Ciliated epithelial-specific and regional-specific expression and regulation of the estrogen receptor-β2 in the fallopian tubes of immature rats: a possible mechanism for estrogen-mediated transport process in vivo

2007 ◽  
Vol 293 (1) ◽  
pp. E147-E158 ◽  
Author(s):  
Ruijin Shao ◽  
Birgitta Weijdegård ◽  
Julia Fernandez-Rodriguez ◽  
Emil Egecioglu ◽  
Changlian Zhu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Transport Process ◽  
Intracellular Localization ◽  
Confocal Imaging ◽  
Dependent Manner ◽  
Fallopian Tubes ◽  
Specific Expression ◽  
Ample Evidence ◽  
Ciliated Epithelial Cells

Several ERβ isoforms have been identified in human and rodent tissues, but it is unclear whether each isoform has distinctly different cellular targeting characteristics and physiological functions. We have investigated the intracellular localization and regulatory patterns for ERβ isoforms in rat fallopian tubes. Western blot analysis reveals that two ERβ isoforms corresponding to ERβ1 and ERβ2 are expressed in rat fallopian tubes. However, ERβ2 is the predominant form of ERβ in this tissue. High-resolution confocal imaging and immunohistochemical analysis provide ample evidence that ERβ expression is limited almost exclusively to the ciliated epithelial cells, in contrast to ERα, which is widely distributed. Furthermore, within the ciliated epithelial cells, ERβ is colocalized with β-tubulin IV at stem portion of the cilia. We show that ERβ2 protein expression is tightly regulated by E2 or DPN in a time-dependent manner without changes in ERβ1 expression. These estrogenic effects are inhibited by an ER antagonist, ICI 182,780. In addition, significant alteration of ERβ immunoreactivity is detected only histologically in the ampullary region. Since the cilia are considered an essential determinant of tubal transport, we further demonstrate that E2- or DPN-induced ERβ2 activation is associated with alterations in tubal protein expression crucial for the regulation of calcium-dependent ciliary beating. Given the coordinated regulation and interaction of ER and progesterone receptor in the cilia, we hypothesize that tubal ERβ2 may facilitate the estrogen-mediated transport process by processing protein-protein interaction under physiological and/or pathological conditions. We show for the first time that a previously unrecognized localization of ERβ isoform in rat fallopian tubes can combine with estrogen to individually control the expression of ER β-isoforms in normal target tissues.

scholarly journals Interferon-γ inhibits sirtuin 6 gene expression in intestinal epithelial cells through a microRNA-92b-dependent mechanism

2020 ◽  
Vol 318 (4) ◽  
pp. C732-C739
Author(s):  
Fangyi Liu ◽  
Xiao Wang ◽  
Hua Geng ◽  
Heng-Fu Bu ◽  
Peng Wang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Intestinal Epithelial Cells ◽  
Specific Inhibitor ◽  
In Silico Analysis ◽  
Interferon Γ ◽  
Luciferase Assay ◽  
Epithelial Cell Line ◽  
Dependent Manner ◽  
Intestinal Epithelial

Sirtuin 6 (Sirt6) is predominantly expressed in epithelial cells in intestinal crypts. It plays an important role in protecting intestinal epithelial cells against inflammatory injury. Previously, we found that colitis is associated with the downregulation of Sirt6 protein in the intestines. Here, we report that murine interferon-γ (Ifnγ) inhibits Sirt6 protein but not mRNA expression in young adult mouse colonocytes (YAMC, a mouse colonic epithelial cell line) in a dose- and time-dependent manner. Using microRNA array analysis, we showed that Ifnγ induces expression of miR-92b in YAMC cells. With in silico analysis, we found that the Sirt6 3′-untranslated region (UTR) contains a putative binding site for miR-92b. Luciferase assay showed that Ifnγ inhibited Sirt6 3′-UTR activity and this effect was mimicked by miR-92b via directly targeting the miR-92b seed site in the 3′-UTR of Sirt6 mRNA. Furthermore, Western blot demonstrated that miR-92b downregulated Sirt6 protein expression in YAMC cells. Blocking miR-92b with a specific inhibitor attenuated the inhibitory effect of Ifnγ on Sirt6 protein expression in the cells. Collectively, our data suggest that Ifnγ inhibits Sirt6 protein expression in intestinal epithelial cells via a miR-92b-mediated mechanism. miR-92b may be a novel therapeutic target for rescuing Sirt6 protein levels in intestinal epithelial cells, thereby protecting against intestinal mucosal injury caused by inflammation.

