MRNA-Seq of Danio rerio exposed to different concentrations of Fulvestrant against untreated control groups
Ontology highlight
ABSTRACT: In the present study transcriptome analysis was employed to investigate the early molecular responses to exposure to Fulvestrant (CAS 129453-61-8). Zebrafish embryos were exposed to Fulvestrant according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), simultaneous RNA and protein extraction from 10 embryos was performed using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system System and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential substance specific biomarker candidates were refined and selected based on the differential expression patterns and the biological functions investigation of the detected DEGs.
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to exposure to Tebuconazole (CAS 107534-96-3). Zebrafish embryos were exposed to Tebuconazole according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), simultaneous RNA and protein extraction from 10 embryos was performed using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system System and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential substance specific biomarker candidates were selected based on the differential expression patterns and the biological functions investigation of the detected DEGs.
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to exposure to 17β-estradiol (CAS 50-28-2). Zebrafish embryos were exposed to 17β-estradiol according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), simultaneous RNA and protein extraction from 10 embryos was performed using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system System and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential substance specific biomarker candidates were refined and selected based on the differential expression patterns and the biological functions investigation of the detected DEGs.
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to exposure to bisphenol A (CAS 80-05-7). Zebrafish embryos were exposed to to bisphenol A according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), simultaneous RNA and protein extraction from 10 embryos was performed using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system System and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential substance specific biomarker candidates were refined and selected based on the differential expression patterns and the biological functions investigation of the detected DEGs.
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to Iopanoic acid (CAS 96-83-3), a deiodinase inhibitor. Zebrafish embryos were exposed to Iopanoic acid according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), RNA was extracted from 10 embryos using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system (Illumina Inc., San Diego, USA) and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential thyroid disruption specific biomarker candidates were selected based on the differential expression patterns and the biological functions investigation of the detected differentially expressed genes (DEGs).
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to Methimazole (CAS 60-56-0), a thyroid peroxidase inhibitor. Zebrafish embryos were exposed to methimazole according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), RNA was extracted from 10 embryos using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system (Illumina Inc., San Diego, USA) and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential thyroid disruption specific biomarker candidates were selected based on the differential expression patterns and the biological functions investigation of the detected differentially expressed genes (DEGs).
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to exposure to epoxiconazole (CAS 133855-98-8). Zebrafish embryos were exposed to epoxiconazole according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), simultaneous RNA and protein extraction from 10 embryos was performed using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina NovaSeq 6000 system System and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential substance specific biomarker candidates were selected based on the differential expression patterns and the biological functions investigation of the detected DEGs.
Project description:In the present study transcriptome analysis was employed to investigate the early molecular responses to exposure to the fungicide difenoconazole, a sterol biosynthesis inhibitor according to Fungicide Resistance Action Committee (FRAC) classification. Zebrafish embryos were exposed to difenoconazole according to OECD guidelines (OECD test No. 236). At the end of exposure time (96 hours), simultaneous RNA and protein extraction from 10 embryos was performed using a Macherey & Nagel RNA/protein extraction kit. The obtained RNA extracts were sequenced using Illumina HiSeq 4000 System and the obtained sequences went through bioinformatic analysis pipeline to Identify and count the detected gene sequences followed by differential gene expression analysis. Finally, potential substance specific biomarker candidates were refined and selected based on the differential expression patterns and the biological functions investigation of the detected DEGs.
Project description:Within the regulatory framework on the approval of new substances, the assessment of immunotoxic modes of action (MoA) is currently not covered. This is not least due to the lack of standardized methods and reliable validated biomarkers for immunotoxic effects. The experimental set up was designed to analyze the compound-specific response of zebrafish embryos to two immunomodulative reference substances, clobetasol propionate (CP, CAS 25122-46-7) and imiquimod (IMQ, CAS 99011-02-6), on the global level of gene expression. The obtained results were used to (i) identify potential biomarker candidates for the assessment of immunotoxic MoAs, (ii) identify compound-specific molecular signatures, (iii) evaluate the reliability of data acquisition using OMICs-coupled approaches and (iv) to assess the suitability of the zebrafish embryo as an alternative model for the human-representative investigation of psoriatic effects. For this, the transcriptomic profiles of embryos were bioinformatically analysed subsequent to an CP-induced immunosuppression in absence or presence of an IMQ-induced immune challenge, i.e. the simulation of a resting and an activated immune system. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to either CP (250 nM) or to IMQ (4000 nM) or to a combination of both under semi static conditions. Exposure to CP started at 2 hours post fertilization (hpf) until 72 hpf. Exposure to IMQ started at 48 hpf until 72 hpf. Untreated embryos reared in medium without CP nor IMQ dissolved was used as a negative control. All conditions comprised three biological replicates. Medium was exchanged on a daily basis by renewing half its volume. Occurrence of morphological changes and indicators of lethality as described in the OECD test guideline 236 were examined microscopically on a daily basis. At 72 hpf, all larvae were pooled for each sample for RNA extraction using a NucleoSpin RNA/Protein kit (Macherey-Nagel) following the manufacturer’s protocol. RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina NovaSeq 6000 System (Illumina) in 150 bp paired-end mode, producing a minimum of 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the Danio rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for differential gene expression analysis with DESeq2.
Project description:To systematically explore the functions of core splicing factors and regulators in alternative splicing, RNA-seq analyses were carried out upon the knockdown of over 305 splicing-related factors.
Project description:Endocrine active substances present significant risks to both human health and the environment, particularly by disrupting essential endocrine-regulated functions like organism development and reproductive capacity. Regulatory frameworks mandate animal testing for chemical risk assessment, placing specific emphasis on suspected endocrine disruptors. For example, aquatic vertebrate chronic toxicity testing is mandatory at a chemical tonnage of 100 t/y using the Fish Early Life Stage (FELS, OECD TG 210) test. However, suspected endocrine disruptors are subjected to testing from a lower tonnage threshold of 10 t/y. Currently, the assessment of endocrine effects adheres to an OECD Conceptual Framework (CF) that employs a tiered approach. This entails evaluating existing data for potential endocrine effects (Level 1), conducting mechanism of action-specific in vitro studies (Level 2), and undertaking in vivo mechanistic screening studies (Level 3). If screening studies indicate potential endocrine disruption, further investigation is conducted using a Fish Sexual Development Test (FSDT, OECD TG 234) to clarify (anti)estrogenic, (anti)androgenic, and steroidal effects (EAS effects) in fish. Nevertheless, this testing process is resource-intensive, time-consuming, and involves extensive animal use. Therefore, this study aims to identify the androgenic activity of trenbolone using the zebrafish embryo model. Trenbolone, a synthetic anabolic steroid, primarily exerts its effects by binding to androgen receptors. The data collected will be compared with those obtained from androstenedione exposure to zebrafish embryos.