Vitamin D3 receptor ablation alters mammary gland morphogenesis

Development ◽  
2002 ◽  
Vol 129 (13) ◽  
pp. 3067-3076
Author(s):  
Glendon Zinser ◽  
Kathryn Packman ◽  
JoEllen Welsh
Keyword(s):  
Mammary Gland ◽  
Vitamin D3 ◽  
Stromal Cells ◽  
Knockout Mice ◽  
Wild Type ◽  
Dihydroxyvitamin D3 ◽  
Mammary Gland Morphogenesis ◽  
Gland Morphogenesis ◽  
Gland Development

Postnatal mammary gland morphogenesis is achieved through coordination of signaling networks in both the epithelial and stromal cells of the developing gland. While the major proliferative hormones driving pubertal mammary gland development are estrogen and progesterone, studies in transgenic and knockout mice have successfully identified other steroid and peptide hormones that impact on mammary gland development. The vitamin D3 receptor (VDR), whose ligand 1,25-dihydroxyvitamin D3 is the biologically active form of vitamin D3, has been implicated in control of differentiation, cell cycle and apoptosis of mammary cells in culture, but little is known about the physiological relevance of the vitamin D3 endocrine system in the developing gland. In these studies, we report the expression of the VDR in epithelial cells of the terminal end bud and subtending ducts, in stromal cells and in a subset of lymphocytes within the lymph node. In the terminal end bud, a distinct gradient of VDR expression is observed, with weak VDR staining in proliferative populations and strong VDR staining in differentiated populations. The role of the VDR in ductal morphogenesis was examined in Vdr knockout mice fed high dietary Ca2+ which normalizes fertility, serum estrogen and neonatal growth. Our results indicate that mammary glands from virgin Vdr knockout mice are heavier and exhibit enhanced growth, as evidenced by higher numbers of terminal end buds, greater ductal outgrowth and enhanced secondary branch points, compared with glands from age- and weight-matched wild-type mice. In addition, glands from Vdr knockout mice exhibit enhanced growth in response to exogenous estrogen and progesterone, both in vivo and in organ culture, compared with glands from wild-type mice. Our data provide the first in vivo evidence that 1,25-dihydroxyvitamin D3 and the VDR impact on ductal elongation and branching morphogenesis during pubertal development of the mammary gland. Collectively, these results suggest that the vitamin D3 signaling pathway participates in negative growth regulation of the mammary gland.

scholarly journals The Ras-like protein R-Ras2/TC21 is important for proper mammary gland development

2012 ◽  
Vol 23 (12) ◽  
pp. 2373-2387 ◽  
Author(s):  
Romain M. Larive ◽  
Antonio Abad ◽  
Clara M. Cardaba ◽  
Teresa Hernández ◽  
Marta Cañamero ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Knockout Mice ◽  
Physiological Role ◽  
Wild Type ◽  
Erk Activation ◽  
Terminal End Buds ◽  
Phosphorylated Akt ◽  
Morphogenic Process ◽  
Gland Development

R-Ras2/TC21 is a GTPase with high sequence and signaling similarity with Ras subfamily members. Although it has been extensively studied using overexpression studies in cell lines, its physiological role remains poorly characterized. Here we used RRas2-knockout mice expressing β-galactosidase under the regulation of the endogenous RRas2 promoter to investigate the function of this GTPase in vivo. Despite its expression in tissues critical for organismal viability, RRas2−/− mice show no major alterations in viability, growth rates, cardiovascular parameters, or fertility. By contrast, they display a marked and specific defect in the development of the mammary gland during puberty. In the absence of R-Ras2/TC21, this gland forms reduced numbers of terminal end buds (TEBs) and ductal branches, leading to a temporal delay in the extension and arborization of the gland tree in mammary fat pads. This phenotype is linked to cell-autonomous proliferative defects of epithelial cells present in TEBs. These cells also show reduced Erk activation but wild type–like levels of phosphorylated Akt. Using compound RRas2-, HRas-, and NRas-knockout mice, we demonstrate that these GTPases act in a nonsynergistic and nonadditive manner during this morphogenic process.

scholarly journals Insights into the role of connexins in mammary gland morphogenesis and function

Reproduction ◽  
2015 ◽  
Vol 149 (6) ◽  
pp. R279-R290 ◽  
Author(s):  
Michael K G Stewart ◽  
Jamie Simek ◽  
Dale W Laird
Keyword(s):  
Mammary Gland ◽  
Gap Junctions ◽  
Normal Tissue ◽  
Mammary Gland Development ◽  
Mammary Gland Morphogenesis ◽  
Gland Morphogenesis ◽  
Cell Death Mechanisms ◽  
And Function ◽  
Gland Development

