Abstract
The non-receptive uterine luminal epithelium forms a polarised epithelial barrier, protective against potential pathogenic assault from the external environment and invasion by the blastocyst. However, during the window of implantation, the uterine luminal epithelial cells (UECs) transition to a receptive state by dismantling many of their intercellular and cell–matrix adhesions in preparation for epithelial detachment and subsequent blastocyst implantation. The present study investigated the presence and regulation of the intercellular adhesion protein, Epithelial Cell Adhesion Molecule (EpCAM) during early pregnancy in the rat to understand its role in the transition to receptivity. Immunofluorescence and western blotting analysis were used to study EpCAM expression in normal pregnancy, hormone replacement studies and pseudopregnancy. EpCAM was abundantly expressed and localised to the uterine luminal and glandular epithelium during the non-receptive state but decreased to lower but still observable levels around the time of implantation. This decrease was not dependent on ovarian hormones or the blastocyst. Further, EpCAM colocalised with but did not associate with its frequent binding partner, Tumour necrosis factor α (TNFα)-converting enzyme, also known as A Disintegrin And Metalloprotease 17 (TACE/ADAM17), at the time of fertilisation. These results suggest that, prior to implantation, EpCAM mediates intercellular adhesion in the uterine epithelium, but that, during implantation when UECs lose the majority of their intercellular and cell–matrix adhesions, EpCAM levels are decreased but still present for the maintenance of mucosal integrity.
Similar content being viewed by others
References
Abrahamsohn P, Zorn T (1993) Implantation and decidualization in rodents. J Exp Zool 266:603–628
Aplin JD, Kimber SJ (2004) Trophoblast-uterine interactions at implantation. Reprod Biol Endocrinol 2:48–48
Balzar M, Bakker HA, Briaire-de-Bruijn IH et al (1998) Cytoplasmic tail regulates the intercellular adhesion function of the epithelial cell adhesion molecule. Mol Cell Biol 18:4833–4843
Balzar M, Winter MJ, de Boer CJ, Litvinov SV (1999) The biology of the 17-1A antigen (Ep-CAM). J Mol Med Berl 77:699–712
Cirulli VV, Crisa LL, Beattie GMG et al (1998) KSA antigen Ep-CAM mediates cell-cell adhesion of pancreatic epithelial cells: morphoregulatory roles in pancreatic islet development. J Cell Biol 140:1519–1534
de Boer CJ, van Krieken JH, Janssen-van Rhijn CM, Litvinov SV (1999) Expression of Ep-CAM in normal, regenerating, metaplastic, and neoplastic liver. J Pathol 188:201–206
Denker H-W (1993) Endometrial receptivity: cell biological aspects of an unusual epithelium. A review. Ann Anat 176:53–60
Denzel S, Maetzel D, Mack B et al (2008) Initial activation of EpCAM cleavage via cell-to-cell contact. BMC Cancer 9:402–402
Denzel S, Mack B, Eggert C et al (2012) MMP7 is a target of the tumour-associated antigen EpCAM. Int J Exp Pathol 93:341–353
Driemel C, Kremling H, Schumacher S et al (2013) Context-dependent adaption of EpCAM expression in early systemic esophageal cancer. Oncogene. doi:10.1038/onc.2013.441
Frederick BA, Helfrich BA, Coldren CD et al (2007) Epithelial to mesenchymal transition predicts gefitinib resistance in cell lines of head and neck squamous cell carcinoma and non-small cell lung carcinoma. Mol Cancer Ther 6:1683–1691
Gadalla S-E, Ojemalm K, Vasquez PL et al (2013) EpCAM associates with endoplasmic reticulum aminopeptidase 2 (ERAP2) in breast cancer cells. Biochem Biophys Res Commun 439:203–208
Gires O (2012) EpCAM in hepatocytes and their progenitors. J Hepatol 56:490–492
Goulet OO, Salomon JJ, Ruemmele FF et al (2006) Intestinal epithelial dysplasia (tufting enteropathy). Orphanet J Rare Dis 2:20–20
Groothuis PG, Dassen HHNM, Romano A, Punyadeera C (2007) Estrogen and the endometrium: lessons learned from gene expression profiling in rodents and human. Hum Reprod Update 13:405–417
Hussenet F, Dousset B, Cordonnier JL et al (1993) Tumour necrosis factor alpha and interleukin 2 in normal and infected human seminal fluid. Hum Reprod 8:409–411
Kaneko YY, Lindsay LAL, Murphy CRC (2008) Focal adhesions disassemble during early pregnancy in rat uterine epithelial cells. Reprod Fertil Dev 20:892–899
Kaneko Y, Lecce L, Day ML, Murphy CR (2011) β1 and β3 integrins disassemble from basal focal adhesions and β3 integrin is later localised to the apical plasma membrane of rat uterine luminal epithelial cells at the time of implantation. Reprod Fertil Dev 23:481–495
Kim J, Kang SG, Kim JI et al (2006) Implication of ADAM-8, −9, −10, −12, −15, −17, and ADAMTS-1 in implantational remodeling of a mouse uterus. Yonsei Med J 47:558–567
Ladwein M, Pape U-F, Schmidt D-S et al (2005) The cell-cell adhesion molecule EpCAM interacts directly with the tight junction protein claudin-7. Exp Cell Res 309:345–357
