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Impact of the dopamine transporter knockdown in human colorectal cancer cells

ABSTRACT: Here, we performed transcriptome-profiling experiments on control scramble shRNA and shSLC6A3 HCT116 cells to study the impact DAT silencing on gene networks associated with neoplastic stemness. We also treated control HCT116 cells with the DAT agonist vanoxerine (VXN) to identify commonly regulated networks between DAT knockdown and pharmacological antagonism.

ORGANISM(S): Homo sapiens

PROVIDER: GSE226324 | GEO | 2023/11/17

REPOSITORIES: GEO

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Transcriptional response to amphetamine in Drosophila dopamine transporter (DAT) loss-of-function mutant

Project description:We used Drosophila genetic and behavioral models to examine AMPH-induced transcriptional changes in DAT-dependent manner, as those would be the most relevant to the stimulatory effects of the drug in the brain. We previously showed that flies respond to AMPH by increasing their locomotor activity and decreasing their sleep in a dopamine-dependent manner. Flies that carry a loss-of-function mutation in the gene encoding the Drosophila DAT homolog (dDATfmn, henceforth referred to as DAT mutants) exhibit heightened activity levels at baseline, consistent with increased levels of extracellular dopamine caused by the impairment of reuptake. In this study we compared gene expression changes in response to AMPH in brains of isogenic w1118 strain (WT) and DAT mutants. We found genes involved in the control of mRNA translation to be significantly upregulated in response to AMPH in a DAT-dependent manner.

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Characterizing the diversity of enteric neurons using Dopamine Transporter (DAT)-Cre reporter mice

Project description:Dopaminergic (DA) neurons are the predominant cell type in the midbrain that synthesize dopamine, a neurotransmitter implicated in various behavioural processes, including motor function, the reward pathway, and satiety. In diseases affecting these neurons, such as in Parkinson’s disease (PD), there is growing evidence that the gut-brain axis and selective vulnerability of DA neurons plays a crucial role in disease. Most investigations relating to DA neurons in the gut rely on immunoreactivity to tyrosine hydroxylase (TH) - a rate-limiting enzyme in the production of dopamine. However, the reliability of TH staining as a marker of DA neurons has been questioned in recent years. Our aim is to perform a comprehensive characterization of DA neurons in the gut using a well-accepted reporter mouse line, expressing a fluorescent protein under the dopamine transporter promoter (DAT). Our findings confirm a unique localization of DA neurons in the gut, and unveil that there are discrete subtypes of DA neurons in the gut, which we characterized using both immunofluorescence and single-cell transcriptomics. We observed distinct subtypes of DAT neurons expressing co-transmitters and modulators across both plexuses; some of them likely co-releasing acetylcholine, and a smaller population likely releasing nitric oxide; while others were positive for a slew of canonical DA markers (Vmat2, Girk2, Foxa2). Given the clear heterogeneity of DA gut neurons, further investigation is warranted to define their functional signatures and discover their inherent biological differences that predispose these cells to neurodegeneration.

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Transcriptional response to amphetamine in Drosophila dopamine transporter (DAT) loss-of-function mutant

Project description:Transcriptional response to amphetamine in Drosophila dopamine transporter (DAT) loss-of-function mutant

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Project description:We sought to find out the molecular mechanism of CDK5 displayed in colorectal cancer. Microarray experiments were carried out to identify different gene expression between CDK5-knockdown HCT116 cell lines and Scramble HCT116 cell lines.

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Project description:Midbrain dopamine (mDA) neurons play a central role in reward signaling and are widely implicated in psychiatric and neurodegenerative disorders. To understand how mDA neurons perform these functions, it is important to understand how mDA-specific genes are regulated. However, cellular heterogeneity in the mammalian brain presents a major challenge to obtaining this understanding. To this end, we developed a virus-based approach to label and capture mDA nuclei for transcriptome (RNA-Seq), and low-input chromatin accessibility (liDNase-Seq) profiling, followed by predictive modeling to identify putative transcriptional regulators of mDA neurons. Using this method, we identified Gmeb1, a transcription factor predicted to regulate expression of Th and Dat, genes critical for dopamine synthesis and reuptake, respectively. Gmeb1 knockdown in mDA neurons resulted in downregulation of Th and Dat, as well as in severe motor deficits. This study thus identifies Gmeb1 as a master regulator of mDA gene expression and function, and provides a general method for identifying cell type-specific transcriptional regulators..

2019-05-14 | GSE106956 | GEO

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