Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of rat heart after treatment with PETN or NTG

ABSTRACT: Male Wistar rats (6 weeks, 250g), (Charles River, Sulzfeld, Germany) were equipped with micro osmotic pumps model 2001 from Alzet (Cupertino, CA) containing 400mM PETN dissolved in DMSO or the solvent and infusion was maintained for four days at 1?l/h (10.5?g/kg/min). For direct comparison, rats were also infused with 450mM NTG (6.6?g/kg/min in ethanol or the solvent as a control).
After 4d rats were sacrificed by exsanguinations under Isofluran anesthesia (5% inhalant in room air), the heart was rapidly excised, placed in ice-cold Krebs-HEPES-solution (composition in g/l: 5.78 NaCl, 0.35 KCl, 0.37 CaCl2, 0.30 MgSO4, 2.1 NaHCO3, 0.14 K2HPO4, 5.21 HEPES and 2.0 D-glucose) and dissected.
Total RNA was isolated from the hearts. Direct labeled cDNA was synthezised and hybridized to Rat OpArray Microarray (total genom expression profiling).

ORGANISM(S): Rattus norvegicus

SUBMITTER: Hartmut Kleinert

PROVIDER: E-MEXP-2043 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Similar Datasets

Project description:Bone marrow (BM) cells derived from femurs of C57BL/6 mice were used to generate BM-DCs. For this, BM cells (2x106 cells/10 ml) were seeded on 100 mm2 bacterial dishes (Greiner bio-one, Frickenhausen, Germany). DC culture media (IMDM, 5% [v/v] FCS [PAA, Coelbe, Germany], 100 U/ml penicillin, 100 ug/ml streptomycin [both Gibco, Paisly, UK], 50 uM beta-mercaptoethanol [Roth, Karlsruhe, Germany] 5% [v/v] of murine GM-CSF containing cell culture supernatant was replenished on days 3 and 6 of culture. Aliquots of DC cultures were treated with recombinant human 100 ng/ml WNT-1 (Biovision, Milpitas, CA) for 48h on days 7 or 8 of culture. Each 20 ug of total RNA derived from unstimulated BM-DCs treated for 48 h with WNT-1 (100 ng/ml) or left untreated was used for generation of fluorescecne-labeled cDNA probes (Alexa Fluor 555-aha-dUTP for untreated BM-DCs; Alexa Fluor 647-aha-dUTP for WNT-treated BM-DCs). <br><br><br><br>Prehybridized (5X SSPE,0.1% SDS, and 1% BSA) microarrays (Mouse Whole Genome OneArray Microarray v2; Phalanx Biotech, San Diego, U.S.A.) were hybridized with each 20 pmol of either fluorescence-labeled cDNA probe using OneArray Hybridization Buffer with 18% formamide for 16 h (Lucidea SlidePro Hybridizer; GE Healthcare, Munich, Germany) according to the OneArray hybridization protocol. Washed microarrays were analysed using a ScanArray 4000 system equipped with ScanArray 4.0 software (both Perkin-Elmer; Rodgau-Jugesheim, Germany) at 10 pixel size. Laser intensity (90-100%) and photomultiplier tube sensitivity (60-70%) were varied to optimize signal to background ratios. Derived images weere analyzed using the ScanArray 4.0 easyquant software module. Intensity data were normalized (loess normalization; MIDAS-2.19) and subjected to significance analysis (MEV 4.3) using appropriate modules of the TM4 microarray software suite. Changes in gene expression with p < 5% were used for subsequent analyses. Gene ontology annotations were performed with Panther software (Thomas et al., 2003).

Project description:The two-component system (TCS) is a specific regulatory system in bacteria and plays an important role in sensing and adapting to the environment. In this study, we evaluated the roles of TCSs in the major cariogenic pathogen Streptococcus mutans in resistance to several types of bacteriocin. In a comprehensive analysis using individual TCS mutants, we found that two novel TCSs were associated with resistance against distinct lantibiotics nisin A (class I type A[I]) and nukacin ISK-1(class I type A[II]). One TCS, SMU.659-660 (designated as NsrRS), was related to resistance against nisin A produced by Lactococcus lactis ATCC 11454. The other TCS, SMU.1146-1145 (designated as LcrRS), was related to resistance against nukacin ISK-1 produced by Staphylococcus warneri ISK-1. NsrRS induced the expression of SMU.658 (designated as NsrX), which constitutes an operon with nsrRS, in response to nisin A. Inactivation of nsrX increased susceptibility to nisin A. Additionally, NsrX expression in S. mutans increased the binding affinity to nisin A compared to a no-expression strain. LcrRS induced the expression of SMU.1148-50 (lctFEG), which encodes an ABC transporter and is located upstream of lcrRS, in response to nukacin ISK-1. Inactivation of lctFEG significantly increased susceptibility to nukacin ISK-1. Electrophoretic mobility shift assay analysis revealed that NsrR and LcrR bound directly to regions upstream of nsrX and lctFEG, respectively. This is the first report that two distinct TCSs, NsrRS and LcrRS, are independently involved in resistance to nisin A and nukacin ISK-1 in S. mutans. Total of 14 samples were analyzed. Total RNA from each test strain and control described below were labeled with Alexa FluorÂ® 555 and Alexa FluorÂ® 647, respectively, and were cohybridized on a single array. Labeling and hybridization were performed once or twice independently. UA159 wild type as control vs. UA159 with nisinA or nukacin ISK-1, UA159 wild type with nisinA or nukacin ISK-1 vs. nsrRS or lcrRS deletion mutant in UA159 with nisinA or nukacin ISK-1.

Dataset Information

Transcription profiling of rat heart after treatment with PETN or NTG

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets