Progesterone-dependent and -independent modulation of luminal epithelial transcription to support pregnancy in cattle

In cattle, starting 4-5 days after estrus, pre-implantation embryonic development occurs in the confinement of the uterine lumen. Cells in the endometrial epithelial layer control the molecular traffic to and from the lumen and, thereby determine luminal composition. Starting early post-estrus, endometrial function is regulated by sex-steroids, but the effects of progesterone on luminal cells transcription have not been measured in vivo. First objective was to determine the extent to which progesterone controls transcription in luminal epithelial cells 4 d (D4) after estrus. Second objective was to discover luminal transcripts that predict pregnancy outcomes, when the effect of progesterone is controlled. Endometrial luminal epithelial cells were collected from embryo transfer recipients on D4 using a cytological brush and their transcriptome determined by RNASeq. Pregnancy by embryo transfer was measured on D30 (25 pregnant and 18 non-pregnant). Progesterone concentration on D4 was associated positively (n= 182) and negatively (n= 58) with gene expression. Progesterone-modulated transcription indicated an increase in oxidative phosphorylation, biosynthetic activity and proliferation of epithelial cells. When these effects of progesterone were controlled, different genes affected positively (n= 22) and negatively (n= 292) odds of pregnancy. These set of genes indicated that a receptive uterine environment was characterized by the inhibition of phosphoinositide signaling and innate immune system responses. A panel of 25 genes predicted the pregnancy outcome with sensitivity and specificity ranging from 64-96% and 44-83%, respectively. In conclusion, in the early diestrus, both progesterone-dependent and -independent mechanisms regulate luminal epithelial transcription associated with pregnancy outcomes in cattle.

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Ultrastructural Contributions to the Histogenesis of Pleomorphic Adenoma

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053759 ◽

1982 ◽

Vol 40 ◽

pp. 218-219

Author(s):

I. Dardick ◽

A.W.P. van Nostrand ◽

Diane Jeans ◽

P. Rippstein ◽

V. Edwards

Keyword(s):

Epithelial Cells ◽

Tumor Cells ◽

Pleomorphic Adenoma ◽

Cell Types ◽

Myoepithelial Cells ◽

Luminal Type ◽

Cell Processes ◽

Luminal Cells ◽

Sick Children ◽

Luminal Epithelial Cells

Hospital, Ottawa, Canada and ^Hospital for Sick Children, Toronto, Canada. Survey-type electron micrographs correlated with semithin plastic sections (Fig. 2) were used in an ultrastructural study of 24 cases of salivary gland pleomorphic adenoma in order to assess tumor cell types and their organization in cellular regions and the gradual alterations occurring with the development of myxoid areas. Such micrographs confirm the presence of two principal cell types with smaller numbers of highly organized luminal epithelial cells forming duct- or acinar-like structures and more numerous, angular, mosaically or loosely arranged tumor cells surrounding luminal type cells. As is evident in Figure 1, darker staining, angular tumor cells just external to duct luminal cells have a specific and intimate association with luminal cells through cell processes and well developed desmosomes. Despite the lack of classical features of myoepithelial cells, the organizational arrangement of the two cell types and the distinctly different cytologic features of tumor cells external to luminal epithelial cells suggests that the former cell type represents myoepithelial cells modified as a result of neoplastic induction (Figs. 1 and 2).

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A subclass of luminal epithelial cells in the human mammary gland, defined by antibodies to cytokeratins

Journal of Cell Science ◽

10.1242/jcs.75.1.17 ◽

1985 ◽

Vol 75 (1) ◽

pp. 17-33

Author(s):

J. Bartek ◽

E.M. Durban ◽

R.C. Hallowes ◽

J. Taylor-Papadimitriou

Keyword(s):

