Project description:Following intrauterine low protein diet, the lumbar mammary gland of control and low protein rats were removed for further analysis via genarray on day 21 and day 28 postpartum.
Project description:RNA from MMTV-Cre;Sox9flox/flox mouse mammary glands were compared to RNA from MMTV-Cre;Sox9+/flox glands. Results indicate that Sox9 regulates several genes that impact ductal morphogenesis in the mammary gland.
Project description:RNA from MMTV-Cre;Sox9flox/flox mouse mammary glands were compared to RNA from MMTV-Cre;Sox9+/flox glands. Results indicate that Sox9 regulates several genes that impact ductal morphogenesis in the mammary gland. The portion of the fourth mammary gland that is proximal to the intra-mammary gland lymph nodes was dissected from four 5-week-old MMTV-Cre;Sox9flox/flox females and four MMTV-Cre;Sox9+/flox females of the same age. Total RNA from each gland was extracted and hybridized to separate Affymetrix Gene 1.0 ST chips.
Project description:Mammary gland branching morphogenesis is thought to depend on the mobilization of the membrane-anchored matrix metalloproteinases, MT1-MMP and MT2-MMP, that drive epithelial cell invasion by remodeling the extracellular matrix and triggering associated signaling cascades. However, the roles that these proteinases play during mammary gland development in vivo remains undefined. A mammary gland branching program that occurs during the first 10 days of early postnatal development was used to characterize the impact of global Mt1-mmp or Mt2-mmp targeting on mammary gland morphogenesis. Transcriptome profiling of ductal networks and associated stroma was used to investigate the functional roles of MT2-MMP in the early postnatal mammary gland in an unbiased fashion.
Project description:Mammary gland branching morphogenesis is thought to depend on the mobilization of the membrane-anchored matrix metalloproteinases, MT1-MMP and MT2-MMP, that drive epithelial cell invasion by remodeling the extracellular matrix and triggering associated signaling cascades. However, the roles that these proteinases play during mammary gland development in vivo remains undefined. A mammary gland branching program that occurs during the first 10 days of early postnatal development was used to characterize the impact of global Mt1-mmp or Mt2-mmp targeting on mammary gland morphogenesis. Transcriptome profiling of ductal networks and associated stroma was used to investigate the functional roles of MT1-MMP in the early postnatal mammary gland in an unbiased fashion.
Project description:Mammary gland branching morphogenesis is thought to depend on the mobilization of proteolytic machinery from the matrix metalloproteinase (MMP) family, namely MT1-MMP/MMP14, to drive coordinated epithelial cell invasion through the interstitial extracellular matrix, but the dominant effector has remained undefined. Unexpectedly, we find MMP14 controls postnatal mammary gland branching from the periductal stroma. Transcriptome profiling of stromal cell-targeted mammary glands was used to characterize the impact of stromal Mmp14-targeting on the growth factor and signaling cascades implicated in mammary gland morphogenesis. Transcriptome profiling of ductal networks and associated stroma was used to investigate the functional roles of MMP14 in the postnatal mammary gland stroma in an unbiased fashion.
Project description:Following intrauterine low protein diet, the lumbar mammary gland of control and low protein rats were removed for further analysis via genarray on day 21 and day 28 postpartum. Virgin female Wistar rats came from Charles River. Throughout pregnancy the NP rats received 25g/d of Altromin C1000 containing 17.0% protein, while LP animals were fed 25g/d of Altromin C1003 containing 8.8% protein. At birth, litter size was reduced to 6 females in both groups, which were left to suckle for 21 days. During lactation (d1-d21) and after weaning (d21-d28) NP and LP animals were fed a standard chow and tap water ad libitum. At d21 and d28 3 dams were sacrificed under isofluran narcosis. The lumbar mammary gland (4th and 5th pair) were removed and cleared of visible lymph nodes before snap-freezing in liquid nitrogen.
Project description:The identification of genes driving organ development is central to understanding which signaling pathways drive the pathogenesis of various diseases including cancer. This dataset depicts the proteomic changes observed in C57BL/6J mice expressing wild-type or 3SA-phospho mutant versions of the Bcl-2-associated death promoter, BAD. This data shows that BAD regulates postnatal mammary gland morphogenesis in puberty. Three conserved serine residues on BAD are co-ordinately phosphorylated to regulate its activity. Non-phosphorylated BAD mutant delayed pubertal ductal elongation. This defect was specific to the epithelial compartment as transplant and ex vivo organoid assays of mutant epithelium recapitulated decreased tubule migration. Proteomic signature between BAD+/+ and phosphomutant BAD-3SA mammary glands identified differences in actin-binding and focal adhesion components. Mechanistically, non-phosphorylated BAD impedes protein translation, specifically in protrusions, through aberrant hypophosphorylated 4E-BP1. These findings reveal a critical enhancement of localized translation for efficient pubertal-mammary-gland morphogenesis and identifies BAD as a novel regulator of this process.
Project description:Mammary gland ductal morphogenesis depends on the differentiation of mammary stem cells (MaSCs) into basal and luminal lineages. The AP-2γ transcription factor, encoded by Tfap2c, has a central role in mammary gland development but its effect in mammary lineages and specifically MaSCs is largely unknown. Herein, we utilized an inducible, conditional knockout of Tfap2c to elucidate the role of AP-2γ in maintenance and differentiation of MaSCs. Loss of AP-2γ in the basal epithelium profoundly altered the transcriptomes and decreased the number of cells within several clusters of mammary epithelial cells, including adult MaSCs and luminal progenitors. AP-2γ regulated the expression of genes known to be required for mammary development including Cebpb, Nfkbia, and Rspo1. As a result, AP-2γ-deficient mice exhibited repressed mammary gland ductal outgrowth and inhibition of regenerative capacity. The findings demonstrate that AP-2γ can regulate development of mammary gland structures potentially regulating maintenance and differentiation of multipotent MaSCs.
Project description:Macrophages are diverse immune cells that reside in all tissues to regulate development, function and homeostasis. In the mammary gland, a highly dynamic organ that undergoes dramatic remodeling throughout life, macrophages have been implicated in development and breast tumorigenesis. Ductal macrophages, as opposed to their stromal counterparts, were found to promote ductal morphogenesis during puberty and efficiently phagocytose alveolar cells post-lactation. Within mammary tumors, only ductal macrophages expanded to form a network throughout the tumor, where they also contact intra-tumoral T cells. Furthermore, ductal macrophages accelerated the growth of tumor organoids. Thus, pro-remodeling mammary ductal macrophages represent the normal tissue counterpart of tumor macrophages in an organ that is highly susceptible to oncogenesis. To gain insight into ductal macrophage function, we sorted all antigen presenting cell populations from mouse mammary glands and undertook RNA-seq expression profiling. This showed that ductal macrophages have a distinct expression profile from those of stromal macrophages or dendritic cells from the mammary gland. Results from this data series are presented in Figure 2 of Dawson et al (2020).