Gap junctions formed of connexin subunits link adjacent cells by direct intercellular communication that is essential for normal tissue homeostasis in the mammary gland. The mammary gland undergoes immense remodeling and requires exquisite regulation to control the proliferative, differentiating, and cell death mechanisms regulating gland development and function. The generation of novel genetically modified mice with reduced or ablated connexin function within the mammary gland has advanced our understanding of the role of gap junctions during the complex and dynamic process of mammary gland development. These studies have revealed an important stage-specific role for Cx26 (GJA1) and Cx43 (GJB2), while Cx30 (GJB6) and Cx32 (Gjb1) can be eliminated without compromising the gland. Yet, there remain gaps in our understanding of the role of mammary gland gap junctions.

scholarly journals Essential function of Wnt-4 in mammary gland development downstream of progesterone signaling

2000 ◽  
Vol 14 (6) ◽  
pp. 650-654 ◽  
Author(s):  
Cathrin Brisken ◽  
Anna Heineman ◽  
Tony Chavarria ◽  
Brian Elenbaas ◽  
Jian Tan ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Mammary Epithelial Cells ◽  
Ectopic Expression ◽  
Mammary Epithelium ◽  
Reproductive Hormones ◽  
Mammary Epithelial ◽  
Mammary Gland Morphogenesis ◽  
Mammary Epithelia ◽  
Gland Morphogenesis ◽  
Gland Development

Female reproductive hormones control mammary gland morphogenesis. In the absence of the progesterone receptor (PR) from the mammary epithelium, ductal side-branching fails to occur. We can overcome this defect by ectopic expression of the protooncogene Wnt-1. Transplantation of mammary epithelia fromWnt-4−/− mice shows that Wnt-4 has an essential role in side-branching early in pregnancy. PR andWnt-4 mRNAs colocalize to the luminal compartment of the ductal epithelium. Progesterone induces Wnt-4 in mammary epithelial cells and is required for increased Wnt-4 expression during pregnancy. Thus, Wnt signaling is essential in mediating progesterone function during mammary gland morphogenesis.

scholarly journals Cyclin-Dependent Kinase Inhibitor P27Kip1 Is Required for Mouse Mammary Gland Morphogenesis and Function

2001 ◽  
Vol 153 (5) ◽  
pp. 917-932 ◽  
Author(s):  
Rebecca S. Muraoka ◽  
Anne E.G. Lenferink ◽  
Jean Simpson ◽  
Dana M. Brantley ◽  
L. Renee Roebuck ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Cyclin D1 ◽  
Mammary Epithelial Cells ◽  
Mammary Glands ◽  
Mammary Epithelial ◽  
Cyclin Dependent Kinase ◽  
Wild Type ◽  
Mammary Gland Morphogenesis ◽  
Ductal Branching ◽  
Gland Morphogenesis

We have studied the role of the cyclin-dependent kinase (Cdk) inhibitor p27Kip1 in postnatal mammary gland morphogenesis. Based on its ability to negatively regulate cyclin/Cdk function, loss of p27 may result in unrestrained cellular proliferation. However, recent evidence about the stabilizing effect of p27 on cyclin D1–Cdk4 complexes suggests that p27 deficiency might recapitulate the hypoplastic mammary phenotype of cyclin D1–deficient animals. These hypotheses were investigated in postnatal p27-deficient (p27−/−), hemizygous (p27+/−), or wild-type (p27+/+) mammary glands. Mammary glands from p27+/− mice displayed increased ductal branching and proliferation with delayed postlactational involution. In contrast, p27−/− mammary glands or wild-type mammary fat pads reconstituted with p27−/− epithelium produced the opposite phenotype: hypoplasia, low proliferation, decreased ductal branching, impaired lobuloalveolar differentiation, and inability to lactate. The association of cyclin D1 with Cdk4, the kinase activity of Cdk4 against pRb in vitro, the nuclear localization of cyclin D1, and the stability of cyclin D1 were all severely impaired in p27−/− mammary epithelial cells compared with p27+/+ and p27+/− mammary epithelial cells. Therefore, p27 is required for mammary gland development in a dose-dependent fashion and positively regulates cyclin D–Cdk4 function in the mammary gland.

scholarly journals BAD regulates mammary gland morphogenesis by 4E-BP1-mediated control of localized translation in mouse and human models