scholarly journals Ovarian steroids affect prostaglandin production in equine endometrial cells in vitro

2014 ◽  
Vol 220 (3) ◽  
pp. 263-276 ◽  
Author(s):  
Anna Z Szóstek ◽  
António M Galvão ◽  
Graça M Ferreira-Dias ◽  
Dariusz J Skarzynski
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cells ◽  
Protein Expression ◽  
Stromal Cells ◽  
Positive Control ◽  
Dependent Manner ◽  
Ovarian Steroids ◽  
Endometrial Cells ◽  
Prostaglandin Endoperoxide Synthase

This study aimed to evaluate the influence of ovarian steroids on equine endometrial epithelial and stromal cells, specifically i) prostaglandin (PG) production in a time-dependent manner, ii) specific PG synthases mRNA transcription and protein expression, and iii) cell proliferation. After passage I, cells were exposed to vehicle, oxytocin (OT, positive control, 10−7M), progesterone (P4, 10−7M), 17β estradiol (E2, 10−9M), or P4+E2for 12, 24, 48, or 72 h. Following treatment, PG concentration was determined using the direct enzyme immunoassay (EIA) method. Alterations inPGsynthases mRNA transcriptions,PGsynthases protein expression, and cell proliferation in response to the treatments were determined after 24 h using real-time PCR, western blot, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide respectively. After 24 h, E2and P4+E2increased PGE2and PGF2αsecretion as well as specific prostaglandin-endoperoxide synthase-2 (PTGS2), PGE2synthases (PGES), and PGF2αsynthases (PGFS) expression in the epithelial cells (P<0.05). Additionally, E2and P4+E2increased PTGS2 expression in stromal cells after 24 h (P<0.05). In stromal cells, P4+E2increased PGE2production as well as PGES expression after 24 h (P<0.05). Both E2and P4+E2increased PGF2αproduction by stromal cells after 24 h (P<0.05). Ovarian steroids affected proliferation of stromal and epithelial cells during the 24-h incubation period (P<0.05). We provide evidence that ovarian steroids affect PG production in equine endometrial cells, upregulating PTGS2, PGES, and PGFS expression. Ovarian steroid-stimulated PG production could be an important mechanism occurring in the equine endometrium that is involved in the regulation of the estrous cycle and early pregnancy.

scholarly journals Milk-derived exosomes (MDEs) have a different biological effect on normal fetal colon epithelial cells compared to colon tumor cells in a miRNA-dependent manner

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Shimon Reif ◽  
Yaffa Elbaum Shiff ◽  
Regina Golan-Gerstl
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Human Milk ◽  
Tumor Cells ◽  
Expression Profiles ◽  
Biological Effects ◽  
Biologically Active ◽  
Type I ◽  
Dependent Manner ◽  
Mesenchymal Transformation

