The synaptoneurosome transcriptome: a model for profiling the emolecular effects of alcohol

Most, Dana; Ferguson, Laura B.; Blednov, Yuri A.; Mayfield, R. Dayne; Harris, R. Adron

doi:10.1038/tpj.2014.43

Cited by 31 publications

(54 citation statements)

References 73 publications

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“…Cell-type specific databases provide one tool to identify the cellular basis of the transcriptome changes by identifying the gene co-expression modules that are enriched in different cells. This strategy has also helped to identify discrete cells that are targeted depending on the ethanol drinking paradigms, time points and brain regions studied [111][112][113][114]. Because microglia make up only 5-15% of cells in the brain, it is likely that key microglial gene expression changes are missed when examining whole tissue [115], making it necessary to perform transcriptome studies in isolated microglia.…”

Section: Gene Expression In Isolated Microgliamentioning

confidence: 99%

The Neuroimmune Transcriptome and Alcohol Dependence: Potential for Targeted Therapies

et al. 2016

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Transcriptome profiling enables discovery of gene networks that are altered in alcoholic brains. This technique has revealed involvement of the brain's neuroimmune system in regulating alcohol abuse and dependence, and has provided potential therapeutic targets. In this review, we discuss Toll-like-receptor pathways, hypothesized to be key players in many stages of the alcohol addiction cycle. The growing appreciation of the neuroimmune system's involvement in alcoholism has also led to consideration of crucial roles for glial cells, including astrocytes and microglia, in the brain's response to alcohol abuse. We discuss current knowledge and hypotheses on the roles that specific neuroimmune cell types may play in addiction. Current strategies for repurposing US FDA-approved drugs for the treatment of alcohol use disorders are also discussed.

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Section: Gene Expression In Isolated Microgliamentioning

confidence: 99%

The Neuroimmune Transcriptome and Alcohol Dependence: Potential for Targeted Therapies

et al. 2016

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“…As described previously (Most et al, 2014), after 30 days of the two-bottle choice drinking, 8 alcohol-consuming and 12 control mice were killed by cervical dislocation and decapitated at the beginning of the light phase of the light/ dark cycle. Brains were removed and washed for 1 min with 1 ml of ice-cold homogenizing buffer (HB) containing 20 mM HEPES, 1 mM EDTA (pH 7.4), 40 U/ml RNAseOut (Invitrogen, CA), phosphatase inhibitor cocktail 3 (Sigma, MO) and protease inhibitors 'Complete' (Roche, IN).…”

Section: Rna Extractionmentioning

confidence: 99%

“…The supernatant was removed and the pellet was snap frozen and stored at − 80°C for SN RNA analysis. Microscopy was used to further characterize the SN preparation (Most et al, 2014).…”

Section: Rna Extractionmentioning

confidence: 99%

“…Studies investigating the effects of chronic alcohol consumption in standard tissue (total homogenate, TH) preparations found a persistent change in the expression of microRNAs and their target mRNAs in humans, mice, and rats (Lewohl et al, 2011;Gorini et al, 2013;Nunez et al, 2013;Tapocik et al, 2013). However, the standard TH preparation likely underestimates the number and magnitude of alcoholresponsive transcripts localized in the synapse (Lewohl et al, 2011;Most et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…We recently showed that alcohol-induced mRNA changes are greater in SN compared with TH (Most et al, 2014). Here, we profiled the microRNA transcriptomes from paired SN and TH samples of mouse amygdala after a voluntary alcohol consumption paradigm.…”

Section: Introductionmentioning

confidence: 99%

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Synaptic microRNAs Coordinately Regulate Synaptic mRNAs: Perturbation by Chronic Alcohol Consumption