Le Naour F, Zöller M (2008) The tumor antigen EpCAM: tetraspanins and the tight junction protein claudin-7, new partners, new functions. Front Biosci 13:5847–5865. doi:10.2741/3121
Lei Z, Maeda T, Tamura A et al (2012) EpCAM contributes to formation of functional tight junction in the intestinal epithelium by recruiting claudin proteins. Dev Biol 371:136–145
Litvinov SV, Velders MP, Bakker HA et al (1994) Ep-CAM: a human epithelial antigen is a homophilic cell-cell adhesion molecule. J Cell Biol 125:437–446
Ljungkvist I (1971a) Attachment reaction of rat uterine luminal epithelium. II. The effect of progesterone on the morphology of the uterine glands and the luminal epithelium of the spayed, virgin rat. Acta Soc Med Ups 76:110–126
Ljungkvist I (1971b) Attachment reaction of rat uterine luminal epithelium. 3. The effect of estradiol, estrone and estriol on the morphology of the luminal epithelium of the spayed, virgin rat. Acta Soc Med Ups 76:139–157
Long JA and Evans HM (1922) The oestrous cycle in the rat and its associated phenomena. Memoirs of University of California 6:1–148
Lu DPD, Tian LL, O’Neill CC, King NJCN (2002) Regulation of cellular adhesion molecule expression in murine oocytes, peri-implantation and post-implantation embryos. Cell Res 12:373–383
Maetzel D, Denzel S, Mack B et al (2009) Nuclear signalling by tumour-associated antigen EpCAM. Nat Cell Biol 11:162–171
Maretzky T, Scholz F, Köten B et al (2008) ADAM10-mediated E-cadherin release is regulated by proinflammatory cytokines and modulates keratinocyte cohesion in eczematous dermatitis. J Invest Dermatol 128:1737–1746
Mueller JL, McGeough MD, Pena CA, Sivagnanam M (2014) Functional consequences of EpCAM mutation in mice and men. Am J Physiol Gastrointest Liver Physiol 306:G278–G288
Munz M, Kieu C, Mack B et al (2004) The carcinoma-associated antigen EpCAM upregulates c-myc and induces cell proliferation. Oncogene 23:5748–5758
Munz M, Zeidler R, Gires O (2005) The tumour-associated antigen EpCAM upregulates the fatty acid binding protein E-FABP. Cancer Lett 225:151–157
Murphy CR (2004) Uterine receptivity and the plasma membrane transformation. Cell Res 14:259–267
Nicholson MDO, Lindsay LA, Murphy CR (2010) Ovarian hormones control the changing expression of claudins and occludin in rat uterine epithelial cells during early pregnancy. Acta Histochem 112:42–52
Osta WA, Chen Y, Mikhitarian K et al (2004) EpCAM is overexpressed in breast cancer and is a potential target for breast cancer gene therapy. Cancer Res 64:5818–5824
Poon CE, Madawala RJ, Day ML, Murphy CR (2013) Claudin 7 is reduced in uterine epithelial cells during early pregnancy in the rat. Histochem Cell Biol 139:583–593
Schmidt D-SD, Klingbeil PP, Schnölzer MM, Zöller MM (2004) CD44 variant isoforms associate with tetraspanins and EpCAM. Exp Cell Res 297:329–347
Schnell U, Kuipers J, Giepmans BNG (2013) EpCAM proteolysis: new fragments with distinct functions? Biosci Rep 33:e00030. doi:10.1042/BSR20120128
Sharkey DJ, Tremellen KP, Jasper MJ et al (2012) Seminal fluid induces leukocyte recruitment and cytokine and chemokine mRNA expression in the human cervix after coitus. J Immunol 188:2445–2454. doi:10.4049/jimmunol.1102736
Sivagnanam M, Mueller JL, Lee H et al (2008) Identification of EpCAM as the gene for congenital tufting enteropathy. Gastroenterology 135:429–437
Trzpis MM, McLaughlin PMJP, de Lou MFHLM, Leij HMCM (2007) Epithelial cell adhesion molecule: more than a carcinoma marker and adhesion molecule. Am J Pathol 171:386–395
Wira CR, Grant-Tschudy KS, Crane-Godreau MA (2005) Epithelial cells in the female reproductive tract: a central role as sentinels of immune protection. Am J Reprod Immunol 53:65–76
Wu C-J, Mannan P, Lu M, Udey MC (2013) Epithelial cell adhesion molecule (EpCAM) regulates claudin dynamics and tight junctions. J Biol Chem 288:12253–12268
Yoon S-M, Gerasimidou D, Kuwahara R et al (2011) Epithelial cell adhesion molecule (EpCAM) marks hepatocytes newly derived from stem/progenitor cells in humans. Hepatology 53:964–973
Yoshinaga K (1987) Uterine receptivity for blastocyst implantation. Ann NY Acad Sci 541:424–431
Acknowledgments
The authors acknowledge Dr Laura A. Lindsay and Mr Samson Dowland for assistance with manuscript editing, Dr Louise Cole (Core Facilities Manager, Bosch Institute Advanced Microscopy Facility, The University of Sydney) for her assistance with the microscopy imaging (Zeiss Deconvolution and Zeiss LSM 510 Metaconfocal microscopes) and Dr Donna Lai (Molecular Biology Officer, Bosch Institute Molecular Biology Facility, The University of Sydney) for her support with western blotting.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Poon, C.E., Madawala, R.J., Day, M.L. et al. EpCAM is decreased but is still present in uterine epithelial cells during early pregnancy in the rat: potential mechanism for maintenance of mucosal integrity during implantation. Cell Tissue Res 359, 655–664 (2015). https://doi.org/10.1007/s00441-014-2017-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-014-2017-3