Simultaneous microRNA and mRNA expression profiling on mammary epithelial cells isolated from mice at pregnancy day 14 and lactation day 2
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ABSTRACT:
ORGANISM(S): synthetic construct Mus musculus
PROVIDER: GSE87584 | GEO | 2016/10/05
SECONDARY ACCESSION(S): PRJNA345241
REPOSITORIES: GEO
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Project description:Simultaneous microRNA and mRNA expression profiling on mammary epithelial cells isolated from mice at pregnancy day 14 and lactation day 2
| PRJNA345241 | ENA
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.
2016-10-05 | GSE73837 | GEO
Project description:Previoulsly miRNA expression profiling of the whole mammary gland across different stages of pregnancy and lactation has been performed in mice. Since mammary gland has both epithelial and stromal compartments, to specifically identify the miRNAs 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 miRNA changes between P14 and L2 identified 32 miRNAs to be differentially expressed with a fold change greater than or equal to 2, of which, the majority of them declinied at the onset of lactation.
2016-10-05 | GSE79949 | GEO
Project description:BackgroundDuring pregnancy, as the mammary gland prepares for synthesis and delivery of milk to newborns, a luminal mammary epithelial cell (MEC) subpopulation proliferates rapidly in response to systemic hormonal cues that activate STAT5A. While the receptor tyrosine kinase ErbB4 is required for STAT5A activation in MECs during pregnancy, it is unclear how ErbB3, a heterodimeric partner of ErbB4 and activator of phosphatidyl inositol-3 kinase (PI3K) signaling, contributes to lactogenic expansion of the mammary gland.MethodsWe assessed mRNA expression levels by expression microarray of mouse mammary glands harvested throughout pregnancy and lactation. To study the role of ErbB3 in mammary gland lactogenesis, we used transgenic mice expressing WAP-driven Cre recombinase to generate a mouse model in which conditional ErbB3 ablation occurred specifically in alveolar mammary epithelial cells (aMECs).ResultsProfiling of RNA from mouse MECs isolated throughout pregnancy revealed robust Erbb3 induction during mid-to-late pregnancy, a time point when aMECs proliferate rapidly and undergo differentiation to support milk production. Litters nursed by ErbB3 KO dams weighed significantly less when compared to litters nursed by ErbB3 WT dams. Further analysis revealed substantially reduced epithelial content, decreased aMEC proliferation, and increased aMEC cell death during late pregnancy. Consistent with the potent ability of ErbB3 to activate cell survival through the PI3K/Akt pathway, we found impaired Akt phosphorylation in ErbB3 KO samples, as well as impaired expression of STAT5A, a master regulator of lactogenesis. Constitutively active Akt rescued cell survival in ErbB3-depleted aMECs, but failed to restore STAT5A expression or activity. Interestingly, defects in growth and survival of ErbB3 KO aMECs as well as Akt phosphorylation, STAT5A activity, and expression of milk-encoding genes observed in ErbB3 KO MECs progressively improved between late pregnancy and lactation day 5. We found a compensatory upregulation of ErbB4 activity in ErbB3 KO mammary glands. Enforced ErbB4 expression alleviated the consequences of ErbB3 ablation in aMECs, while combined ablation of both ErbB3 and ErbB4 exaggerated the phenotype.ConclusionsThese studies demonstrate that ErbB3, like ErbB4, enhances lactogenic expansion and differentiation of the mammary gland during pregnancy, through activation of Akt and STAT5A, two targets crucial for lactation.
| S-EPMC5591538 | biostudies-literature
Project description:The level of gamma-glutamyltranspeptidase (GGT) activity and of its mRNA were determined in the mouse mammary gland during pregnancy, lactation and weaning. The GGT activity, which is very low in the virgin-mouse mammary gland (5 munits/mg of protein), increases progressively during pregnancy (3-fold), reaches its maximum at the onset of lactation (8-fold) and returns rapidly to basal level at weaning. Although no GGT-specific mRNA is detected in the virgin-mouse mammary gland, a single faint band of 2.2 kb in size is found during pregnancy. During lactation, an additional mRNA of 2.4 kb in size appears, and the level of both mRNAs is higher. This high level of mRNA persists during weaning as well. Southern-blot analysis of mouse mammary-gland DNA provides convincing evidence that there is only one gene which codes for the two mRNAs. The present study provides the first evidence for a physiological regulation of the two GGT mRNAs in the same tissue.