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
John Maringa Githaka ◽  
Namita Tripathi ◽  
Raven Kirschenman ◽  
Namrata Patel ◽  
Vrajesh Pandya ◽  
...  
Keyword(s):  
Cell Migration ◽  
Mammary Gland ◽  
Mammary Gland Development ◽  
Focal Adhesions ◽  
Mrna Translation ◽  
Protein Functions ◽  
Mammary Gland Morphogenesis ◽  
Gland Morphogenesis ◽  
Gland Development

AbstractElucidation of non-canonical protein functions can identify novel tissue homeostasis pathways. Herein, we describe a role for the Bcl-2 family member BAD in postnatal mammary gland morphogenesis. In Bad3SA knock-in mice, where BAD cannot undergo phosphorylation at 3 key serine residues, pubertal gland development is delayed due to aberrant tubulogenesis of the ductal epithelium. Proteomic and RPPA analyses identify that BAD regulates focal adhesions and the mRNA translation repressor, 4E-BP1. These results suggest that BAD modulates localized translation that drives focal adhesion maturation and cell motility. Consistent with this, cells within Bad3SA organoids contain unstable protrusions with decreased compartmentalized mRNA translation and focal adhesions, and exhibit reduced cell migration and tubulogenesis. Critically, protrusion stability is rescued by 4E-BP1 depletion. Together our results confirm an unexpected role of BAD in controlling localized translation and cell migration during mammary gland development.

Mammary gland morphogenesis is inhibited in transgenic mice that overexpress cell surface beta1,4-galactosyltransferase

Development ◽  
1996 ◽  
Vol 122 (9) ◽  
pp. 2859-2872 ◽  
Author(s):  
H.J. Hathaway ◽  
B.D. Shur
Keyword(s):  
Mammary Gland ◽  
Cell Surface ◽  
Transgenic Animals ◽  
Mammary Epithelial ◽  
Wild Type ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Mammary Gland Morphogenesis ◽  
Gland Morphogenesis ◽  
Cell Matrix Interactions

Mammary gland morphogenesis is facilitated by a precise sequence of cell-cell and cell-matrix interactions, which are mediated in part through a variety of cell surface receptors and their ligands (Boudreau, N., Myers, C. and Bissell, M. J. (1995). Trends in Cell Biology 5, 1–4). Cell surface beta1,4-galactosyltransferase (GalTase) is one receptor that participates in a variety of cell-cell and cell-matrix interactions during fertilization and development, including mammary epithelial cell-matrix interactions (Barcellos-Hoff, M. H. (1992). Exp. Cell Res. 201, 225–234). To analyze GalTase function during mammary gland morphogenesis in vivo, we created transgenic animals that overexpress the long isoform of GalTase under the control of a heterologous promoter. As expected, mammary epithelial cells from transgenic animals had 2.3 times more GalTase activity on their cell surface than did wild-type cells. Homozygous transgenic females from multiple independent lines failed to lactate, whereas transgenic mice overexpressing the Golgi-localized short isoform of GalTase lactated normally. Glands from transgenic females overexpressing surface GalTase were characterized by abnormal and reduced ductal development with a concomitant reduction in alveolar expansion during pregnancy. The phenotype was not due to a defect in proliferation, since the mitotic index for transgenic and wild-type glands was similar. Morphological changes were accompanied by a dramatic reduction in the expression of milk-specific proteins. Immunohistochemical markers for epithelia and myoepithelia demonstrated that both cell types were present. To better understand how overexpression of surface GalTase impairs ductal morphogenesis, primary mammary epithelial cultures were established on basement membranes. Cultures derived from transgenic mammary glands were unable to form anastomosing networks of epithelial cells and failed to express milk-specific proteins, unlike wild-type mammary cultures that formed epithelial tubules and expressed milk proteins. Our results suggest that cell surface GalTase is an important mediator of mammary cell interaction with the extracellular matrix. Furthermore, perturbing surface GalTase levels inhibits the expression of mammary-specific gene products, implicating GalTase as a component of a receptor-mediated signal transduction pathway required for normal mammary gland differentiation.

scholarly journals Netrin-1/Neogenin Interaction Stabilizes Multipotent Progenitor Cap Cells during Mammary Gland Morphogenesis

2003 ◽  
Vol 4 (3) ◽  
pp. 371-382 ◽  
Author(s):  
Karpagam Srinivasan ◽  
Phyllis Strickland ◽  
Ana Valdes ◽  
Grace C Shin ◽  
Lindsay Hinck
Keyword(s):  
Mammary Gland ◽  
Mammary Gland Morphogenesis ◽  
Gland Morphogenesis

scholarly journals Preoperative administration of the 5-HT4 receptor agonist prucalopride reduces intestinal inflammation and shortens postoperative ileus via cholinergic enteric neurons

