Transcription profiling of mouse cerebella from pups exposed to PTU to induce hypothyroidism
Ontology highlight
ABSTRACT: Pregnant dams were treated with 0.1% or 0.04% 6-propyl-2-thiouracil (PTU) in drinking water continuously from day 13 post conception until weaning to produce hypothyroid pups. Cerebella were collected from vehicle and 0.1% PTU treated pups at post-natal day (PND) 15 and mRNA from these was subjected to microarray analysis using Agilent high-density oligonucleotide chips.
Project description:Pregnant dams were treated with 0.1% or 0.04% 6-propyl-2-thiouracil (PTU) in drinking water continuously from day 13 post conception until weaning to produce hypothyroid pups. livers were collected from vehicle and 0.1% PTU treated pups at post-natal day (PND) 15 and mRNA from these was subjected to microarray analysis using Agilent high-density oligonucleotide chips.
Project description:The use of custom-made focused microarrays has become a popular alternative, allowing the study of genes relevant to a specific hypothesis. To address the limitations of commercial arrays in toxicogenomics investigations, we have developed a custom mouse oligonucleotide microarray, the HC ToxArrayTM, which, by virtue of including genes that respond to a variety of chemical and physical stressors, is more relevant for toxicological studies. Furthermore, we have incorporated an extensive series of controls useful for both quality control and normalization of small focused arrays. Validation of the EC series for normalization was achieved by determining the gene expression profile in liver samples following treatment with the hepatotoxin phenobarbital. Technical variability experiments were also carried out by two different technicians using two hybridization methods (automated and manual) and compared. In this study, we demonstrate that dilution of oligonucleotides on the microarray itself provides an innovative approach allowing the full dynamic range of the scanner to be covered with a single gene spike-in. The dilution series can be used in a composite normalization to improve detection of differential gene expression and provide quality control measures.
Project description:A comparison of Ky mouse mutant soleus muscles versus wildtype soleus muscles. Hybridisation of 3 Mouse SGC 7.5k oligo slides, using 3 independent pools of 10 mice (5 male, 5 female)for both WT and mutant animals.
Project description:A comparison of Ostesy mutant soleus muscles to wildtype soleus muscles. Hybridisation of 3 Mouse SGC 7.5k oligo arrays, using 3 pools of 10 mice(5 male and 5 female) for mutant and WT.
Project description:An experiment was performed to understand its role in cell type specification, we have determined the human genomic binding sites of MLL1. MLL1 localizes with Pol II to the 5' end of actively transcribed genes, where histone H3 lysine 4 (H3-K4) trimethylation occurs. The ability of MLL1 to serve as a start site-specific global transcriptional regulator and to participate in larger chromatin domains at the Hox genes reveals the dual roles MLL1 plays in maintenance of cellular identity.
Project description:Dosage compensation ensures that males and females, despite unequal number of X chromosomes, equalize for X linked gene expression. In Drosophila, it is achieved by a two-fold up-regulation of most of the genes present on the male X chromosome, and requires the association of the Dosage Compensation Complex (DCC) on the X chromosome. One of the main intriguing aspects of dosage compensation is how this complex is able to target specifically hundreds of sites only on the X chromosome in order to ensure dosage compensation. In order to better understand the targeting of the DCC and the dosage compensation mechanism, we have then decided to analyze the distribution of the DCC as well as the expression levels in male and female in a more complete and precise manner, using microarrays. In this experiment, we present the data used to analyse the distribution of the MSL-1 and MSL-3 protein (part of the DCC complex) on the X chromosome in WT embryos aged from 0H-14H, as well as the distribution of MSL-1 in embryos aged from 4-6H and in male III instar salivary glands. The DNA amplified from the specific immunoprecipitation (MSL-1 or MSL-3 IP) were labelled using Cy5-dCTP and hybridized against DNA amplified from a non specific immunoprecipitation (mock IP), labeled with Cy3 dye. In parralel we present the data used to analyse the expression profile of X-linked genes in WT male or female III instar larvae salivary glands. The cDNA amplified from the RNA extracted from the male or female salivary glands were labelled using Cy5-dUTP and hybridized against the reference sample, labelled with Cy3-dUTP (pool RNA from ON embryos, adultes, and salivary glands mixed at a ratio 1:1:1.and amplified as the RNA from salivary glands). We used for this study a cDNA array developed by the Genecore facility in EMBL, covering the DGC1 and DGC2 cDNA libraries from the Berkeley Drosophila Genome Project, which represents more than 70% of the coding Drosophila genome.
Project description:Lactobacillus reuteri has been shown to encode a vitamin B12 biosynthesis pathway that is phylogenetically related to the one present in some representatives of gamma-proteobacteria such as Salmonella and Yersinia. Here we present evidence supporting that the similarities between these otherwise unrelated organisms extend to their regulatory mechanisms. Keywords: cell type comparison Loop design
Project description:This data set contains samples from Lactobacillus plantarum WCFS1 wild type and its CcpA mutant derivative. Both strains were grown in duplicate (samples L and R) and at four different growth stages samples were taken (early, mid and late log phase, early stationary phase). Samples were compared to each other within one strain and between the two strains. This allows the comparison of the wild type strain to its CcpA derivative in time. Further it allows comparison within one strain during different growth stages. Biological duplicate was performed.
Project description:In this study, we employ high-density oligonucleotide microarrays to characterize the MutaMouse FE1 cell line at various stages of cell growth, in primary MutaMouse lung epithelial cell cultures, and in whole lung. Global transcriptional analysis and real-time RT-PCR was applied to (1) further define the cellular origin of the FE1 cell line and its responses under different culture conditions (media and substratum), (2) provide insight into the transcriptional differences in cellular processes between FE1 cultures compared to whole lung tissues, more specifically in toxicological response, and (3) preliminarily examine FE1 culture response to exposure of benzo(a)pyrene compared to whole animals. Total RNA samples from 3 cell culture types (50% FE1, %100 FE1, and Primary lung) or MutaMouse lung were labeled with Cyanine 5-CTP, and universal reference total RNA (Stratagene, CA, USA) was labeled with Cyanine 3-CTP (Perkin Elmer Life Sciences, Woodbridge, ON, Canada) using Agilent linear Amplification kits (Agilent Tech. Inc. Mississauga, ON, Canada) following the manufacturer's instruction. Briefly, double-stranded cDNA was synthesized using MMLV-RT with T7 promoter primer, starting with 5 ug total RNA. Cyanine-labeled cRNA targets were in vitro transcribed using T7 RNA polymerase. The synthesized cRNA was precipitated by LiCl and fragmented at 60 degrees C for 30 min with fragmentation solution. Cy5- sample cRNA and Cy3- reference cRNA were hybridized to Agilent mouse development microarrays (containing ~20,000 unique 60 mer oligonucleotides; Agilent Tech. Inc. Mississauga, ON, Canada) at 60 degrees C overnight with Agilent hybridization solution and washed according to manufacturer's instruction. Arrays were scanned on a VersArray ChipReader (BioRad Laboratories Ltd., Waterloo, Ontario, Canada), and data were acquired with ImaGene 5.5 (BioDiscovery, Inc. CA, USA). Present calls were determined as signals that were greater than the mean plus three times the standard deviation of the average of the negative control spots.