Project description:Insulin like growth factor binding protein 7, Igfbp7, is a secreted protein that in addition to modulating insulin and insulin-like growth factor signaling, it acts as a tumor suppressor gene in breast and other cancers. To elucidate the role of Igfbp7 in regulating the proliferation and differentiation of mammary epithelial cells, we examined the growth and differentiation of mammary gland through different stages of its development in Igfbp7-null mice. Using transcriptome profiling in addition to functional assays we demonstrate that loss of Igfbp7 leads to diminished luminal cell differentiation and expansion of the luminal progenitors. These studies identify the endocrine factor Igfbp7, as a key regulator of luminal progenitor functions in the mammary gland. Mammary gland mRNA profiles of lactation day 3 wild type (WT) and Igfbp7-/- (KO) mice were generated by deep sequencing using SOLiD5500xl
Project description:Scn1b null mice are a model of a severe developmental and epileptic encephalopathy called Dravet Syndrome (DS). The goal of this study was to identify changes in gene expression between Scn1b wild-type and Scn1b null mice before seizure onset (postnatal day 10)
Project description:Scn1b null mice are a model of a severe developmental and epileptic encephalopathy called Dravet Syndrome (DS). The goal of this study was to identify changes in gene expression between Scn1b wild-type and Scn1b null mice before seizure onset (postnatal day 10) in cortical layer VI, a region known to have differences in excitability in Scn1b null mice. RNA-Seq identified 21 genes, primarily extracellular matrix genes, which were differentially expressed between the two genotypes.
Project description:Transcriptional profiling of 13.5 day mouse embryo forelimbs. Gene expression comparison done between wild type and Chsy-1 gene knockout mice.
Project description:The neocortex and striatum were harvested from conditional Met null male mice (Metfx/fx/nestincre) and wild type littermates at postnatal day 14. Synaptosomes were generated and prepared from each region for quantitative proteomic analyses using isobaric tags for relative and absolute quantitation (iTRAQ).
Project description:Purpose: To assess global changes to the levels of mRNA and protein in butyrophilin-null mice (Btn1a1-/-), which were previously shown to be defective in regulated milk-lipid secretion in the lactating mammary gland. Methods: mRNA profiles of wild-type (WT) and Btn1a1-/-mice were generated by mRNA/cDNA library preparation (Illumina), qPCR quantitation, cluster generation and pair-end deep sequencing (Illumina HiSeq 4000 sequencer) using an Illumina HiSeq SBS kit. Changes in mRNA levels (null/WT ≥ ± 2-fold; adjusted p ≤ 0.05) were assesed by GSEA and IPA analysis and compared with a proteomic analysis of the same sample set.. Results: Samples had 95-167 million pass filter reads (92% above Q30). Mapping of all samples was > 97% (mm10 reference genome). Significant changes in mRNA amounts accounted for 1.4% of 11,834 transcipts (113 up and 53 down regulated). Only 10 mRNA/protein pairs were significantly upregulated and one (Btn1a1) downregulated. Pathways and networks associated with inflammation, apoptosis and the cell cycle were upregulated and lipid synthesis and metabolism were downregulated. Biochemical and histochemical analysis confirmed that lactation is maintained in null animals, in the face of significant apoptosis, by the continual regeneration of secretory epithelial cells. Cell death proceeded through several pathways including activation of caspases 8 and 3 and not by the lysosomal lysis pathway predominant in WT cells Conclusions: Ablation of Btn1a1 has multiple effects on milk-secreting mammary cells, including disruption of lipid secretion, increased cell damage and death. Lactation is maintained by continuous regeneration of the epithelium. Expression of functional Btn1a1 is essential for the maintainance of terminally differentiated mammary cells and optimal milk production throughout lactation.
Project description:We performed mRNA transcriptional profiling of mouse retina in the wild-type and Nrl-null context to determine Nrl-dependent gene expression We sequenced cDNA libraries made from polyA+ selected RNA from retinas of litter-matched WT and Nrl-/- adult mice at postnatal day 21 (P21) (WT vs Nrl-/-, n=5 and n= 6 resp.)
Project description:Insulin like growth factor binding protein 7, Igfbp7, is a secreted protein that in addition to modulating insulin and insulin-like growth factor signaling, it acts as a tumor suppressor gene in breast and other cancers. To elucidate the role of Igfbp7 in regulating the proliferation and differentiation of mammary epithelial cells, we examined the growth and differentiation of mammary gland through different stages of its development in Igfbp7-null mice. Using transcriptome profiling in addition to functional assays we demonstrate that loss of Igfbp7 leads to diminished luminal cell differentiation and expansion of the luminal progenitors. These studies identify the endocrine factor Igfbp7, as a key regulator of luminal progenitor functions in the mammary gland.
Project description:Previoulsly expression profiling of the whole mammary gland across different stages of pregnancy and lactation has been performed on different strains of mice. Since mammary gland has both epithelial and stromal compartments, to specifically identify the genes involved in the transition from pregnancy to lactation a process termed as secretory activation, expression profiling of isolated mammary epithelial cells (MECs) from four CD1 mice each at Pregnancy day 14 (P14) and Lactation day 2 (L2) was performed in the current study. Statistical analysis of the mRNA changes between P14 and L2 identified 5,499 unique genes as being differentially expressed (5% FDR), of which, 2,902 genes and 2,604 genes were higher in P14 or L2 stages, respectively.
Project description:Multidrug resistance related protein 4 (MRP4) belongs to the C subfamily of ABC transporters and maintains cellular environment by functioning as an efflux pump. We aimed to obtain insights into the coeffects of MRP4 deficiency and ageing on the retina, and gene expression profiling using DNA microarray were performed. To find out genes significantly up- or downregulated in aged Mrp4-null mice, the comparison of microarray data using retinal samples was carried out between aged Mrp4-null mice and aged wild-type mice.
