Regulation of dopamine-dependent transcription and cocaine action by<i>Gadd45b</i>

Zipperly, Morgan E.; Sultan, Faraz; Graham, Guan-En; Brane, Andrew C.; Simpkins, Natalie A.; Ianov, Lara; Day, Jeremy J.

doi:10.1101/2020.05.01.072926

Cited by 4 publications

(3 citation statements)

References 68 publications

(120 reference statements)

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“…Two important caveats of the study are that we only examined males and at only one time point. Future studies should examine whether sex-dependent and/or estrus cycle-dependent effects on transcription occur within these paradigms [ 52 , 53 ], and different time points, as cocaine is known to differentially affect gene expression in a time-dependent manner [ 3 , 22 , 54 ]. It is also important to consider the variability in cocaine dosing within IVSA mice, as mice did not consistently self-administer 20 mg/kg dose in comparison to the other paradigms.…”

Section: Discussionmentioning

confidence: 99%

Cocaine induces paradigm-specific changes to the transcriptome within the ventral tegmental area

Campbell

Chen

Beardwood

et al. 2021

Neuropsychopharmacol.

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During the initial stages of drug use, cocaine-induced neuroadaptations within the ventral tegmental area (VTA) are critical for drug-associated cue learning and drug reinforcement processes. These neuroadaptations occur, in part, from alterations to the transcriptome. Although cocaine-induced transcriptional mechanisms within the VTA have been examined, various regimens and paradigms have been employed to examine candidate target genes. In order to identify key genes and biological processes regulating cocaine-induced processes, we employed genome-wide RNA-sequencing to analyze transcriptional profiles within the VTA from male mice that underwent one of four commonly used paradigms: acute home cage injections of cocaine, chronic home cage injections of cocaine, cocaine-conditioning, or intravenous-self administration of cocaine. We found that cocaine alters distinct sets of VTA genes within each exposure paradigm. Using behavioral measures from cocaine self-administering mice, we also found several genes whose expression patterns corelate with cocaine intake. In addition to overall gene expression levels, we identified several predicted upstream regulators of cocaine-induced transcription shared across all paradigms. Although distinct gene sets were altered across cocaine exposure paradigms, we found, from Gene Ontology (GO) term analysis, that biological processes important for energy regulation and synaptic plasticity were affected across all cocaine paradigms. Coexpression analysis also identified gene networks that are altered by cocaine. These data indicate that cocaine alters networks enriched with glial cell markers of the VTA that are involved in gene regulation and synaptic processes. Our analyses demonstrate that transcriptional changes within the VTA depend on the route, dose and context of cocaine exposure, and highlight several biological processes affected by cocaine. Overall, these findings provide a unique resource of gene expression data for future studies examining novel cocaine gene targets that regulate drug-associated behaviors.

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Section: Discussionmentioning

confidence: 99%

Cocaine induces paradigm-specific changes to the transcriptome within the ventral tegmental area

Campbell

Chen

Beardwood

et al. 2021

Neuropsychopharmacol.

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“…Information related to CRISPR sgRNA sequences, genomic targets, targeting strand, distance from the gene TSS, original design strategy (CRISPRa or CRISPRi), and potential off-target sites identified from Cas-OFFinder (Bae et al, 2014 ) is presented in Supplementary Table 1 . shRNA construction followed a similar approach, utilizing the Broad TRC shRNA design tool ( http://portals.broadinstitute.org/gpp/public/ ) and inserted into a lenti U6-shRNA/EF1α-mCherry vector (Zipperly et al, 2020 ). shRNAs were assessed for specificity using BLAST.…”