Abstract Background Breastfeeding is the ideal source of infant nutrition. Human milk consists not only of nutrients but also biologically active components. Among these latter compounds, exosomes contain proteins, lipids, mRNAs and miRNAs. Methods To elucidate the biological effects of milk-derived exosomes (MDEs) on normal colonic epithelial cells compared to colonic tumor cells, we incubated cells with MDEs. MDEs were able to enter into normal and tumor cells and change their miRNA expression profiles. Proliferation, cell morphology and protein expression were analyzed in these cells. Results Human milk-derived exosomes induced proliferation- and epithelial mesenchymal transformation-related changes, such as collagen type I and twist expression, in normal but not in tumor cells. PTEN, a target of miRNA-148a, was downregulated in normal but not in tumor cells following incubation with MDEs. Moreover, miRNA-148a-3p knockdown cells were used to demonstrate the importance of miRNA in the effect of exosomes on cell proliferation and protein expression. MDEs inhibited proliferation and DNMT1 expression in cells with knockdown of miRNA-148a. Conclusions In conclusion, the positive effect of exosomes on normal cells without affecting tumor cells may presents an aspect of their safety when considering it use as a nutritional supplement to infant formula.

Downregulation of cilia-localized Il-6Rα by 17β-estradiol in mouse and human fallopian tubes

2009 ◽  
Vol 297 (1) ◽  
pp. C140-C151 ◽  
Author(s):  
Ruijin Shao ◽  
Magdalena Nutu ◽  
Linda Karlsson-Lindahl ◽  
Anna Benrick ◽  
Birgitta Weijdegård ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Fallopian Tube ◽  
Knockout Mice ◽  
Cellular Localization ◽  
Molecular Mechanisms ◽  
Fallopian Tubes ◽  
Female Reproduction ◽  
Specific Expression ◽  
17Β Estradiol ◽  
Specific Regulation

The action of interleukin-6 (IL-6) impacts female reproduction. Although IL-6 was recently shown to inhibit cilia activity in human fallopian tubes in vitro, the molecular mechanisms underlying IL-6 signaling to tubal function remain elusive. Here, we investigate the cellular localization, regulation, and possible function of two IL-6 receptors (IL-6Rα and gp130) in mouse and human fallopian tubes in vivo. We show that IL-6Rα is restricted to the cilia of epithelial cells in both mouse and human fallopian tubes. Exogenous 17β-estradiol (E2), but not progesterone (P4), causes a time-dependent decrease in IL-6Rα expression, which is blocked by the estrogen receptor (ER) antagonist ICI-182,780. Exposure of different ER-selective agonists propyl-(1H)-pyrazole-1,3,5-triyl-trisphenol or 2,3-bis-(4-hydroxyphenyl)-propionitrile demonstrated an ER subtype-specific regulation of IL-6Rα in mouse fallopian tubes. In contrast to IL-6Rα, gp130 was detected in tubal epithelial cells in mice but not in humans. In humans, gp130 was found in the muscle cells and was decreased in the periovulatory and luteal phases during the reproductive cycles, indicating a species-specific expression and regulation of gp130 in the fallopian tube. Expression of tubal IL-6Rα and gp130 in IL-6 knockout mice was found to be normal; however, E2 treatment increased IL-6Rα, but not gp130, in IL-6 knockout mice when compared with wild-type mice. Furthermore, expression levels of IL-6Rα, but not gp130, decreased in parallel with estrogenic accelerated oocyte-cumulus complex (OCC) transport in mouse fallopian tubes. Our findings open the posibility that cilia-specific IL-6Rα may play a role in the regulation of OCC transport and suggest an estrogen-regulatory pathway of IL-6Rα in the fallopian tube.

Dopamine-induced translocation of protein kinase C isoforms visualized in renal epithelial cells

2000 ◽  
Vol 279 (6) ◽  
pp. C1812-C1818 ◽  
Author(s):  
Susana Nowicki ◽  
Maria Sol Kruse ◽  
Hjalmar Brismar ◽  
Anita Aperia
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Epithelial Cells ◽  
Subcellular Fractionation ◽  
Confocal Imaging ◽  
Receptor Subtype ◽  
Dependent Manner ◽  
Renal Epithelial Cells ◽  
Kinase C ◽  
Pkc Isoforms

