The impact of exposure to serum lipids during in vitro culture on the transcriptome of bovine blastocysts
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ABSTRACT: EtOH-Lipid form serum have been added in BSA-supplemented SOF during post-compaction development of IVP bovine embryos. Lipid-treated blastocysts were compared with control treated blastocysts (no lipid addition to BSA, EtOH balanced). Amplified anti-sense RNA from each sample (4 pools of control and 4 pools of treated blastocysts) were labeled then hybridized on oligonucleotide microarrays following a dye-swap design (ex: pool 1 ctl in red vs. pool 1 treatment in green and then pool 1 ctl in green vs. pool 1 treatment in red).
Project description:IVP blastocysts exposed to hyperglycaemia during early phase of development, i.e from zygote to 8-16 cells stages. Genes expression analysis is done at day 7.5 (7 days of embryo culture). Total RNA form glucose treated blastocysts (4 replicates of 10 embryos each) and control treated blastocysts (4 replicates of 10 embryos each) were compared on a 2-color micro-array design and dye swap.
Project description:Bovine cloned and IVF blastocysts at day 7 were collected and used for comparative transcriptomics. A potent histone deacetylase inhibitor, Trichostatin A (TSA), was used to understand the effect of assisted epigenetic modification on the transcriptional profile of SCNT blastocysts and to identify specific genes or categories of genes affected in the developing embryos.
Project description:The deubiquitinase BAP1 is a candidate tumor suppressor regulating cell proliferation in human and is required for development in Drosophila. BAP1 is assembled into high molecular weight transcriptional multi-protein complexes. In order to identify potential BAP1 target genes, global mRNA expression profiling using microarrays was conducted. U2OS cells, transfected with a non-target control shRNA or shRNAs targeting BAP1, were selected with puromycin containing medium and then synchronized at the G1/S border to allow comparative analysis of gene expression.
Project description:Cinnamaldehyde is a natural antimicrobial and has been found to be effective against many foodborne pathogens including Escherichia coli O157:H7. Although its antimicrobial effects have been well investigated, limited information is available on its effects at the molecular level. Sublethal treatment at 200 mg/l cinnamaldehyde inhibited growth of E. coli O157:H7 at 37oC and for M-bM-^IM-$ 2 h caused cell elongation, but from 2 to 4 h growth resumed and cells reverted to normal length. To understand this transient behaviour, genome-wide transcriptional analysis of E. coli O157:H7 was performed at 2 and 4 h exposure to cinnamaldehyde. Drastically different gene expression profiles were obtained at 2 and 4 h. At 2 h exposure, cinnamaldehyde induced overexpression of many oxidative stress-related genes, reduced DNA replication, and synthesis of protein, O-antigen and fimbriae. At 4 h, many cinnamaldehyde-induced repressive effects on E. coli O157:H7 gene expressions were reversed and oxidatve stress genes were nolonger differentially expressed. Duplicate E. coli O157:H7 cultures with or without 200 mg/l cinnamaldehyde were incubated at 37M-BM-0C for M-bM-^IM-$ 4 h. Cinnamaldehyde-induced changes in gene expression profiles were compared at 2 and 4 h using Affymetrix Ginechip 2.0 microarrays.
Project description:This SuperSeries is composed of the following subset Series: GSE42279: AAPH-treated blastocysts vs CTL-treated blastocysts GSE42280: BSO-treated blastocysts vs CTL-treated blastocysts Refer to individual Series
Project description:Microarray analysis of transcriptome bovine blastocysts Day 8, deriving from oocytes matured under different insulin concentrations as model for metabolic imbalance Three conditions experiment: control, Low Insulin, High Insulin treatments. Biological replicates: 4 for each treatment. One replicate per array.
Project description:Lipo-chitooligosaccharides (LCOs) produced by N2-fixing rhizobacteria initiate host nodule formation. Foliar application of LCOs has been shown to induce stress-related genes under optimal growth conditions. To study the effects of LCO foliar spray under stressed conditions, soybean seedlings grown at optimal temperature were exposed to sub-optimal temperature. After a 5-day acclimation period, the first trifoliolate leaves were sprayed with 10-7 M LCO (NodBj-V (C18:1, MeFuc)) produced by Bradyrhizobium japonicum, and harvested at 0 and 48 h following treatment. Microarray analysis was performed using Affymetrix GeneChip® Soybean Genome Arrays. A total of 147 genes were differentially expressed 48 h after LCO treatment, including a number of stress-related genes and transcription factors. In addition, during the 48 h following treatment, hundreds of genes were differentially expressed in LCO-treated plants, indicating that the dynamic soybean foliar transcriptome was highly responsive to LCO treatment. The microarray data was supported by quantitative real-time PCR data. Soybean seedlings grown at optimal temperature (25 °C) were exposed to sub-optimal temperature (15 °C). After a 5-day acclimation period, the first trifoliolate leaves were sprayed with 10-7 M LCO (NodBj-V (C18:1, MeFuc)) produced by Bradyrhizobium japonicum, and harvested at 0 and 48 h following treatment. Total RNA was extracted and microarray analysis was performed using Affymetrix GeneChip® Soybean Genome Arrays.
Project description:In order to understand how in vitro culture affects embryonic quality, we analyzed survival and global gene expression in bovine blastocysts after exposure to increased oxidative stress conditions of IVC. A pro-oxidant agents that act intra-cellularly by inhibiting GSH synthesis (0.4 mM buthionine sulfoximine [BSO]) was added from days 3 to 7, and transcriptomic analysis was then performed in resulting blastocysts. Precisely, after in vitro maturation and fertilization, bovine zygotes were culture in vitro in normal condition, then at day 3, embryos were allocated into culture in control or supplemented with BSO (0.4 mM) until day 7. At this time, blastocysts were harvested and analyzed. Our hypothesis was that BSO treatment will affect blastocyst survival and gene expression associated with low embryo quality 4 replicates of 10 blast/rep was produced for BSO and control treatment. According to a dye swap design with 2 colors, 4 arrays were used to compare BSO (green) against CTL (red) and 4 arrays were used to compared BSO (red) againt CTL (green).
Project description:In order to understand how in vitro culture affects embryonic quality, we analyzed survival and global gene expression in bovine blastocysts after exposure to increased oxidative stress conditions of IVC. A pro-oxidant agents that act extra-cellularly by promoting ROS production (0.01 mM 2,2'-azobis (2-amidinopropane) dihydrochloride [AAPH]) was added from days 3 to 7, and transcriptomic analysis was then performed in resulting blastocysts. Precisely, after in vitro maturation and fertilization, bovine zygotes were culture in vitro in normal condition, then at day 3, embryos were allocated into culture in control or supplemented with AAPH (0.01 mM) until day 7. At this time, blastocysts were harvested and analyzed. Our hypothesis was that AAPH treatment will affect blastocyst survival and gene expression associated with low embryo quality 4 replicates of 10 blast/rep was produced for AAPH and control treatment. According to a dye swap design with 2 colors, 4 arrays were used to compare AAPH (green) against CTL (red) and 4 arrays were used to compared AAPH (red) againt CTL (green).
Project description:Transcriptional profiling of porcine expanded blastocysts comparing control (EB obtained from 4 sows treated with basal diet) with either inorganic Se + B6 (EB obtained from 4 sows treated with basal diet plus inorganic Se and B6) or organic Se + B6 (EB obtained from 3 sows treated with basal diet plus organic Se and B6). Three-condition experiment, EB without and with maternal diet supplemented B6 plus either inorganic Se or organic Se. Four biological replicates for inorganic Se and three biological replicates and one technical replicate for organic Se. Pooled of four biological replicates for control group.