Project description:We generated a 16 nucleotide deletion mutant in the zebrafish fbln1 gene and we compared the whole larvae transcriptome to wt siblings in 10dpf larvae.
Project description:Wild type zebrafish (AB strain, https://zfin.org/action/genotype/view/ZDB-GENO-960809-7) were set up for mating overnight, in 3:2 ratios, females and males, respectively. Fertilized embryos from wildtype zebrafish AB strain were used for all experiments and kept in Embryo medium (E3, 5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2, 0.33 mM MgSO4) in an incubator at 28ºC. At 1dpf, larvae were manually dechorionated under a stereomicroscope. At 2dpf, zebrafish larvae were divided randomly into the different experimental groups, at a density of 12 larvae per well of a 12 well plate (Corning) in 1.5mL of E3. Three biological replicates of 12 larvae per condition were exposed to different conditions for 4 hours at 28 ºC: DMSO; DMSO + CH223191 (5mM); 3-o-C12-L-HSL (50mM); 3-o-C12-L-HSL (50mM) + CH223191 (5 mM ); 1-hydroxyphenazine (1-HP, 5mM); 1-HP (5mM) + CH223191(5mM) and 1-HP (5mM) + 3-o-C12-L-HSL (50mM).To mention, in experiments using the Aryl hydrocarbon Receptor inhibitor CH223191, larvae were pre-exposed to 5uM of the inhibitor for 2h prior to the start of the experiment and the inhibitor was kept during the experiment. After the desired exposure to diverse ligands, larvae were euthanized with Tricaine (MS-222, 300 µg/mL) and placed in Trizol for RNA isolation. The same experimental layout was performed 5 times, and samples were subjected to microarray hybridization and analysis.
Project description:The zebrafish Cxcr3.2 is a functional homolog of the human chemokine receptor CXCR3. Zebrafish macrophages lacking this receptor have impaired motility and a rounded shape compared to their wildtype counterparts. To investigate the effects of cxcr3.2 mutation on the transcriptional profile of macrophages, we sorted macrophages from zebrafish larvae lacking a functional cxcr3.2 and compared their transcriptome to that of macrophages from wildtype larvae. Mutant and wildtype macrophages could be clearly distinguished based on the overall differential expression profiles. Classification of genes by compartment showed that peroxisomal, lysosomal and Golgi-related genes were most frequently up-regulated. Moreover, lysosomal and Golgi-related terms were significantly differentially represented in Gene Ontology and KEGG enrichment analysis. Of note, several lysosomal markers (including acidic hydrolases and voltage ATPases) were consistently upregulated in cxcr3.2 mutant macrophages, indicating that cxcr3.2-mediated chemokine signaling is tightly connected to the regulation of lysosomal function.
Project description:Transcriptional profiling of 3dpf wild type zebrafish larvae treated with 20mM PTZ for 30 and 90 minutes compared with 3dpf wild type control untreated zebrafish larvae.
Project description:DNA methylation is a central repressive epigenetic mark. In this dataset we performed RRBS on whole zebrafish larvae from uhrf1-hi272 mutants and phenotypically wild-type siblings to determine which genomic regions have changes in DNA methylation due to uhrf1 loss.
Project description:To understand the effects of Hsp60 deficiency in developing vertebrates, we generated CRISPR/Cas9-mediated hspd1 knockout zebrafish lines by targeting exon 2 to induce a frameshift mutation. We selected an allele with a 56 base pair deletion inducing a frameshift mutation leading to loss of protein functions. We examined the proteome changes in zebrafish larvae at 5 days post fertilization (DPF). Wildtype control and hspd1-/- larvae at 5dpf, were analyzed by TMT and nanoLC-MS/MS based proteomcis. For this purpose, we studied five pools from each genotype, and each pool consisted of five larvae.
Project description:In this work we investigated how the brain proteome of the larval zebrafish is modified by behavioral adaptation to the environmental challenge of a water vortex. We monitored the behavior of larvae and observed that they behaviorally adapted to the presence of a water vortex. We obtained the larval zebrafish brain proteome by extracting brains from zebrafish larvae and analyzing them using and LFQ-based LC-MS/MS-approach. In total we identified 5929 proteins in the larval brain. Within this proteome, we identified 57 proteins that were significantly regulated following experience of the water vortex: 41 proteins were up regulated and 16 were down regulated. Of these, 29 proteins are known to have neuronal functions, 17 proteins are known to have other cellular functions, and 11 proteins are still uncharacterized.