Project description:We constructed a comparative proteome profile of female mouse fetal gonads at specific time points (11.5, 12.5, and 13.5 days post coitum), spanning a critical window for initiation of meiosis in female germ cells. We identified 3666 proteins, of which 473 were differentially expressed.
Project description:Lmx1b regulates dorsalization of limb fates, but the mechanism of this regulation has not been characterized. To identify candidate genes regulated by Lmx1b we compared the limbs from Lmx1b KO mice to wild type mice during limb dorsalization (e11.5-13.5). Differentially expressed genes that we common to all three stages examined were considered to be likely candidates for Lmx1b regulation and further evaluated. At 11.5 and 12.5 dpc, embryos were harvested and the limb buds with the limb girdles were isolated. Embryos at 13.5dpc were also harvested and their distal limb buds (zeugopods and autopods) were isolated. Embryos were genotyped to confirm Lmx1b homozygosity (-/- or +/+). RNA from embryonic forelimbs and hindlimbs of wild type (WT) and Lmx1b KO mice was harvested using the Rneasy Kit (Qiagen). RNA was pooled to decrease genetic variability, i.e., six limbs at 11.5 dpc, three limbs at 12.5 dpc and six limbs at 13.5 dpc. Duplicate samples were generated using different embryos for each stage and then hybridized to the Affymetrix GeneChip® Mouse Genome 430 2.0 Array (UCI, Irvine, CA).
Project description:The aim of this study was to analyze the gene expression profile for three main cell lines (supporting, interstitial/stromal, and germ cells) isolated from developing gonads at the critical period of sexual differentiation (between 11.5, and 13.5 dpc). Three cell lines (supporting, interstitial/stromal, and germ cells) were isolated from murine fetal XX and XY gonads at three time points (11.5, 12.5, and 13.5 dpc). Transgenic mouse strains with the expression of cell type specific fluorescent markers were used to isolate the cell lines. Cells were sorted using FACS method and then the RNA was extracted.
Project description:Testicular cord formation in male gonadogenesis involves assembly of several cell types, the precise molecular mechanism is still not well known.With the high-throughput quantitative proteomics technology, a comparative proteomic profile of mouse embryonic male gonads were analyzed at three time points (11.5, 12.5 and 13.5 days post coitum), corresponding to critical stages of testicular cord formation in gonadal development.
Project description:The aim of this study was to analyze the gene expression profile for three main cell lines (supporting, interstitial/stromal, and germ cells) isolated from developing gonads at the critical period of sexual differentiation (between 11.5, and 13.5 dpc).
Project description:Mus musculus embryonic submandibular salivary glands from embryonic day 12.5 embryos were laser microdissected to isolate epithelial cells from regions adjacent to forming clefts or from the peripheral end bud region. The isolated region-specific samples were then subjected to T7-SAGE transcriptome analysis.
Project description:Understanding congenital liver disease requires elucidation of the signaling pathways and transcriptional events in the developing liver. Comprehensive assessment of gene expression between 10.5 and 16.5 dpc in the developing mouse liver and comparison with adult liver and non-hepatic embryonic tissue was validated with real-time PCR and in situ hybridization. The broad nature of the analysis provides insights into patterns of genetic control of hepatogenesis. Pathways implicated in human disease are highly regulated at the transcriptional level. Rather than activating or inhibiting a pathway or biological process by altering the expression of a single signaling molecule, transcriptional changes in large numbers of genes in a pathway or process are regulated in a coordinated manner. For example, both TGF-beta and Notch signaling is inhibited during hepatogenesis not just by decreasing transcription of multiple pathway members, but also with a complementary increase in the transcription of a pathway inhibitor. Similarly, genes related to specific biological processes exhibit strong temporal synchronization in which multiple members of the pathway have similar transcriptional regulation over time. Global coordination of signaling or functional families at the transcriptional level may be a mechanism to produce robustness of the desired outcomes. In addition, this comprehensive analysis provides a database for the further study of transcriptional events during liver development by identifying liver-specific, highly regulated genes. Experiment Overall Design: In order to provide transcriptional profile of the developing liver compared both to normal adult liver and non-hepatic embryonic tissueswe performed high-density microarray analysis using Affymetrix MG 430 2.0 chips for embryonic liver samples at 10.5, 11.5, 12.5, 13.5, 14.5, and 16.5 days post conception (dpc), embryo-minus liver tissues at 10.5, 11.5, 12.5, and 14.5 dpc, and normal 10-week-old adult mouse liver. Each sample consisted of at least five embryos.
Project description:Expression profiling of micro RNAs in mouse embryonic gonads before (11.5 days post coitum) and after (13.5 days post coitum) the critical period of sex determination
Project description:We compared gene expression differences in the polytypic species complex Mus musculus (Mus musculus musculus, Mus musculus domesticus, Mus musculus castaneus and Mus musculus ssp) with that of Mus spretus via oligonucleotide microarrays representing more than 20,000 genes. Analysis of the results by two way ANOVA statistics suggests that the most genes with significant differences in expression levels among the subspecies are found in liver and kidney and the least in testis. This picture is different when one compares with Mus spretus, where the largest number of differences is found in testis. Keywords: multi-species comparison
Project description:Whole-genome gene expression analysis was performed using the AGMs from two littermate pairs of tph2+/+, tph2+/- or tph2-/- at E11.5. The total RNA was extracted using TRNzol (Tiangen, Beijing) and cDNA samples were sequenced using Illumina HiSeq3000 (RiboBio Co. Ltd, Guangzhou). The reference Mus musculus genome and gene information were downloaded from the National Center for Biotechnology Information Database (NCBI: http://www.ncbi.nlm.nih.gov/).