| S-EPMC1131342 | biostudies-other
Project description:The calmodulin content of heat-treated extracts of rat mammary tissue and isolated cells was measured by using stimulation of cyclic nucleotide phosphodiesterase (PDE) activity and radioimmunoassay (r.i.a.) procedures. The calmodulin content of mammary tissue increased 2.5-fold near the time of parturition, remained at the elevated level during lactation, then, after the onset of involution, decreased to values similar to those measured from mammary tissue of pregnant rats. When tissue from 15 animals in different stages of pregnancy, lactation and involution were compared, the r.i.a. gave 2.6-fold higher results than the PDE assay. To investigate further the increase in calmodulin content of mammary tissue, secretory and myoepithelial cells were enzymically dissociated from rat mammary tissue during different stages of pregnancy, lactation and involution. Protein, DNA, lactose, glucose-6-phosphate dehydrogenase and alkaline phosphatase were assayed to characterize the cell fractions. By using r.i.a., the calmodulin content per mg of protein in isolated secretory-cell fractions was high near parturition, then decreased and remained relatively constant during lactation. The amount of calmodulin expressed per mg of DNA in secretory cells did not show a marked change near parturition, suggesting a constant amount of calmodulin per cell. The calmodulin content of myoepithelial cells dissociated from mammary tissue and measured by using r.i.a. was 6-fold lower than in secretory cells and remained relatively constant during the course of lactation. The changing levels of calmodulin in rat mammary tissue during development are suggested to be related to proliferation and destruction of secretory epithelial cells, events that occur near parturition and involution respectively.
| S-EPMC1153563 | biostudies-other
Project description:Our study revealed that CUZD1 (CUB and zona pellucida-like domain containing protein-1) is a novel target of estrogen regulation in the mouse mammary epithelium. Mice lacking Cuzd1 exhibit delayed ductal outgrowth during puberty and a striking impairment in ductal branching and alveolar development during pregnancy. Ablation of Cuzd1 led to a marked reduction in steroid-induced proliferation of mammary ductal and alveolar epithelium. To identify the downstream targets of Cuzd1 in mammary gland development, we performed gene expression profling of mammary epithlial cells isolated from Cuzd1-null and its heterozygous litteremates on day 18 of pregnancy. The microarray results revealed downregulation of mRNAs corresponding to several members of the epidermal growth factor family in mammary epithelial cells of Cuzd1-null mice. Consequently, the activation of the ERBB receptors, ERBB1 and ERBB4, and their downstream target STAT5, was disrupted in Cuzd1-null mammary tissue. Collectively, these findings support a unique role for CUZD1 as a critical mediator of the steroid-induced proliferation and differentiation of ductal epithelium during pregnancy and lactation.
2014-01-01 | GSE30939 | GEO
Project description:Activity of glucocorticoid receptor in mouse mammary cytosol changes during lactogenesis. The highest receptor activity is observed in the second half of pregnancy. The receptor from mammary glands from lactating and pregnant mice exhibits the same sedimentation pattern, as well as the same specificity and affinity for binding of steroid hormones.
| S-EPMC1163992 | biostudies-other
Project description:Our study revealed that CUZD1 (CUB and zona pellucida-like domain containing protein-1) is a novel target of estrogen regulation in the mouse mammary epithelium. Mice lacking Cuzd1 exhibit delayed ductal outgrowth during puberty and a striking impairment in ductal branching and alveolar development during pregnancy. Ablation of Cuzd1 led to a marked reduction in steroid-induced proliferation of mammary ductal and alveolar epithelium. To identify the downstream targets of Cuzd1 in mammary gland development, we performed gene expression profling of mammary epithlial cells isolated from Cuzd1-null and its heterozygous litteremates on day 18 of pregnancy. The microarray results revealed downregulation of mRNAs corresponding to several members of the epidermal growth factor family in mammary epithelial cells of Cuzd1-null mice. Consequently, the activation of the ERBB receptors, ERBB1 and ERBB4, and their downstream target STAT5, was disrupted in Cuzd1-null mammary tissue. Collectively, these findings support a unique role for CUZD1 as a critical mediator of the steroid-induced proliferation and differentiation of ductal epithelium during pregnancy and lactation. To investigate the functional role of Cuzd1, we created Cuzd1-null mice by homologous recombination using mouse embryonic stem cells. As severe defect was observed in alveolargenesis and lactation of Cuzd1 null mammary gland, we purified mammary epithlial cells from day18 pregnant mice (n=5 for each genotype), purified total RNA from these cells, pooled these samples and then hybridized to high density affymetrix microarrays.
2014-01-01 | E-GEOD-30939 | biostudies-arrayexpress
Project description:Polyamines and RNA accumulate in the rat mammary gland during pregnancy, but the major increases occur after parturition. Therefore the major increases occur after the gland has obtained its maximal complement of epithelial cells. During lactation, the spermidine concentration rises above 5mm and RNA content in the lactating mammary gland reaches a value 16 times that of the unstimulated mammary gland. The ratio of spermidine/spermine, an increase of which initially signals an elevation in biosynthetic activity, is near 1 in the normal mammary gland and is greater than 10 in the lactating mammary gland. Putrescine concentration is very low during the entire course of mammary-gland development, with the exception of early pregnancy. The low putrescine concentration probably reflects the very rapid conversion of putrescine into spermidine. Both ornithine decarboxylase, the enzyme that synthesizes putrescine, and putrescine-stimulated S-adenosyl-l-methionine decarboxylase, the enzyme that synthesizes spermidine, increase in activity during middle and late pregnancy; during lactation, both enzyme activities are elevated until the 21st day of lactation, and then decline. These declines are concomitant with involution. Also, it was found that the amount of ribonuclease activity in the mammary gland was very high during lactation, almost double that in the gland during pregnancy.
| S-EPMC1174302 | biostudies-other