ABSTRACT: The aim of this study was to identify genes that are regulated by ccm2 signaling with the zebrafish heart at around 72 hours post fertilization. In total, 4 samples based on cDNA from extracted cardiac tissue were analyzed: 2 samples of ccm2 m201 mutant embryos and 2 WT control samples.
Project description:The aim of this study was to identify genes that are regulated by Bmp signaling with the zebrafish heart at the heart cone stage around 21-23 hours post fertilization. In total, 4 samples based on cDNA from extracted cardiac tissue were analyzed: 2 samples of Tg(hsp70l:bmp2b)fr13 transgenic embryos heatshocked at 18 hpf and 2 WT control samples.
Project description:Key tissues of the adult female parasite involved in nutritional uptake and reproduction were examined using a novel gene discovery approach that combined laser microdissection microscopy and microarray analyses. Gastrodermis, vitelline glands and ovary were microdissected from unfixed, frozen sections of the Asian species, Schistosoma japonicum. Total RNA was isolated from the enriched tissue preparations and microarray analyses undertaken to generate tissue specific gene expression profiles. Single colour adult female schistosome laser microdissection microarray data with technical replicates in duplicate: Gastrodermis (gut); vitelline glands (vitelline); ovary; and whole section (control).
Project description:To define the differential gene regulation of adult Schistosoma japonicum in response to host insulin, we utilised a parasite microarray with 38,444 probes representing 19,222 genes. We cultured male and female adult worms were exposed to insulin and glucose; and then parasites were collected at three time-points. There were 1,437 probes (1101 genes) up or down regulated (±1.5 fold change) by insulin after 0.5, 3, 24 hours, with the majority, 712 probes (577 genes) and 623 probes (486 genes) were up-regulated at 24 hour of adding insulin in males and females respectively compared with the control (without insulin). Single colour microarray, insulin treated adult Schistosoma japonicum with technical replicates in duplicate: Separate male and female worms exposed to insulin at 0, 0.5, 3, 24 hours. This study includes 6 control samples for separate male and female worms cultured without insulin at 0, 0.5, 3, 24 hours.
Project description:We have examined the gene expression profile of the transformation of the blood fluke Schistosoma mansoni from the cercarial to schistosomula stages. Specifically cercariae were mechanically transformed and then cultured for 3 hours or 5 days, and then total RNA was isolated and microarray analysis performed. A second experiment compares the effect that the presence of red blood cell in the culturing conditions has over the 3 hour and 5 day periods have on the parasite. Agilent custom designed microarray was used. One colour experimental design. One biological replicate was labelled and then two technical replicates (hybridisations) performed for each sample. A total of 5 samples were examined, cercariae, 3 hrs +RBCs, 5 days + RBCs, 3 hrs â??RBCs and 5 days â?? RBCs.
Project description:The zebrafish (Danio rerio) is a popular animal model in studies of vertebrate development and organogenesis. Recent research has shown a similarity of approximately 70% between the human and zebrafish genomes and of 84% in human disease-causing genes, specifically. Zebrafish embryos have a number of desirable features, including transparency, a large size, and rapid embryogenesis. Protein phosphorylation is a well-known post-translational modification (PTM), which performs various biological functions. Recent mass spectrometry (MS) developments have enabled the study of global phosphorylation patterns by using MS-based proteomics coupled with TiO2 phosphopeptide enrichment. In the present study, we identified 3,500 non-redundant phosphorylation sites on 2,166 phosphoproteins and 1,564 quantified phosphoproteins in zebrafish embryos.
Project description:Heart formation requires input from two populations of progenitor cells - the first and second heart fields - that differentiate at distinct times and create different cardiac components. The cardiac outflow tract (OFT) is built through recruitment of late-differentiating, second heart field (SHF) -derived cardiomyocytes to the arterial pole of the heart. Mechanisms responsible for selection of an appropriate number of OFT cells from the SHF remain unclear, although several lines of evidence emphasize the importance of FGF signaling in promoting this process. Here, we examine the impact of inhibition of FGF signaling on cardiac transcription profiles in an effort to identify genes operating downstream of FGF during OFT development. We compared hearts from embryos treated with the FGFR inhibitor SU5402 to the hearts from sibling embryos treated with DMSO. Two replicates were performed.
