Dataset Information

Early embryonic gene expression profiling of zebrafish prion protein (Prp2) morphants.

ABSTRACT: BACKGROUND: The Prion protein (PRNP/Prp) plays a crucial role in transmissible spongiform encephalopathies (TSEs) like Creutzfeldt-Jakob disease (CJD), scrapie and mad cow disease. Notwithstanding the importance in human and animal disease, fundamental aspects of PRNP/Prp function and transmission remains unaccounted for. METHODOLOGY/PRINCIPAL FINDINGS: The zebrafish (Danio rerio) genome contains three Prp encoding genes assigned prp1, prp2 and prp3. Currently, the second paralogue is believed to be the most similar to the mammalian PRNP gene in structure and function. Functional studies of the PRNP gene ortholog was addressed by prp2 morpholino (MO) knockdown experiments. Investigation of Prp2 depleted embryos revealed high mortality and apoptosis at 24 hours post fertilization (hpf) as well as impaired brain and neuronal development. In order to elucidate the underlying mechanisms, a genome-wide transcriptome analysis was carried out in viable 24 hpf morphants. The resulting changes in gene expression profiles revealed 249 differently expressed genes linked to biological processes like cell death, neurogenesis and embryonic development. CONCLUSIONS/SIGNIFICANCE: The current study contributes to the understanding of basic Prp functions and demonstrates that the zebrafish is an excellent model to address the role of Prp in vertebrates. The gene knockdown of prp2 indicates an essential biological function for the zebrafish ortholog with a morphant phenotype that suggests a neurodegenerative action and gene expression effects which are apoptosis related and effects gene networks controlling neurogenesis and embryo development.

SUBMITTER: Nourizadeh-Lillabadi R

PROVIDER: S-EPMC2962645 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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Publications

Early embryonic gene expression profiling of zebrafish prion protein (Prp2) morphants.

Nourizadeh-Lillabadi Rasoul R Seilø Torgersen Jacob J Vestrheim Olav O König Melanie M Aleström Peter P Syed Mohasina M

PloS one 20101022 10

<h4>Background</h4>The Prion protein (PRNP/Prp) plays a crucial role in transmissible spongiform encephalopathies (TSEs) like Creutzfeldt-Jakob disease (CJD), scrapie and mad cow disease. Notwithstanding the importance in human and animal disease, fundamental aspects of PRNP/Prp function and transmission remains unaccounted for.<h4>Methodology/principal findings</h4>The zebrafish (Danio rerio) genome contains three Prp encoding genes assigned prp1, prp2 and prp3. Currently, the second paralogue ...[more]

PMID: 21042590

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Project description:Sensitivity and throughput of transcriptomics and proteomomics technologies have advanced tremendously in recent years. With the use of deep sequencing of RNA samples (RNAseq) and mass spectrometry technology for protein identification and quantitation, it is now feasible to compare gene and protein expression on a massive scale and on any organism for which genomic data is available. Although these technologies are currently applied to many research questions in various model systems ranging from cell cultures to the entire organism level, there are few comparative studies of these technologies in the same system, let alone on the same samples. Here we present a comparison between gene and protein expression in zebrafish embryos, which is an upcoming model in disease studies. We compared Agilent custom made expression microarrays with Illumina deep sequencing for RNA analysis, showing as expected a high degree of correlation of expression of a common set of 15,927 genes. Gene expression was also found to correlate with the abundance of 1,017 quantified proteins, with several categories of genes as exceptions. These exceptions include ribosomal proteins, histones and vitellogenins, for which biological and technical explanations are discussed. By comparing state of the art transcriptomic and proteomic technologies on samples derived from the same group of organisms we have for the first time benchmarked the differences in these technologies with regard to sensitivity and bias towards detection of particular gene categories. Our datasets submitted to public repositories are a good starting point for researchers interested in disease progression in zebrafish at a stage of development highly suited for high throughput screening technologies. The transcriptomics data is deposited in NCBI GEO. Proteomics informatics: Spectra were identified by Mascot and validated by PeptideProphet and ProteinProphet in the Trans-Proteomic Pipeline (TPP). The main search parameters are: full tryptic specificity, precursor mass searched within -0.5 to +2.5 Da, product ions within [-0.5,0.5] Da and allowing for one missed cleavage. Carbamidomethylation was assumed as the only and fixed PTM.

Dataset Information

Early embryonic gene expression profiling of zebrafish prion protein (Prp2) morphants.

Publications

Early embryonic gene expression profiling of zebrafish prion protein (Prp2) morphants.

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