Epithelial Cells ◽

Mammary Epithelial ◽

Immunofluorescent Staining ◽

Immunoperoxidase Staining ◽

Proliferative Potential ◽

Collagen Gels ◽

Tissue Sections ◽

Cell Extracts ◽

Luminal Cells ◽

Luminal Epithelial Cells

Two monoclonal antibodies, BA16 and BA17, have been developed using a detergent-insoluble extract of human mammary epithelial organoids as immunogen. Indirect immunofluorescent staining of cultured cells showed that the component reacting with the antibodies was filamentous and the intensity of staining was stronger in mitotic cells. Immunoblotting of cell extracts showed that both antibodies react with only one band of 40 X 10(3) molecular weight, which was present in keratin-enriched extracts of cells or organoids. Furthermore, the tissue distribution of the component reacting with the antibodies was that predicted for human keratin 19. The antibodies showed differences in the intensity of staining of cells or tissue sections fixed and prepared in different ways indicating that they reacted with different epitopes. The pattern of expression of the 40 X 10(3) Mr keratin by normal mammary epithelial cells was investigated by immunoperoxidase staining of tissue sections, cultured milk cells, and organoids of different sizes cultured in collagen gels. It was found that basal or myoepithelial cells did not express this keratin. Some heterogeneity of expression of this component was seen in luminal epithelial cells, found almost exclusively in the smaller structures. These cells did, however, express other keratins characteristic of luminal cells. The distribution in the mammary tree of the luminal cells that did not express the 40 X 10(3) Mr keratin appears to be similar to that expected for cells with the proliferative potential to produce new terminal ductal lobular units or an increase in branching of existing terminal ductal lobular units. It is shown that these cells have considerable proliferative potential by the fact that they form large colonies in milk cell cultures.

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Reporters to mark and eliminate basal or luminal epithelial cells in culture and in vivo

PLoS Biology ◽

10.1371/journal.pbio.2004049 ◽

2018 ◽

Vol 16 (6) ◽

pp. e2004049 ◽

Cited By ~ 5

Author(s):

Olmo Sonzogni ◽

Jennifer Haynes ◽

Laurie A. Seifried ◽

Yahia M. Kamel ◽

Kai Huang ◽

...

Keyword(s):

Epithelial Cells ◽

Cells In Culture ◽

Luminal Epithelial Cells

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Normal and tumor-derived myoepithelial cells differ in their ability to interact with luminal breast epithelial cells for polarity and basement membrane deposition

Journal of Cell Science ◽

10.1242/jcs.115.1.39 ◽

2002 ◽

Vol 115 (1) ◽

pp. 39-50 ◽

Cited By ~ 11

Author(s):

Thorarinn Gudjonsson ◽

Lone Rønnov-Jessen ◽

René Villadsen ◽

Fritz Rank ◽

Mina J. Bissell ◽

...

Keyword(s):

Breast Cancer ◽

Epithelial Cells ◽

Basement Membrane ◽

Specific Antigen ◽

Myoepithelial Cells ◽

Normal Breast ◽

Breast Epithelial ◽

Luminal Epithelial Cells ◽

Laminin 1

The signals that determine the correct polarity of breast epithelial structures in vivo are not understood. We have shown previously that luminal epithelial cells can be polarized when cultured within a reconstituted basement membrane gel. We reasoned that such cues in vivo may be given by myoepithelial cells. Accordingly, we used an assay where luminal epithelial cells are incorrectly polarized to test this hypothesis. We show that culturing human primary luminal epithelial cells within collagen-I gels leads to formation of structures with no lumina and with reverse polarity as judged by dual stainings for sialomucin, epithelial specific antigen or occludin. No basement membrane is deposited, and β4-integrin staining is negative. Addition of purified human myoepithelial cells isolated from normal glands corrects the inverse polarity, and leads to formation of double-layered acini with central lumina. Among the laminins present in the human breast basement membrane (laminin-1, -5 and -10/11), laminin-1 was unique in its ability to substitute for myoepithelial cells in polarity reversal.Myoepithelial cells were purified also from four different breast cancer sources including a biphasic cell line. Three out of four samples either totally lacked the ability to interact with luminal epithelial cells, or conveyed only correction of polarity in a fraction of acini. This behavior was directly related to the ability of the tumor myoepithelial cells to produce α-1 chain of laminin. In vivo, breast carcinomas were either negative for laminin-1 (7/12 biopsies) or showed a focal, fragmented deposition of a less intensely stained basement membrane (5/12 biopsies). Dual staining with myoepithelial markers revealed that tumor-associated myoepithelial cells were either negative or weakly positive for expression of laminin-1, establishing a strong correlation between loss of laminin-1 and breast cancer. We conclude that the double-layered breast acinus may be recapitulated in culture and that one reason for the ability of myoepithelial cells to induce polarity is because they are the only source of laminin-1 in the breast in vivo. A further conclusion is that a majority of tumor-derived/-associated myoepithelial cells are deficient in their ability to impart polarity because they have lost their ability to synthesize sufficient or functional laminin-1. These results have important implications for the role of myoepithelial cells in maintenance of polarity in normal breast and how they may function as structural tumor suppressors.

