Heparin-Binding Epidermal Growth Factor-Like Growth Factor Enhances Aquaporin 3 Expression and Function During Mouse Embryo Implantation

Aquaporin 3 (AQP3) is highly expressed in peri-implantation blastocyst trophoblastic cells, indicating its role in cytotrophoblast invasion during embryo implantation. However, the mechanism underlying the regulation of AQP3 expression during embryo implantation remains unclear. In this study, an in vitro co-culture system of blastocysts on a monolayer of uterine endometrial cells was used to mimic in vivo process of embryo attachment and invasion to uterine endometrium and treated with different concentrations of heparin-binding epidermal growth factor-like growth factor (HB-EGF). The results showed that HB-EGF enhanced AQP3 expression in blastocysts in a dose-dependent manner and promoted the attachment and outgrowth of blastocysts on the monolayer of uterine endometrial cells. When the AQP3 activity was inhibited by copper sulfate, both the attachment and outgrowth of blastocysts were inhibited. Furthermore, HB-EGF induced the phosphorylation of EGF receptor (EGFR) and extracellular signal-regulated kinase (ERK). PD153035 (EGFR inhibitor) and U0126 (ERK inhibitor) inhibited AQP3 expression and also the attachment and outgrowth of blastocysts. Collectively, our findings provide the first evidence that HB-EGF stimulates EGFR/ERK signaling to promote AQP3 expression in trophoblastic cells, and AQP3 plays a vital role in HB-EGF-induced embryo implantation. Similar content being viewed by others

Paria BC, Ma WG, Tan J, et al. Cellular and molecular responses of the uterus to embryo implantation can be elicited by locally applied growth factors. Proc Natl Acad Sci U.S.A. 2001;98(3):1047–1052. Bergh PA, Navot D. The impact of embryonic development and endometrial maturity on the timing of implantation. Fertil Steril. 1992;58(3):537–542. Fang L Q, Zhang H, Ding X Y, et al. Mouse trophoblastic cells exhibit a dominant invasiveness phenotype over cancer cells. Cancer Lett. 2010;299(2):111–118. Chen XM, O’Hara SP, Huang BQ, et al. Localized glucose and water influx facilitates Cryptosporidium parvum cellular invasion by means of modulation of host-cell membrane protrusion. Proc Natl Acad Sci U.S.A. 2005;102(18):6338–6343. Condeelis J. Life at the leading edge: the formation of cell protrusions. Annu Rev Cell Biol. 1993;9:411–444. Lauffenburger DA, Horwitz AF. Cell migration: a physically integrated molecular process. Cell. 1996;84(3):359–369. Papadopoulos MC, Saadoun S, Verkman AS. Aquaporins and cell migration. Pflügers Arch. 2008;456(4):693–700. Agre P. The aquaporin water channels. Proc Am Thorac Soc. 2006;3(1):5–13. Borgnia M, Nielsen S, Engel A, Agre P. Cellular and molecular biology of the aquaporin water channels. Annu Rev Biochem. 1999;68:425–458. Agre P, King LS, Yasui M, et al. Aquaporin water channels-from atomic structure to clinical medicine. J Physiol. 2002;542(Pt 1): 3–16. Benga G. Water channel proteins (later called aquaporins) and relatives: past, present, and future. IUBMB Life. 2009;61(2):112–133. Hara-Chikuma M, Verkman AS. Aquaporin-3 facilitates epidermal cell migration and proliferation during wound healing. J Mol Med (Berl). 2008;86(2):221–231. Cao C, Sun Y, Healey S, et al. EGFR-mediated expression of aquaporin-3 is involved in human skin fibroblast migration. Biochem J. 2006;400(2):225–234. Huang Y, Zhu Z, Sun M, et al. Critical role of aquaporin-3 in the human epidermal growth factor-induced migration and proliferation in the human gastric adenocarcinoma cells. Cancer Biol Ther 2010;9(12):1000–1007. Nong Y, Liu F, Chen Y, Wang F. The expression and distribution of aquaporin 3 in mouse embryos before and after vitrification. J Assist Reprod Genet. 2013;30(4):601–606. Yoo HJ, Barlow DH, Mardon HJ. Temporal and spatial regulation of expression of heparin-binding epidermal growth factor-like growth factor in the human endometrium: a possible role in blastocyst implantation. Dev Genet. 