Short-term regulation of sodium metabolism is dependent on the modulation of the activity of sodium transporters by first and second messengers. In understanding diseases associated with sodium retention, it is necessary to identify the coupling between these messengers. We have examined whether dopamine, an important first messenger in tubular cells, activates and translocates various protein kinase C (PKC) isoforms. We used a proximal tubular-like cell line, LLCPK-1 cells, in which dopamine was found to inhibit Na+-K+-ATPase in a PKC-dependent manner. Translocation of PKC isoforms was studied with both subcellular fractionation and confocal microscopy. Both techniques revealed a dopamine-induced translocation from cytosol to plasma membrane of PKC-α and -ε, but not of PKC-δ, -γ, and -ζ. The process of subcellular fractionation resulted in partial translocation of PKC-ε. This artifact was eliminated in confocal studies. Confocal imaging permitted detection of translocation within 20 s. Translocation was abolished by a phospholipase C inhibitor and by an antagonist against the dopamine 1 subtype (D1) but not the 2 subtype of receptor (D2). In conclusion, this study visualizes in renal epithelial cells a very rapid activation of the PKC-α and -ε isoforms by the D1 receptor subtype.

scholarly journals Cloning and subcellular localization of novel rab proteins reveals polarized and cell type-specific expression

1994 ◽  
Vol 107 (12) ◽  
pp. 3437-3448 ◽  
Author(s):  
A. Lutcke ◽  
R.G. Parton ◽  
C. Murphy ◽  
V.M. Olkkonen ◽  
P. Dupree ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cultured Cells ◽  
Intracellular Localization ◽  
Small Gtpases ◽  
Apical Plasma Membrane ◽  
Rab Proteins ◽  
Dependent Manner ◽  
Cell Type ◽  
Apical Side ◽  
Polarized Cells

Small GTPases of the rab subfamily are specific regulators of vesicular transport. The intracellular localization of these proteins has been mostly investigated in cultured cells where they have been found associated with distinct compartments of the exocytic and endocytic pathways. Using a PCR-based cloning approach we have recently identified several novel rab proteins, extending the total number of this family to more than 30 members. Here, we have investigated the mRNA expression in different tissues and the intracellular localization in organ cryosections of two rab proteins, rab18 and rab20. Both northern blot analysis and confocal immunofluorescence microscopy demonstrated that these proteins are expressed in a tissue- and cell type-dependent manner. Despite their presence in non-polarized cells and polarized cells, both proteins are highly expressed on the apical side of kidney tubule epithelial cells. Electron microscopic studies revealed that rab18 and rab20 are located in apical dense tubules, endocytic structures underlying the apical plasma membrane, suggesting that they play a role in apical endocytosis/recycling. In intestinal epithelial cells as well, both proteins were localized apically, but, in addition, rab18 was found associated with the basolateral domain, suggesting that this protein is not restricted to the apical transport machinery of polarized epithelial cells. The results demonstrate that, depending on the epithelial cell type, rab proteins that are also expressed in non-polarized cells may be enriched in one or both surface domains. Together with the observed tissue- and cell type-dependent variation in the expression of the rab proteins, this suggests that the large number of mammalian rab proteins might reflect the specific requirements in the organization of membrane traffic encountered by different cell types.

Dynamic regulation of estrogen receptor-α isoform expression in the mouse fallopian tube: mechanistic insight into estrogen-dependent production and secretion of insulin-like growth factors

2007 ◽  
Vol 293 (5) ◽  
pp. E1430-E1442 ◽  
Author(s):  
Ruijin Shao ◽  
Emil Egecioglu ◽  
Birgitta Weijdegård ◽  
John J. Kopchick ◽  
Julia Fernandez-Rodriguez ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Fallopian Tube ◽  
Growth Hormone Receptor ◽  
Dependent Manner ◽  
Fallopian Tubes ◽  
Long Term Effects ◽  
Ici 182,780 ◽  
Isoform Expression ◽  
Insight Into