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PSIII-9 Supplementation of beta-carotene or fatty acids alters production of prostaglandins in bovine endometrial epithelial cells

Journal of Animal Science ◽

10.1093/jas/skab235.530 ◽

2021 ◽

Vol 99 (Supplement_3) ◽

pp. 288-289

Author(s):

Allison R Harman ◽

Rebecca Swanson

Keyword(s):

Epithelial Cells ◽

Antioxidant Properties ◽

Beta Carotene ◽

Endometrial Cells ◽

Prostaglandin Production ◽

Uterine Environment ◽

Free Media ◽

Endometrial Epithelial Cells

Abstract Differential prostaglandin secretion from the bovine endometrium can be used as a marker for an embryotropic or embryotoxic uterine environment. Beta-carotene has antioxidant properties and is the precursor for retinol, which has been shown to improve early embryonic development in vivo and in vitro. Furthermore, dietary fatty acid supplementation, specifically eicosapentaenoic acid (EPA) and decosahexaenoic acid (DHA) has been shown to alter prostaglandin production. The objective of this study was to determine prostaglandin production of endometrial cells following treatment with beta-carotene, EPA, or DHA. Bovine endometrial epithelial cells were treated for 24 hours with serum-free media supplemented with either 10 µM beta-carotene, 10 µM EPA, 10 µM DHA or ethanol (>1% volume/volume) vehicle control. After treatment, concentrations of PGE2 and PGF2a were analyzed in media via commercially available ELISA kits. Concentrations and ratios of prostaglandins were analyzed via ANOVA using the mixed procedure in SAS version 9.4. Beta-carotene treatment decreased PGE2 (P < 0.0001) and PGF2a (P = 0.0003) concentrations in media compared to controls. However, the ratio of PGE2:PGF2a was not different (P = 0.1203) between beta-carotene and controls. DHA treatment decreased PGE2 (P < 0.0001) concentrations in media but did not alter (P = 0.1079) PGF2a concentrations in media compared to controls. The ratio of PGE2:PGF2a was not different (P = 0.6343) between DHA and controls. EPA treatment did not alter (P = 0.1503) PGE2 concentrations in media compared to controls. Conversely, PGF2a concentrations were decreased (P = 0.0088) in media treated with EPA compared to controls. Therefore, the ratio of PGE2:PGF2a was increased (P = 0.0116) between EPA versus controls. These studies demonstrate that in vitro supplementation of EPA may alter the endometrial synthesis of prostaglandins to be more embryotropic. Therefore, EPA may be therapeutic for in vivo trials to influence the early uterine environment.

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Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

BioMed Research International ◽

10.1155/2020/7120375 ◽

2020 ◽

Vol 2020 ◽

pp. 1-15 ◽

Cited By ~ 1

Author(s):

Katarzyna Wojtanowicz-Markiewicz ◽

Magdalena Kulus ◽

Sandra Knap ◽

Ievgenia Kocherova ◽

Maurycy Jankowski ◽

...

Keyword(s):

Cell Culture ◽

Epithelial Cells ◽

Primary Cell Culture ◽

Water Movement ◽

Inorganic Ions ◽

Primary Cell ◽

Lag Phase ◽

Luminal Epithelial Cells

Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx’s behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.

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ΔN63 suppresses the ability of pregnancy-identified mammary epithelial cells (PIMECs) to drive HER2-positive breast cancer

Cell Death and Disease ◽

10.1038/s41419-021-03795-5 ◽

2021 ◽

Vol 12 (6) ◽

Author(s):

Christopher E. Eyermann ◽

Jinyu Li ◽

Evguenia M. Alexandrova

Keyword(s):