Project description:We identified a novel paralog of the antioxidant response element transcription factor, Nrf2 in zebrafish. To determine whether the paralogs Nrf2a and Nrf2b regulate different sets of genes, we performed loss-of-function experiments followed by microarray-based gene expression profiling, performed on embryos in which expression of Nrf2a, Nrf2b, or both paralogs was knocked down. Embryos at the 1-4 cell stage were injected with 3-5 nl of a control morpholino (MO) or gene-specific MO to knockdown Nrf2a, Nrf2b, or a Nrf2a+b combination. At 48 hpf, triplicate groups of 5 embryos were exposed to 2 M-BM-5M of tert butyl hydroquinone or 2% DSMO for 4 hours. At 52 hpf, embryos were fixed in RNA Later and stored at -80M-BM-0C. RNA was extracted, bioanalyzed, labeled with Cy3 and hybridized to the Agilent V3 4x44K zebrafish microarray (cat. #G2519F-026437) at the Genome Technology Core of the Whitehead Institute (Cambridge, MA). Raw array data obtained from the Whitehead Institute were extracted using Agilent's feature extraction software using background detrending (spatial and multiplicative).Prior to normalization, Cy3 values below 5 were set to 5. The data were then normalized using the non-linear scaling method based on rank invariant probes. After normalization but before statistical analyses, probes not significantly above background in all microarrays were removed (3147 probes in all; based on AgilentM-bM-^@M-^Ys 2.6 standard deviation method). None of the probes were saturated for Cy3 signal on any microarray, so no further filtering was applied. There were a total of 40,456 probes for statistical analyses. Statistical tests were performed using MeV v4. Data were log transformed and median centered for each probe. An ANOVA was run for morpholino injection, with p-value based on 1000 permutations of the data and alpha of 0.01. The probes found significant in the ANOVA were subsequently examined using Rank Product (RP) analysis to identify probes up- and down-regulated by the MO injection For each RP test, a two-class unpaired RP analysis was performed using 100 permutations of the data with a false discovery rate (FDR) M-bM-^IM-$10%.
Project description:Investigation of whole genome expression pattern of 60 and 72 hours post fertilization Danio Rerio embryos exposed to TMT and vehicle control Embryos were exposed to 10uM TMT or control from 48hpf to 60 or 72 hpf. Three replicates were collected for each time point. 40 embryos were pooled to comprise a replicate.
Project description:Doxorubicin (DOX) is an effective anthracycline agent used to combat many neoplastic diseases. However, DOX causes cardiovascular toxicity in juvenile and young adult cancer survivors that can lead to future cardiomyopathy. Thus, it is essential to address the cardiovascular toxicity caused by DOX to improve the long-term health of cancer patients. Several studies have suggested that soluble epoxide hydrolase (sEH) and cyclooxygenase-2 (COX-2) are implicated in cardiovascular diseases by impairing vascular health and promoting the transition of Endothelial cells to Mesenchymal cells (EndMT). Given the role of sEH and COX-2 in EndMT cardiovascular toxicity, we aimed to investigate the effect of a dual sEH/COX-2 inhibitor, PTUPB, on DOX-induced EndMT, vascular and cardiac toxicity. We tested the beneficial effect of PTUPB on DOX-induced cardiovascular toxicity in zebrafish.
Project description:In the present study, a oligonucleotide microarray platform is used to compare expression profiles of beef cattle muscle in animals treated with either Dexamethazone (Dex) or Dexamethazone plus 17?-estradiol (Estr) administered at sub-therapeutic dosage, against untreated controls. Seventeen male beef cattle 15-18 months old, around 450 kg mean body weight were randomly allocated in three groups: 6 were untreated (group Ctrl), 5 were administered with dexamethazone via feed 0.75mg/head for 43 days (group D); the last 6 animals were administered via feed for 43 days with Dex (0.75mg/head) and intramuscularly (i.m.) for three times with 17?-oestradiol, 20mg/head, (group DE). Three additional control animals, matching in sex and age, collected in a previous experiment were included in the Ctrl group to increase sample size and to control for biological variation. For each sample, total RNA was extracted from a specific anterior limb muscle (biceps branchii). Data analysis demonstrates that the expression profiles were strongly affected by Dex treatment with hundreds of genes up-regulated with relevant fold-change, whereas the myostatin gene was significantly down-regulated. On the contrary, the administration of Dex-Estr reveals a weaker effect on gene expression. In this study, we analyzed the gene expression profiles of 20 anterior limb muscle samples, 9 collected from untreated controls, 5 collected from cattle treated with Dex and 6 from cattle treated with Dex+Estr using Agilent-015354 Bovine oligo microarray platform (20 arrays, no replicate) based on single-colour detection (Cyanine-3 only). Microarrays are scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides are scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software creates a unique ID for each pair of XDR scans and saves it to both scan image files. Feature Extraction 9.5 uses XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal