Project description:This SuperSeries is composed of the following subset Series: GSE28410: Mouse oocytes: High hydrostatic pressure (HP) treated vs. Control GSE28411: Mouse in vitro fertilized four-cell stage embryos: High hydrostatic pressure (HP) treated vs. Control Refer to individual Series
Project description:Mouse oocytes were treated with 20 MPa hydrostatic pressure for 60 minutes at 37 °C. The HHP treated oocytes and their untreated controls were fertilized by ICSI and cultured till four-cell stage, when the transcription profiling experiment was performed. One-condition experiment, Four-cell stage embryos from HP treated oocytes vs. four-cell stage embryos from control oocytes. Biological replicates: 3 HP treated replicates, 3 control replicates.
Project description:Mouse oocytes were treated with 20 MPa hydrostatic pressure for 60 minutes at 37 °C. The HHP treated oocytes and their untreated controls were fertilized by ICSI and cultured till four-cell stage, when the transcription profiling experiment was performed.
Project description:Transcription profiling of mouse oocytes treated with 20 MPa hydrostatic pressure for 60 minutes at 37 °C comparing control oocytes kept under identical conditions as pressure treated ones, except HHP treatment. One-condition experiment, HP treated oocytes vs. Control oocytes. Biological replicates: 4 HP treated replicates, 4 control replicates.
Project description:Gene expression in human optic nerve head (ONH) astrocytes exposed to either 60 mm Hg hydrostatic pressure (HP) or control ambient pressure (CP) was compared using Affymetrix GeneChip microarrays to identify HP-responsive genes. Primary ONH astrocytes from two male Caucasian donors (passage 4) were grown to 75% confluence and were exposed for 6, 24 or 48 h to control ambient pressure (CP6, CP24, CP48) or hydrostatic pressure (HP6, HP24, HP48), or harvested at the beginning of the pressure experiment (CP0). Total RNA was extracted using Qiagen RNeasy columns and converted to biotin-labeled cRNA by standard Affymetrix protocols available at web site http://pathology.wustl.edu/~mgacore/genechip.htm#Preparing. Hybridization of the labeled cRNA to Human Genome U95Av2 chips (Affymetrix) was carried out by using Genechip Instrument System (Affymetrix) at Genechip Core Facility of Washington University. A total of 44 chips were generated and distributed as follows: seven for control pressure (CP) at time 0, four CP at 6 h, seven for hydrostatic pressure (HP) at 6 h, eight for CP at 24 h, eight for HP at 24 h, five for CP at 48 h and five for HP at 48h P. The arrays were washed and stained with streptavidin-phycoerythrin followed by scanning on an Agilent GeneArray Scanner G2500A (Agilent Technologies, Palo Alto, CA), and then scanned by an Affymetrix GeneArray Scanner. Data was analyzed by Affymetrix Microarray Suite (version 5.0), linear regression analysis, and GeneSpring expression Analysis Software (version 6.0, Silicon Genetics). For analysis, the samples were scaled to the same target intensity (1500) to allow comparison of multiple samples, and raw data were normalized to median of each gene across all chips for fold change analysis using GeneSpring.
Project description:Gene expression in human optic nerve head (ONH) astrocytes exposed to either 60 mm Hg hydrostatic pressure (HP) or control ambient pressure (CP) was compared using Affymetrix GeneChip microarrays to identify HP-responsive genes. Primary ONH astrocytes from two male Caucasian donors (passage 4) were grown to 75% confluence and were exposed for 6, 24 or 48 h to control ambient pressure (CP6, CP24, CP48) or hydrostatic pressure (HP6, HP24, HP48), or harvested at the beginning of the pressure experiment (CP0). Total RNA was extracted using Qiagen RNeasy columns and converted to biotin-labeled cRNA by standard Affymetrix protocols available at web site http://pathology.wustl.edu/~mgacore/genechip.htm#Preparing. Hybridization of the labeled cRNA to Human Genome U95Av2 chips (Affymetrix) was carried out by using Genechip Instrument System (Affymetrix) at Genechip Core Facility of Washington University. A total of 44 chips were generated and distributed as follows: seven for control pressure (CP) at time 0, four CP at 6 h, seven for hydrostatic pressure (HP) at 6 h, eight for CP at 24 h, eight for HP at 24 h, five for CP at 48 h and five for HP at 48h P. The arrays were washed and stained with streptavidin-phycoerythrin followed by scanning on an Agilent GeneArray Scanner G2500A (Agilent Technologies, Palo Alto, CA), and then scanned by an Affymetrix GeneArray Scanner. Data was analyzed by Affymetrix Microarray Suite (version 5.0), linear regression analysis, and GeneSpring expression Analysis Software (version 6.0, Silicon Genetics). For analysis, the samples were scaled to the same target intensity (1500) to allow comparison of multiple samples, and raw data were normalized to median of each gene across all chips for fold change analysis using GeneSpring. Keywords: time-course
Project description:Transcription profiling of mouse oocytes treated with 20 MPa hydrostatic pressure for 60 minutes at 37 °C comparing control oocytes kept under identical conditions as pressure treated ones, except HHP treatment.
Project description:The GLD-2 class of poly(A) polymerases regulate the timing of translation of stored transcripts by elongating the poly(A) tails of target mRNAs in the cytoplasm. WISPY is a GLD-2 enzyme that acts in the Drosophila female germline and is required for the completion of the egg-to-embryo transition. Though a handful of WISPY target mRNAs have been identified during both oogenesis and early embryogenesis, we aimed to discover the full range of WISPY targets at each stage. To globally identify these targets, we carried out microarray analysis to look for maternal mRNAs whose poly(A) tails fail to elongate in the absence of WISP function. We examined the polyadenylated portion of the maternal transcriptome in both stage 14 (mature) oocytes and in early embryos that had completed egg activation. Our analysis shows that the poly(A) tails of thousands of maternal mRNAs fail to elongate in wisp-deficient oocytes and embryos. Furthermore, we have identified specific classes of genes that are highly regulated in this manner at each stage. Our study shows that cytoplasmic polyadenylation is a major regulatory mechanism during oocyte maturation and egg activation. Four groups of comparisons: WT vs. wisp total RNA from stage 14 oocytes, WT vs. wisp total RNA from fertilized eggs, WT vs. wisp poly(A)+ RNA from stage 14 oocytes, WT vs. wisp poly(A)+ RNA from fertilized eggs. Each comparison consisted of three independent RNA extractions and each experiment was done with dye-swap pairs as two technical replicates.