Breast Cancer ◽

Tumor Suppressor ◽

Epithelial Cells ◽

Egf Receptor ◽

Mammary Epithelial Cells ◽

Mammary Epithelial ◽

Self Renewal ◽

Luminal Cells ◽

Positive Breast Cancer

AbstractWhile pregnancy is known to reduce a woman’s life-long risk of breast cancer, clinical data suggest that it can specifically promote HER2 (human EGF receptor 2)-positive breast cancer subtype (HER2+ BC). HER2+ BC, characterized by amplification of HER2, comprises about 20% of all sporadic breast cancers and is more aggressive than hormone receptor-positive breast cancer (the majority of cases). Consistently with human data, pregnancy strongly promotes HER2+ BC in genetic mouse models. One proposed mechanism of this is post-pregnancy accumulation of PIMECs (pregnancy-identified mammary epithelial cells), tumor-initiating cells for HER2+ BC in mice. We previously showed that p63, a homologue of the tumor suppressor p53, is required to maintain the post-pregnancy number of PIMECs and thereby promotes HER2+ BC. Here we set to test whether p63 also affects the intrinsic tumorigenic properties of PIMECs. To this end, we FACS-sorted YFP-labeled PIMECs from p63+/−;ErbB2 and control p63+/+;ErbB2 females and injected their equal amounts into immunodeficient recipients. To our surprise, p63+/− PIMECs showed increased, rather than decreased, tumorigenic capacity in vivo, i.e., significantly accelerated tumor onset and tumor growth, as well as increased self-renewal in mammosphere assays and proliferation in vitro and in vivo. The underlying mechanism of these phenotypes seems to be a specific reduction of the tumor suppressor TAp63 isoform in p63+/− luminal cells, including PIMECs, with concomitant aberrant upregulation of the oncogenic ΔNp63 isoform, as determined by qRT-PCR and scRNA-seq analyses. In addition, scRNA-seq revealed upregulation of several cancer-associated (Il-4/Il-13, Hsf1/HSP), oncogenic (TGFβ, NGF, FGF, MAPK) and self-renewal (Wnt, Notch) pathways in p63+/−;ErbB2 luminal cells and PIMECs per se. Altogether, these data reveal a complex role of p63 in PIMECs and pregnancy-associated HER2+ BC: maintaining the amount of PIMECs while suppressing their intrinsic tumorigenic capacity.

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Prokineticin 1–prokineticin receptor 1 signaling promotes angiogenesis in the porcine endometrium during pregnancy†

Biology of Reproduction ◽

10.1093/biolre/ioaa066 ◽

2020 ◽

Vol 103 (3) ◽

pp. 654-668 ◽

Cited By ~ 1

Author(s):

Ewelina Goryszewska ◽

Piotr Kaczynski ◽

Gianfranco Balboni ◽

Agnieszka Waclawik

Keyword(s):

Epithelial Cells ◽

Reproductive Tract ◽

Secretory Protein ◽

Angiogenic Factor ◽

In Vivo Model ◽

Secretory Function ◽

Pleiotropic Functions ◽

Estradiol 17Β ◽

Luminal Epithelial Cells

Abstract Pregnancy establishment in mammals, including pigs, requires proper communication between embryos and the maternal reproductive tract. Prokineticin 1 (PROK1) has been described as a secretory protein with pleiotropic functions and as a novel tissue-specific angiogenic factor. However, despite the studies performed mainly on human cell lines and in mice, the function of PROK1 in the endometrium during early pregnancy is still not fully elucidated. We hypothesized that PROK1 contributes to pregnancy establishment in pigs. The present study is the first to report that the expression of PROK1 and its receptor (PROKR1) is elevated in the porcine endometrium during the implantation and early placentation period. PROK1 protein was detected mainly in luminal epithelial cells, glandular epithelial cells, and blood vessels in the endometrium. Using the porcine in vivo model of unilateral pregnancy, we revealed that conceptuses induced the endometrial expression of PROK1 and PROKR1. Moreover, the embryonic signal, estradiol-17β, as well as progesterone, stimulated the endometrial expression of PROK1 and PROKR1. We also evidenced that PROK1–PROKR1 signaling supports endometrial angiogenesis in pigs. The PROK1-stimulated proliferation of primary porcine endometrial endothelial (PEE) cells involved PI3K/AKT/mTOR, MAPK, cAMP, and NFKB signaling pathways. Furthermore, PROK1 via PROKR1 promoted the formation of capillary-like structures by PEE cells. PROK1 also stimulated VEGFA and PGF2α secretion, which in turn may indirectly support angiogenic changes within endometrial tissue. In summary, our study suggests that PROK1 acts as an embryonic signal mediator that regulates endometrial angiogenesis and secretory function during the implantation and early placentation period in pigs.