Most

Leiter

Blednov

et al. 2015

Neuropsychopharmacol

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Local translation of mRNAs in the synapse has a major role in synaptic structure and function. Chronic alcohol use causes persistent changes in synaptic mRNA expression, possibly mediated by microRNAs localized in the synapse. We profiled the transcriptome of synaptoneurosomes (SN) obtained from the amygdala of mice that consumed 20% ethanol (alcohol) in a 30-day continuous two-bottle choice test to identify the microRNAs that target alcohol-induced mRNAs. SN are membrane vesicles containing pre-and post-synaptic compartments of neurons and astroglia and are a unique model for studying the synaptic transcriptome. We previously showed that chronic alcohol regulates mRNA expression in a coordinated manner. Here, we examine microRNAs and mRNAs from the same samples to define alcohol-responsive synaptic microRNAs and their predicted interactions with targeted mRNAs. The aim of the study was to identify the microRNA-mRNA synaptic interactions that are altered by alcohol. This was accomplished by comparing the effect of alcohol in SN and total homogenate preparations from the same samples. We used a combination of unbiased bioinformatic methods (differential expression, correlation, co-expression, microRNA-mRNA target prediction, co-targeting, and cell type-specific analyses) to identify key alcohol-sensitive microRNAs. Prediction analysis showed that a subset of alcohol-responsive microRNAs was predicted to target many alcohol-responsive mRNAs, providing a bidirectional analysis for identifying microRNA-mRNA interactions. We found microRNAs and mRNAs with overlapping patterns of expression that correlated with alcohol consumption. Cell type-specific analysis revealed that a significant number of alcohol-responsive mRNAs and microRNAs were unique to glutamate neurons and were predicted to target each other. Chronic alcohol consumption appears to perturb the coordinated microRNA regulation of mRNAs in SN, a mechanism that may explain the aberrations in synaptic plasticity affecting the alcoholic brain.

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Hippocampal transcriptome reveals novel targets of FASD pathogenesis

et al. 2019

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Introduction Prenatal alcohol exposure can contribute to fetal alcohol spectrum disorders (FASD), characterized by a myriad of developmental impairments affecting behavior and cognition. Studies show that many of these functional impairments are associated with the hippocampus, a structure exhibiting exquisite vulnerability to developmental alcohol exposure and critically implicated in learning and memory; however, mechanisms underlying alcohol‐induced hippocampal deficits remain poorly understood. By utilizing a high‐throughput RNA‐sequencing (RNA‐seq) approach to address the neurobiological and molecular basis of prenatal alcohol‐induced hippocampal functional deficits, we hypothesized that chronic binge prenatal alcohol exposure alters gene expression and global molecular pathways in the fetal hippocampus. Methods Timed‐pregnant Sprague–Dawley rats were randomly assigned to a pair‐fed control (PF) or binge alcohol (ALC) treatment group on gestational day (GD) 4. ALC dams acclimatized from GDs 5–10 with a daily treatment of 4.5 g/kg alcohol and subsequently received 6 g/kg on GDs 11–20. PF dams received a once daily maltose dextrin gavage on GDs 5–20, isocalorically matching ALC counterparts. On GD 21, bilateral hippocampi were dissected, flash frozen, and stored at −80°C. Total RNA was then isolated from homogenized tissues. Samples were normalized to ~4nM and pooled equally. Sequencing was performed by Illumina NextSeq 500 on a 75 cycle, single‐end sequencing run. Results RNA‐seq identified 13,388 genes, of these, 76 genes showed a significant difference (p < 0.05, log2 fold change ≥2) in expression between the PF and ALC groups. Forty‐nine genes showed sex‐dependent dysregulation; IPA analysis showed among female offspring, dysregulated pathways included proline and citrulline biosynthesis, whereas in males, xenobiotic metabolism signaling and alaninine biosynthesis etc. were altered. Conclusion We conclude that chronic binge alcohol exposure during pregnancy dysregulates fetal hippocampal gene expression in a sex‐specific manner. Identification of subtle, transcriptome‐level dysregulation in hippocampal molecular pathways offers potential mechanistic insights underlying FASD pathogenesis.

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The synaptoneurosome transcriptome: a model for profiling the emolecular effects of alcohol

Cited by 31 publications

References 73 publications

The Neuroimmune Transcriptome and Alcohol Dependence: Potential for Targeted Therapies

The Neuroimmune Transcriptome and Alcohol Dependence: Potential for Targeted Therapies

Synaptic microRNAs Coordinately Regulate Synaptic mRNAs: Perturbation by Chronic Alcohol Consumption

Hippocampal transcriptome reveals novel targets of FASD pathogenesis

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