Estrogen receptors (ERs) are members of the nuclear receptor superfamily and are involved in regulation of fallopian tube functions (i.e., enhancement of protein secretion, formation of tubal fluid, and regulation of gamete transport). However, the ER subtype-mediated mechanisms underlying these processes have not been completely clarified. Recently, we identified ERβ expression and localization in rat fallopian tubes, suggesting a potential biological function of ERβ related to calcium-dependent ciliated beating. Here we provide for the first time insight into the less studied ERα isoforms, which mediate estrogen-dependent production and secretion of IGFs in vivo. First, Western blot studies revealed that three ERα isoforms were expressed in mouse fallopian tubes. Subsequent immunohistochemical analysis showed that ERα was detected in all cell types, whereas ERβ was mainly localized in ciliated epithelial cells. Second, ERα isoform levels were dramatically downregulated in mouse fallopian tubes by treatment with E2 or PPT, an ERα agonist, in a time-dependent manner. Third, the presence of ICI 182,780, an ER antagonist, blocked the E2- or PPT-induced downregulation of tubal ERα isoform expression in mice. However, alteration of ERα immunoreactivity following ICI 182,780 treatment was only detected in epithelial cells of the ampullary region. Fourth, changes in ERα isoform expression were found to be coupled to multiple E2 effects on tubal growth, protein synthesis, and secretion in mouse fallopian tube tissues and fluid. In particular, E2 exhibited positive regulation of IGF-I and IGF-II protein levels. Finally, using growth hormone receptor (GHR) gene-disrupted mice, we showed that regulation by E2 of IGF production was independent of GH-induced GHR signaling in mouse fallopian tubes in vivo. These data, together with previous studies from our laboratory, suggest that the long-term effects of estrogen agonist promote IGF synthesis and secretion in mouse tubal epithelial cells and fallopian tube fluid via stimulation of ERα.

scholarly journals Protein expression pattern of calcium-responsive transactivator in early postnatal and adult testes

2021 ◽  
Author(s):  
Ana Du ◽  
Li Li ◽  
Zhaoshuang Jiao ◽  
Gaochun Zhu ◽  
Ting Peng ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Expression Pattern ◽  
Sertoli Cells ◽  
Immunohistochemical Analysis ◽  
The Body ◽  
Spermatogenic Cells ◽  
Specific Expression ◽  
Adult Rat ◽  
Regulation Of Cell Cycle

AbstractCalcium-responsive transactivator (CREST), a nuclear protein highly expressed in postmitotic neurons, is involved in the regulation of cell cycle, differentiation and dendritic development of neuronal cells. Its mRNA has been detected in the testis of adult rat, whilst its protein expression and distribution pattern in the testis remain to be elucidated. In this study, we examined the distribution of CREST in the adult testes of both rats and human as well as the expression pattern of CREST in the testes of postnatal developing rats. In the adult testes of both human and rats, immunohistochemical analysis revealed that CREST was selectively distributed in the mature Sertoli cells but not in the spermatogenic cells. In the testes of postnatal developmental rats, CREST was expressed not only in Sertoli cells but also in the gonocytes and spermatogenic cells at the initial stage of spermatogenic cell differentiation. CREST immunoreactivity continued to increase in Sertoli cells during differentiation, reaching its peak in adulthood. However, CREST immunostaining intensity dramatically decreased as the spermatogenic cells differentiate, disappearing in the post-differentiation stage. Furthermore, Brg1 and p300, two CREST-interacting proteins ubiquitously expressed in the body, are found to be colocalized with CREST in the spermatogenic epithelial cells including Sertoli cells. The unique expression pattern of CREST in developing testis suggests that CREST might play regulatory roles in the differentiation of spermatogenic epithelial cells. The Sertoli cell-specific expression of CREST in the adulthood hints that CREST might be a novel biomarker for the mature Sertoli cells.

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