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Anti-estrogenic effect of TAS-108 in breast cancer cell lines.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.e11029 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. e11029-e11029

Author(s):

Hadong Kim ◽

Young Choi ◽

Tae-Hoon Kim

Keyword(s):

Breast Cancer ◽

Epithelial Cells ◽

Growth Inhibition ◽

Cell Lines ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Breast Cancer Cell Lines ◽

Luminal Epithelial Cells ◽

Mcf 7

e11029 Background: TAS-108 (TAS) is an antiestrogen that binds to both ERα and ERβ and inhibits both estrogen-dependent and independent tumors and TAM resistant tumors. Breast cancer cell lines (BCC) with different pheno- and genotypes are reasonably acceptable models of in vivo breast cancer cell behavior. Thus, we have investigated anti-estrogenic activity of TAS on BCC in the presence of diarylpropionitrile (DPN), a pure ERβ agonist to find ERβ role in TAS activity. Methods: Cultured cells of (1) MCF-7 and ZR-75 (luminal epithelial cells), (2) BT-474 and SK-BR-3 (weakly luminal epithelial cells), and (3) MDA-MB-231 and Hs578T (invasive, mesenchymal-like cells) were treated with: (1) increasing concentrations of DPN or E2, (2) 100nM of 4-hydroxytamoxifen (4-HT) or TAS, and (3) 100nM of DPN or E2 with increasing concentrations of either 4-HT or TAS. After 6 days, cell proliferation was analyzed. The mRNA levels of ERβ and ERα isoform were determined by RT-qPCR. ERα, PR, and Her-2 status was determined by IHC. Percent proliferation compared to the control was calculated and P <0.05 based on T-test was considered significant. Results: The most significant proliferation and growth inhibition was seen in MCF-7 BCC. In the presence of DPN, IC50 (50% inhibitory concentration) was 12.5nmol of TAS vs 33.1nmol of 4-HT. In the presence of E2, IC50 was 2.15 nmol of TAS vs 49.7nmol of 4-HT. Other BCC showed insignificant or variable proliferative and inhibitory response. Overall, DPN displayed high proliferation in most BCC. TAS exhibited higher growth inhibition than 4-HT. Conclusions: Variable responses of different BCC lines to ERα and ERβ agonists and antagonists may be instructive in modeling in vivo breast cancer response to these compounds. Overall, exposure to ERβ agonists showed a significant proliferation in ERα+, ERα- and ERβ+ cells. TAS activity was more pronounced and its anti-estrogen activity may involve ERβ in addition to ERα. [Table: see text]

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Sperm migration, selection, survival and fertilizing ability in the mammalian oviduct

Biology of Reproduction ◽

10.1093/biolre/ioab105 ◽

2021 ◽

Author(s):

Coline Mahé ◽

Aleksandra Maria Zlotkowska ◽

Karine Reynaud ◽

Guillaume Tsikis ◽

Pascal Mermillod ◽

...

Keyword(s):

Epithelial Cells ◽

Mammalian Species ◽

Female Reproductive Tract ◽

Vitro Fertilization ◽

Domestic Species ◽

Fertilizing Ability ◽

Sperm Migration ◽

Luminal Epithelial Cells

Abstract In vitro fertilization (IVF) gives rise to embryos in a number of mammalian species and is currently widely used for assisted reproduction in humans and for genetic purposes in cattle. However, the rate of polyspermy is generally higher in vitro than in vivo and IVF remains ineffective in some domestic species like pigs and horses, highlighting the importance of the female reproductive tract for gamete quality and fertilization. In this review, the way the female environment modulates sperm selective migration, survival and acquisition of fertilizing ability in the oviduct is being considered under six aspects: (1) the utero-tubal junction which selects a sperm sub-population entering the oviduct; (2) the presence of sperm binding sites on luminal epithelial cells in the oviduct, which prolong sperm viability and plays a role in limiting polyspermic fertilization; (3) the contractions of the oviduct, which promote sperm migration toward the site of fertilization in the ampulla; (4) the regions of the oviduct, which play different roles in regulating sperm physiology and interactions with oviduct epithelial cells; (5) the time of ovulation and (6) the steroid hormonal environment which regulates sperm release from the luminal epithelial cells and facilitates capacitation in a finely orchestrated manner.

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