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Metabolic Profile Analysis of Zebrafish Embryos
Published on: January 14, 2013
A dataset of transcriptomic effects of camptothecin treatment on early zebrafish embryos
Sergey V Prykhozhij1, Kevin Ban1, Zane L Brown2
1Children's Hospital of Eastern Ontario (CHEO) Research Institute and University of Ottawa, Ottawa, ON, Canada.
Abstract:
Zebrafish (Danio rerio) are a good model for cancer research including studies on chemotherapy treatments. We treated wild-type and miR-34a deletion mutant zebrafish embryos at 24 h post-fertilization with 1 µM of the topoisomerase I inhibitor, camptothecin (CPT), for 4 h to catalogue gene expression changes induced by this DNA damage treatment and to understand if these changes are influenced by loss of miR-34a. The 4 sample groups of 3 independent biological samples consisting of 30 embryos each were analyzed by RNA-sequencing using the recently updated zebrafish transcriptome annotation based on GRCz11, which enabled a more complete and sensitive read mapping and gene assignment than standard annotations. Using this gene expression estimates dataset as the primary resource, we performed a differentially expressed gene (DEG) analysis based on treatment as loss of miR-34a had minimal effects on CPT-induced expression changes. The DEGs were analyzed for Gene Ontology and KEGG pathway terms. Enriched terms and pathways among up-regulated genes were mostly related to stress, cell death, cell cycle regulation, transcriptional regulation, cell signalling, developmental processes and synthesis of retinol and steroid hormones. By contrast, down-regulated genes were most strongly associated with genes involved in key developmental processes, adhesion molecules, as well as some transport and metabolic pathways, together suggesting a "developmental shutdown". We also identified interferon-regulated genes and p53 target genes activated or inhibited by DNA damage due to topoisomerase I inhibition, suggesting that they are important components of the response to this type of DNA damage in zebrafish embryos.
Insights
Zebrafish embryos treated with camptothecin show significant gene expression changes related to stress and development. Loss of miR-34a had minimal impact on these chemotherapy-induced DNA damage responses.
Area of Science:
- Developmental Biology
- Genomics
- Cancer Research
Background:
- Zebrafish (Danio rerio) serve as a valuable model organism for studying cancer and chemotherapy treatments.
- MicroRNA-34a (miR-34a) plays a role in cellular processes, and its function in response to DNA damage is under investigation.
Purpose of the Study:
- To catalogue gene expression changes in zebrafish embryos induced by camptothecin (CPT), a topoisomerase I inhibitor.
- To investigate the influence of miR-34a deletion on CPT-induced gene expression alterations.
Main Methods:
- Wild-type and miR-34a deletion mutant zebrafish embryos were treated with CPT.
- RNA-sequencing was performed using an updated zebrafish transcriptome annotation (GRCz11).
- Differentially expressed gene (DEG) analysis and pathway enrichment (Gene Ontology, KEGG) were conducted.
Main Results:
- CPT treatment induced significant gene expression changes, primarily related to stress, cell death, cell cycle regulation, and developmental processes.
- Down-regulated genes suggested a "developmental shutdown" response.
- Loss of miR-34a had minimal effect on CPT-induced expression changes.
- Interferon-regulated and p53 target genes were identified as key components of the DNA damage response.
Conclusions:
- Zebrafish embryos exhibit a complex gene expression response to topoisomerase I inhibition, involving stress and developmental pathways.
- miR-34a appears to have a limited role in modulating the immediate transcriptional response to CPT-induced DNA damage in this model.
- The study highlights the utility of updated genomic annotations for sensitive gene expression analysis in zebrafish.