1997;21(1):102–108. Leach RE, Khalifa R, Ramirez ND, et al. Multiple roles for heparin-binding epidermal growth factor-like growth factor are suggested by its cell-specific expression during the human endometrial cycle and early placentation. J Clin Endocrinol Metab. 1999;84(9):3355–3363. Grant KS, Wira CR. Effect of mouse uterine stromal cells on epithelial cell transepithelial resistance (TER) and TNFα and TGFβ release in culture. Biol Reprod. 2003;69(3):1091–1098. Dai B, Cao Y, Liu W, et al. Dual roles of progesterone in embryo implantation in mouse. Endocrine. 2003;21(2):123–132. Das SK, Wang XN, Paria BC, et al. Heparin-binding EGF-like growth factor gene is induced in the mouse uterus temporally by the blastocyst solely at the site of its apposition: a possible ligand for interaction with blastocyst EGF-receptor in implantation. Development. 1994;120(5):1071–1083. Richard C, Gao JU, Brown N, et al. Aquaporin water channel genes are differentially expressed and regulated by ovarian steroids during the periimplantation period in the mouse. Endocrinology. 2003;144(4):1533–1541. Lindsay LA, Murphy CR. Redistribution of aquaporins in uterine epithelial cells at the time of implantation in the rat. Acta Histochem. 2004;106(4):299–307. Lobel BL, Levy E, Shelesnyak MC. Studies on the mechanism of nidation. XXXIV. Dynamics of cellular interactions during progestation and implantation in the rat. 3. Implantation. Acta Endocrinol (Copenh). 1967;56(Suppl 123):5. Okada Y, Asahina T, Kobayashi T, Goto J, Terao T. Studies on the mechanism of edematous changes at the endometrial stroma for implantation. Semin Thromb Hemost. 2000;27(2):67–77. Offenberg H, Barcroft LC, Caveney A, Viuff D, Thomsen PD,, Watson AJ. mRNAs encoding aquaporins are present during murine preimplantation development. Mol Reprod Dev. 2000;57(4):323–330. Barcroft LC, Offenberg H, Thomsen P, Watson AJ. Aquaporin proteins in murine trophectoderm mediate transepithelial water movements during cavitation. Dev Biol. 2003;256(2):342–354. Nong Y, Liu F, Chen Y, et al. The expression and distribution of aquaporin 3 in mouse embryos before and after vitrification. J Assist Reprod Genet. 2013;30(4):601–606. Lim HJ, Dey SK. HB-EGF: a unique mediator of embryo-uterine interactions during implantation. Exp Cell Res. 2009;315(4):619–626. Higashiyama S, Abraham JA, Miller J, Fiddes JC, Klagsbrun M. A heparin-binding growth factor secreted by macrophage-like cells that is related to EGF. Science. 1991;251(4996):936–939. Higashiyama S, Lau K, Besner GE, Abraham JA, Klagsbrun M. Structure of heparin-binding EGF-like growth factor. Multiple forms, primary structure, and glycosylation of the mature protein. J Biol Chem. 1992;267(9):6205–6212. Elenius K, Paul S, Allison G, Sun J, Klagsbrun M. Activation of HER4 by heparin binding EGF-like growth factor stimulates chemotaxis but not proliferation. EMBO J. 1997;16(6):1268–1278. Paria BC, Elenius K, Klagsbrun M, Dey SK. Heparin-binding EGF-like growth factor interacts with mouse blastocysts independently of ErbB1: a possible role for heparan sulfate proteoglycans and ErbB4 in blastocyst implantation. Development. 1999;126(9):1997–2005. Zelenina M, Tritto S, Bondar AA, Zelenin S, Aperia A. Copper inhibits the water and glycerol permeability of aquaporin-3. J Biol Chem. 2004;279(50):51939–51943. Zelenina M, Bondar AA, Zelenin S, Aperia A. Nickel and extracellular acidification inhibit the water permeability of human aquaporin-3 in lung epithelial cells. J Biol Chem. 2003;278(32):30037–30043. Tarara R, Enders AC, Hendrickx AG, et al. Early implantation and embryonic development of the baboon: stages 5, 6 and 7. Anat Embryol (Berl). 1987;176(3):267–275. Cao XC, Zhang WR, Cao WF, et al. Aquaporin3 is required for FGF-2-induced migration of human breast cancers. PLoS One. 2013;8(2):e56735. Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Fang, CX., Nong, YQ., Liu, FH. et al. Heparin-Binding Epidermal Growth Factor-Like Growth Factor Enhances Aquaporin 3 Expression and Function During Mouse Embryo Implantation. Reprod. Sci. 24, 463–470 (2017). https://doi.org/10.1177/1933719116657893 Published: Issue date: DOI: https://doi.org/10.1177/1933719116657893