Transcriptional analysis of the effects of early life exposure to fluoxetine on telencephalon and hypothalamus in SR4G transgenic zebrafish larvae and adults
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ABSTRACT:
PROVIDER: PRJNA592212 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR10550509.fastq.gz | Fastqsanger.gz | |||
| SRR10550510.fastq.gz | Fastqsanger.gz | |||
| SRR10550511.fastq.gz | Fastqsanger.gz | |||
| SRR10550512.fastq.gz | Fastqsanger.gz | |||
| SRR10550515.fastq.gz | Fastqsanger.gz |
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Project description:Our previous study suggested persistent transgenerational effects of fluoxetine (FLX) exposure, in both environmental and therapeutic concentrations, on stress response and behaviour up to 3 generation withdrawn from exposure in larvae and adult zebrafish. To lay the foundations of the mechanistic of such transgenerational effects, we studied the effect of FLX on stress response in brain tissues (telencephalon and hypothalamus) of larvae and adult zebrafish using differentially expressed genes at the transcriptome level. To monitor the stress response, we used a transgenic zebrafish line (SR4G) which are expressing green fluorescence protein (GFP) in response to endogenous and exogenous glucocorticoids. The transcriptome analysis of GFP RNA as an internal control, provided us with an invaluable tool to clearly visualize and differentiate between stressed and unstressed groups as per treatments. To stimulate stress in both larvae and adult zebrafish we used a modified net stressing technique consisting of consecutive sequence of air exposure and incubation intervals. A pool of 22 larvae heads and a pool of 5 telencephalon and 5 hypothalamus dissected tissues were used as the target tissues for RNA extraction. All experiments were replicated in n=6 per 4 different groups including Control-Unstressed (Cnt-U), Control-Stressed (Cnt-S), Fluoxetine-Unstressed (FLX-U), and Fluoxetine-Stressed (FLX-S) using pools of larvae heads, telencephalon, and hypothalamus. After total RNA extraction, using Illumina technology all samples were sequence and the list of differentially expressed genes (DEGs) were produced using DSEQ pipeline in R.
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Project description:Sexually mature (mid-May) adult female goldfish received a single injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 50 ug/g body weight at 5 uL/g body weight) or 0.6% saline on day 0 and a single injection of alpha-methyl-p-tyrosine (aMPT; 240 ug/g body weight at 5 uL/g body weight) or 0.6% saline on Day 5 with the goal of severely deplete catecholamine (dopamine and norepinephrine) levels. Hypothalamus tissue was extracted on Day 6. Telencephalon tissue was extracted on Day 6. The objective of this study was to examine gene expression changes in the neuroendocrine brain in response to catecholamine depletion. A common reference design was used for both the hypothalamus and telencephalon groups: Hypothalamus: 3 independent groups of hypothalami from sexually mature female goldfish treated with 50 ug/g MPTP + 240 ug/g aMPT hybridized against a common vehicle control. An additional group was used for a dye-swap. Slides 4, 17, 21, 52. Telencephalon: 3 independent groups of telencephali from sexually mature female goldfish treated with 50 ug/g MPTP + 240 ug/g aMPT hybridized against a common vehicle control. An additional group was used for a dye-swap. Slides 28, 34, 43, 48.
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Project description:Sexually mature (mid-May) adult female goldfish received a single injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 50 ug/g body weight at 5 uL/g body weight) or 0.6% saline on day 0 and a single injection of alpha-methyl-p-tyrosine (aMPT; 240 ug/g body weight at 5 uL/g body weight) or 0.6% saline on Day 5 with the goal of severely deplete catecholamine (dopamine and norepinephrine) levels. Hypothalamus tissue was extracted on Day 6. Telencephalon tissue was extracted on Day 6. The objective of this study was to examine gene expression changes in the neuroendocrine brain in response to catecholamine depletion.