Gut ◽  
2018 ◽  
Vol 68 (8) ◽  
pp. 1406-1416 ◽  
Author(s):  
Nathalie Stakenborg ◽  
Evelien Labeeuw ◽  
Pedro J Gomez-Pinilla ◽  
Sebastiaan De Schepper ◽  
Raymond Aerts ◽  
...  
Keyword(s):  
Knockout Mice ◽  
Postoperative Ileus ◽  
Intestinal Inflammation ◽  
Electrical Field Stimulation ◽  
Enteric Neurons ◽  
Postoperative Treatment ◽  
Wild Type ◽  
Clinical Recovery ◽  
Preoperative Administration

ObjectivesVagus nerve stimulation (VNS), most likely via enteric neurons, prevents postoperative ileus (POI) by reducing activation of alpha7 nicotinic receptor (α7nAChR) positive muscularis macrophages (mMφ) and dampening surgery-induced intestinal inflammation. Here, we evaluated if 5-HT4 receptor (5-HT4R) agonist prucalopride can mimic this effect in mice and human.DesignUsing Ca2+ imaging, the effect of electrical field stimulation (EFS) and prucalopride was evaluated in situ on mMφ activation evoked by ATP in jejunal muscularis tissue. Next, preoperative and postoperative administration of prucalopride (1–5 mg/kg) was compared with that of preoperative VNS in a model of POI in wild-type and α7nAChR knockout mice. Finally, in a pilot study, patients undergoing a Whipple procedure were preoperatively treated with prucalopride (n=10), abdominal VNS (n=10) or sham/placebo (n=10) to evaluate the effect on intestinal inflammation and clinical recovery of POI.ResultsEFS reduced the ATP-induced Ca2+ response of mMφ, an effect that was dampened by neurotoxins tetrodotoxin and ω-conotoxin and mimicked by prucalopride. In vivo, prucalopride administered before, but not after abdominal surgery reduced intestinal inflammation and prevented POI in wild-type, but not in α7nAChR knockout mice. In humans, preoperative administration of prucalopride, but not of VNS, decreased Il6 and Il8 expression in the muscularis externa and improved clinical recovery.ConclusionEnteric neurons dampen mMφ activation, an effect mimicked by prucalopride. Preoperative, but not postoperative treatment with prucalopride prevents intestinal inflammation and shortens POI in both mice and human, indicating that preoperative administration of 5-HT4R agonists should be further evaluated as a treatment of POI.Trial registration numberNCT02425774.

scholarly journals Progesterone Receptor Isoforms A and B: Temporal and Spatial Differences in Expression during Murine Mammary Gland Development

Endocrinology ◽  
2005 ◽  
Vol 146 (8) ◽  
pp. 3577-3588 ◽  
Author(s):  
Mark D. Aupperlee ◽  
Kyle T. Smith ◽  
Anastasia Kariagina ◽  
Sandra Z. Haslam
Keyword(s):  
Mammary Gland ◽  
Progesterone Receptor ◽  
Cyclin D1 ◽  
Mammary Gland Development ◽  
Minor Role ◽  
Normal Mammary Gland ◽  
Temporal Separation ◽  
Stage Of Development ◽  
Gland Development

Abstract Progesterone is a potent mitogen in the mammary gland. Based on studies using cells and animals engineered to express progesterone receptor (PR) isoforms A or B, PRA and PRB are believed to have different functions. Using an immunohistochemical approach with antibodies specific for PRA only or PRB only, we show that PRA and PRB expression in mammary epithelial cells is temporally and spatially separated during normal mammary gland development in the BALB/c mouse. In the virgin mammary gland when ductal development is active, the only PR protein isoform expressed was PRA. PRA levels were significantly lower during pregnancy, suggesting a minor role at this stage of development. PRB was abundantly expressed only during pregnancy, during alveologenesis. PRA and PRB colocalization occurred in only a small percentage of cells. During pregnancy there was extensive colocalization of PRB with 5-bromo-2′-deoxyuridine (BrdU) and cyclin D1; 95% of BrdU-positive cells and 83% of cyclin D1-positive cells expressed PRB. No colocalization of PRA with either BrdU or cyclin D1 was observed at pregnancy. In the virgin gland, PRA colocalization with BrdU or cyclin D1 was low; only 27% of BrdU-positive cells and 4% of cyclin D1-positive cells expressed PRA. The implication of these findings is that different actions of progesterone are mediated in PRB positive vs. PRA-positive cells in vivo. The spatial and temporal separation of PR isoform expression in mouse mammary gland provides a unique opportunity to determine the specific functions of PRA vs. PRB in vivo.

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