that mRNA can be transported to neuronal distal processes, where it can undergo localized translation that is regulated in a spatially restricted manner in response to stimulation. It is unknown whether such modulation of the synaptic transcriptome might contribute to ethanol-induced synaptic plasticity. Using ethanol-induced behavioral sensitization as a model of neuroplasticity, here we investigated whether repeated exposure to ethanol altered the synaptic transcriptome, contributing to synaptic plasticity underlying a subsequent increase in the ethanol-evoked locomotor response. A synaptoneurosome preparation was utilized to enrich for RNAs trafficked to the synapse versus the proximal cellular fraction. Two complementary methods of genomic profiling, RNA-Seq and microarrays, were used to survey the synaptic transcriptome of DBA/2J mice subjected to ethanol-induced behavioral sensitization. Strikingly, a large number of genes regulated by acute ethanol showed adaptation to repeated ethanol exposure with sensitization. Genomic profiling showed distinct functional classes in RNA found to be enriched in the synaptoneurosome faction, consistent with their role in synaptic function. A subset of ethanol-responsive genes significantly enriched at the synapse were related to biological functions such as protein folding and extra-cellular matrix components, suggesting a role for local regulation of synaptic protein function by ethanol. In contrast, RNA classes regulated by ethanol in the cell soma included an over-abundance of RNA-binding and trafficking proteins, perhaps reflecting increased demand for dendritic RNA localization. These experiments document that both acute and repeated ethanol produce specific alterations in the complement of RNA at the synapse and lay the foundation for further investigations into the role of the synaptic transcriptome in behavioral adaptations to ethanol. 24 samples were analyzed from 3 treatment groups and 2 cell fractions with 4 biological replicates for each treatment/cell fraction.Cell fractionation was carried out on tissue from a frontal pole dissection of DBA2/J mice, isolating a cytosolic (S2) and synaptoneurosomal (P2) fraction. Treatment groups describe chronic treatment (10 daily injections) and treatment used on test day (single injeciion), producing saline-saline (SS), saline-ethanol (SE) and ethanol-ethanol (EE) groups. Analysis of RNA-seq compared similar treatments across cell fractions or different treatments within a cell fraction.

ORGANISM(S): Mus musculus

SUBMITTER: Michael Miles

PROVIDER: E-GEOD-73018 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Alcoholism is a complex neurological disorder characterized by loss of control in limiting intake, and progressive compulsion to seek and consume ethanol. Prior studies have suggested that the characteristic behaviors associated with escalation of drug use are caused, at least in part, by ethanol-evoked changes in gene expression affecting synaptic plasticity. Implicit in this hypothesis is a dependence on new protein synthesis and remodeling at the synapse. It is well established that mRNA can be transported to neuronal distal processes, where it can undergo localized translation that is regulated in a spatially restricted manner in response to stimulation. It is unknown whether such modulation of the synaptic transcriptome might contribute to ethanol-induced synaptic plasticity. Using ethanol-induced behavioral sensitization as a model of neuroplasticity, here we investigated whether repeated exposure to ethanol altered the synaptic transcriptome, contributing to synaptic plasticity underlying a subsequent increase in the ethanol-evoked locomotor response. A synaptoneurosome preparation was utilized to enrich for RNAs trafficked to the synapse versus the proximal cellular fraction. Two complementary methods of genomic profiling, RNA-Seq and microarrays, were used to survey the synaptic transcriptome of DBA/2J mice subjected to ethanol-induced behavioral sensitization. Strikingly, a large number of genes regulated by acute ethanol showed adaptation to repeated ethanol exposure with sensitization. Genomic profiling showed distinct functional classes in RNA found to be enriched in the synaptoneurosome faction, consistent with their role in synaptic function. A subset of ethanol-responsive genes significantly enriched at the synapse were related to biological functions such as protein folding and extra-cellular matrix components, suggesting a role for local regulation of synaptic protein function by ethanol. In contrast, RNA classes regulated by ethanol in the cell soma included an over-abundance of RNA-binding and trafficking proteins, perhaps reflecting increased demand for dendritic RNA localization. These experiments document that both acute and repeated ethanol produce specific alterations in the complement of RNA at the synapse and lay the foundation for further investigations into the role of the synaptic transcriptome in behavioral adaptations to ethanol.

Project description:Histone acetylation and other modifications of the chromatin are important regulators of gene expression and, consequently, may contribute to drug-induced behaviors and neuroplasticity. Previous studies have shown that a reduction on histone deacetylase (HDAC) activity results on the enhancement of some psychostimulant-induced behaviors. In the present study, we extend those seminal findings by showing that the administration of the HDAC inhibitor sodium butyrate enhances morphine-induced locomotor sensitization and conditioned place preference. In contrast, this compound has no effects on the development of morphine tolerance and dependence. Similar effects were observed for cocaine and ethanol-induced behaviors. These behavioral changes were accompanied by a selective boosting of a component of the transcriptional program activated by chronic morphine administration that included circadian clock genes and other genes relevant in addictive behavior. Our results support an specific role for histone acetylation and the epigenetic modulation of transcription at a reduced number of biologically relevant loci on non-homeostatic, long lasting, drug-induced behavioral plasticity. To further investigate the molecular bases of sodium butyrate action on long-lasting behavioral responses to morphine, we screened for potential substrates of their interaction by performing a genome-wide comparison of the striatal transcriptome after chronic administration of morphine in the absence or presence of sodium butyrate. Striatal tissue was dissected from two months-old Swiss-Albino male mice subjected to behavioral sensitization. Behavioural sensitization: Locomotor sensitization induced by morphine was evaluated using a protocol divided into two phases: induction and challenge. The induction phase involved six trials on alternate days, one trial per day. On each one of these trials, mice received and injection of saline or sodium butyrate (300 mg/kg) immediatedly followed by a second injection of morphine (20 mg/kg). The challenge phases consisted of a single trial conducted 7 days after the last test of the induction phase. In this case, all animals received a single injection of morphine (20 mg/kg). Striatal RNA was extracted 1h after the morphine challenge in groups of four animals that received either saline (N=6) or morphine (N=3), or the sodium butyrate-morphine (N=3) co-treatment during the sensitization induction phase.

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Alterations to the Synaptic Transcriptome in Response to Ethanol-Induced Behavioral Sensitization in DBA/2J Mice

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