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2013-01-30 | E-GEOD-41076 | biostudies-arrayexpress
Project description:A goldfish brain enriched cDNA combined with a carp microarray (Gracey et al., 2004). Female adult goldish were treated twice a week for 17 days, for a total of five injections. Each of the injections consisted of 5 μg fluoxetine/g of body weight with an injection volume of 1μl/g. The telencephalon was tissue used. Keywords: i.p.injection of muscimol, 6 hours, pharmacological 4 slides (n=3 treatment samples and common control pool) slide 19 = treatment sample 1 slide 30 = treatment sample 2 slide 36 = treatment sample 3 (dye swap) slide 39 = treatment sample 3 (dye swap)
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Project description:Pharmaceutical chemicals used in human medicine are released into surface waters via municipal effluents and pose a risk for aquatic organisms. Among these substances are selective serotonin reuptake inhibitors (SSRIs) which can affect aquatic organisms at sub ppb concentrations. To better understand biochemical pathways influenced by SSRIs, evaluate changes in the transcriptome, and identify gene transcripts with potential for biomarkers of exposure to SSRIs; larval zebrafish Danio rerio were exposed (96 h) to two concentrations (25 and 250 µg/L) of the SSRIs, fluoxetine and sertraline, and changes in global gene expression were evaluated (Affymetrix GeneChip® Zebrafish Array). Significant changes in gene expression (>=1.7 fold change, p<0.05) were determined with Partek® Genomics Suite Gene Expression Data Analysis System and ontology analysis was conducted using Molecular Annotation System 3. The number of genes differentially expressed after fluoxetine exposure was 288 at 25 µg/L and 131 at 250 µg/L; and after sertraline exposure was 33 at 25 µg/L and 52 at 250 µg/L. Five genes were differentially regulated in all treatments relative to control, suggesting that both SSRIs share some similar molecular pathways. Among them, expression of the gene coding for FK506 binding protein 5 (FKBP5), which is annotated to stress response regulation, was highly down-regulated in all treatments (results confirmed by qRT-PCR). Gene ontology analysis indicated that regulation of stress response and cholinesterase activity were critical functions influenced by these SSRIs, and suggested that changes in the transcription of FKBP5 or acetylcholinesterase could be useful biomarkers of SSRIs exposure in wild fish. Zebrafish (Danio rerio) were obtained from the Zebrafish Research Facility maintained at the Center for Environmental Biotechnology at the University of Tennessee. Fish husbandry, spawning, and experimental procedures were conducted with approval from the UT Insititutional Animal Care and Use Committee (Protocol #1690-1007). Water for holding fish and conducting experiments (hereafter referred to as fish water) consisted of MilliQ water (Millipore, Bedford, MA) with ions added: 19 mg/L NaHCO3, 1 mg/L sea salt (Instant Ocean Synthetic Sea Salt, Mentor, OH), 10 mg/L CaSO4, 10 mg/L MgSO4, 2 mg/L KCl. Embroyos were obtained by spawning adult fish with no history of contaminant exposure. Fertilization of embryos took place at the same time (<15 minutes), such that larvae used in experiments were of similar age at the time of exposure. All activities (maintenance of adult fish, spawning, and experiments) were conducted in an environmental chamber with a temperature of 27 +/- 1 C and 14:10h light:dark photoperiod. Larval zebrafish (72 hpf) were exposed for 96 h in 200ml fish water containing appropirate amount of SSRI stock (i.e. fluoxetine or sertraline). There were four SSRIs treatments (25 and 250 ug/L fluoxetine and 25 and 250 ug/L sertraline) and one control (no SSRIs) with triplicate beakers and each beaker contained about 100 larval fish. During exposure for 96 hours, beakers were kept covered to prevent water evaporation and fish were not fed (i.e., fish consumed their yolk sac).
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Project description:We constructed a comprehensive multi-omics map of the molecular effects of fluoxetine (an SSRI antidepressant), in 27 rat brain regions. We profiled gene expression (bulk RNA-seq, 210 datasets) and chromatin state (bulk chromatin immunoprecipitation sequencing (ChIP-seq) for the histone marker H3K27ac, 100 datasets) in a broad, unbiased panel of 27 brain regions across the entire rodent brain, in naive and fluoxetine-treated animals. We complemented this approach with single-cell RNA-seq (scRNA-seq) analysis of two brain regions. Using diverse integrative data analysis techniques we characterized the complex and multifaceted effects of fluoxetine on region-specific and cell-type-specific gene regulatory networks and pathways. Remarkably, we observed profound molecular changes across the brain (>4,000 differentially expressed genes and differentially acetylated ChIP-seq peaks each) that were highly region-dependent. We leveraged this atlas to identify fluoxetine-moduated genes and gene-regulatory loci, predict enriched motifs that suggest potential upstream regulators, and validate global mechanisms of fluoxetine action.
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