Section: Methodsmentioning

confidence: 99%

An Improved CRISPR/dCas9 Interference Tool for Neuronal Gene Suppression

et al. 2020

Self Cite

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The expression of genetic material governs brain development, differentiation, and function, and targeted manipulation of gene expression is required to understand contributions of gene function to health and disease states. Although recent improvements in CRISPR/dCas9 interference (CRISPRi) technology have enabled targeted transcriptional repression at selected genomic sites, integrating these techniques for use in non-dividing neuronal systems remains challenging. Previously, we optimized a dual lentivirus expression system to express CRISPR-based activation machinery in post-mitotic neurons. Here we used a similar strategy to adapt an improved dCas9-KRAB-MeCP2 repression system for robust transcriptional inhibition in neurons. We find that lentiviral delivery of a dCas9-KRAB-MeCP2 construct driven by the neuron-selective human synapsin promoter enabled transgene expression in primary rat neurons. Next, we demonstrate transcriptional repression using CRISPR sgRNAs targeting diverse gene promoters, and show superiority of this system in neurons compared to existing RNA interference methods for robust transcript specific manipulation at the complex Brain-derived neurotrophic factor (Bdnf) gene. Our findings advance this improved CRISPRi technology for use in neuronal systems for the first time, potentially enabling improved ability to manipulate gene expression states in the nervous system.

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“…One mechanism by which D1R-dependent signalling can stimulate IEG transcription is through CREB (cAMP response element binding protein) (17,18), which is activated by PKA and other downstream protein kinases (19)(20)(21). Here, however, CREB is only one player among many; D1R signalling converges on numerous transcription factors (17,(22)(23)(24) and chromatin regulators (25)(26)(27)(28)(29)(30) to control gene expression, and the specific mechanisms by which D1R signalling interacts with transcriptional regulators are only partially known. In the present study, we describe a role for bromodomain-containing protein 4 (Brd4) as a mediator of D1R-and cAMP/PKA-dependent transcription in striatal neurons.…”

Section: Introductionmentioning

confidence: 99%

Dopamine D1 receptor activation and cAMP/PKA signalling mediate Brd4 recruitment to chromatin to regulate gene expression in rat striatal neurons

Jones-Tabah

Chen

et al. 2021

Preprint

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The activity of striatal medium-spiny projection neurons is regulated by dopamine acting principally at D1 and D2 dopamine receptors. The dopamine D1 receptor (D1R) is a Gαs/olf-coupled GPCR which activates a cAMP/PKA/DARPP-32 signalling cascade that increases excitability and facilitates plasticity, partly through the regulation of transcription. Transcriptional regulation downstream of the D1R involves the activation of PKA, which can translocate to the nucleus to phosphorylate various targets. The chromatin reader Brd4 regulates transcription induced by neurotrophic factors in cortical neurons and has also been implicated in dopamine-dependent striatal functions. Brd4 is activated by phosphorylation; this facilitates its binding to acetylated histones at promoters and enhancers. In non-neuronal cells, Brd4 is recruited to chromatin in response to PKA signalling. However, it is unknown whether Brd4 is involved in transcriptional activation by the D1R in striatal neurons. Here, we demonstrate that cAMP/PKA signalling increases Brd4 recruitment to dopamine-induced genes in striatal neurons, and that knockdown or inhibition of Brd4 modulated D1R-induced gene expression. Specifically, inhibition of Brd4 with the bromodomain inhibitor JQ1 suppressed the expression of ~25% of D1R-upregulated genes, while increasing the expression of a subset of immediate-early genes, including Fos and Jun. This pro-transcriptional effect of JQ1 was P-TEFb-dependent, and mediated through inhibition of the BD1 bromodomain of Brd4. Finally, we report that JQ1 treatment downregulated expression of many GPCRs and also impaired ERK1/2 signalling in striatal neurons. Our findings identify Brd4 as a novel regulator of D1R-dependent transcription and delineate complex bi-directional effects of bromodomain inhibitors on neuronal transcription.

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Regulation of dopamine-dependent transcription and cocaine action byGadd45b

Cited by 4 publications

References 68 publications

Cocaine induces paradigm-specific changes to the transcriptome within the ventral tegmental area

Cocaine induces paradigm-specific changes to the transcriptome within the ventral tegmental area

An Improved CRISPR/dCas9 Interference Tool for Neuronal Gene Suppression

Dopamine D1 receptor activation and cAMP/PKA signalling mediate Brd4 recruitment to chromatin to regulate gene expression in rat striatal neurons

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