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Expression of ZO-1 and occludin at mRNA and protein level during preimplantation development of the pig parthenogenetic diploids

Published online by Cambridge University Press:  22 February 2011

Shangdan Xu ,
Jibak Lee  and
Masashi Miyake
Shangdan Xu
Affiliation:
Department of Bioresource Sciences, Graduate School of Science and Technology, Kobe University, Nada-ku, Kobe 657–8501, Japan.
Jibak Lee
Affiliation:
Chromosome Dynamics Laboratory, RIKEN Advanced Science Institute, Wako, Saitama 351–0198, Japan.
Masashi Miyake*
Affiliation:
Department of Bioresource Sciences, Graduate School of Science and Technology, Kobe University, Nada-ku, Kobe 657–8501, Japan. Organization of Advanced Science and Technology, Kobe University, Nada-ku, Kobe 657–8501, Japan.
*
All correspondence to: Masashi Miyake. Department of Bioresource Sciences, Graduate School of Science and Technology, Kobe University, Nada-ku, Kobe 657–8501, Japan. Tel: +81–78–803–6581. Fax: +81–78–803–5807. e-mail address: miyake@kobe-u.ac.jp
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Summary

Expression of mRNAs and proteins of ZO-1 and occludin was analyzed in pig oocytes and parthenogenetic diploid embryos during preimplantation development using real-time RT-PCR, western blotting and immunocytochemistry. All germinal vesicle (GV) and metaphase (M)II oocytes and preimplantation embryos expressed mRNAs and proteins of ZO-1 and occludin. mRNA levels of both ZO-1 and occludin decreased significantly from GV to MII, but increased at the 2-cell stage followed by temporal decrease during the early and late 4-cell stages. Then, both mRNAs increased after compaction. Relative concentration of zo1α− was highest in 2-cell embryos, while zo1α+ was expressed from the morula stage. Occludin expression greatly increased after the morula stage and was highest in expanded blastocysts. Western blotting analysis showed constant expression of ZO-1α− throughout preimplantation development and limited translation of ZO-1α+ from the blastocysts, and species-specific expression pattern of occludin. Immunocytochemistry analysis revealed homogeneous distribution of ZO-1 and occludin in the cytoplasm with moderately strong fluorescence in the vicinity of the contact region between blastomeres, around the nuclei in the 2-cell to late 4-cell embryos, and clear network localization along the cell-boundary region in embryos after the morula stage. Present results show that major TJ proteins, ZO-1 and occludin are expressed in oocytes and preimplantation embryos, and that ZO-1α+ is transcribed by zygotic gene activation and translated from early blastocysts with prominent increase of occludin at the blastocyst stage.

Keywords

Parthenogenetic embryoPigPreimplantationTight junction
Type
Research Article
Information
Zygote , Volume 20 , Issue 2 , May 2012 , pp. 147 - 158
Copyright
Copyright © Cambridge University Press 2011

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References

Angelow, S., Ahlstrom, R. & Yu, A.S. (2008). Biology of claudins. Am. J. Physiol. Renal Physiol. 295, F867–76.Google Scholar
Barcroft, L.C., Hay-Schmidt, A., Caveney, A., Gilfoyle, E., Overstrom, E.W., Hyttel, P. & Watson, A.J. (1998). Trophectoderm differentiation in the bovine embryo: characterization of a polarized epithelium. J. Reprod. Fertil. 114, 327–39.CrossRefGoogle ScholarPubMed
Bazzoni, G., Martinez-Estrada, O.M., Orsenigo, F., Cordenonsi, M., Citi, S. & Dejana, E. (2000). Interaction of junctional adhesion molecule with the tight junction components ZO-1, cingulin, occludin. J. Biol. Chem. 275, 20520–6.CrossRefGoogle ScholarPubMed
Biggers, J.D., Bell, J.E. & Benos, D.J. (1988). Mammalian blastocyst: transport functions in a developing epithelium. Am. J. Physiol. 255, C419–32.CrossRefGoogle Scholar
Collins, J.E. & Fleming, T.P. (1995). Epithelial differentiation in the mouse preimplantation embryo: making adhesive cell contacts for the first time. Trends Biochem. Sci. 20, 307–12.CrossRefGoogle ScholarPubMed
Cordenonsi, M., Mazzon, E., De Rigo, L., Baraldo, S., Meggio, F. & Citi, S. (1997). Occludin dephosphorylation in early development of Xenopus laevis. J. Cell Sci. 110, 3131–9.CrossRefGoogle Scholar
Eckert, J.J. & Fleming, T.P. (2008). Tight junction biogenesis during early development. Biochim. Biophys. Acta 1778, 717–28.Google Scholar
Eckert, J.J., Houghton, F.D., Hawkhead, J.A., Balen, A.H., Leese, H.J., Picton, H.M., Cameron, I.T. & Fleming, T.P. (2007). Human embryos developing in vitro are susceptible to impaired epithelial junction biogenesis correlating with abnormal metabolic activity. Hum. Reprod. 22, 2214–24.CrossRefGoogle ScholarPubMed
Feldman, G.J., Mullin, J.M. & Ryan, M.P. (2005). Occludin: structure, function and regulation. Adv. Drug Deliv. Rev. 57, 883–917.CrossRefGoogle ScholarPubMed
Fleming, T.P., McConnell, J., Johnson, M.H. & Stevenson, B.R. (1989). Development of tight junctions de novo in the mouse early embryo: control of assembly of the tight junction-specific protein, ZO-1. J. Cell Biol. 108, 1407–18.Google Scholar
Furuse, M., Hirase, T., Itoh, M., Nagafuchi, A., Yonemura, S., Tsukita, S. & Tsukita, S. (1993). Occludin: a novel integral membrane protein localizing at tight junctions. J. Cell Biol. 123, 1777–88.CrossRefGoogle ScholarPubMed
Ghassemifar, M.R., Eckert, J.J., Houghton, F.D., Picton, H.M., Leese, H.J. & Fleming, T.P. (2003). Gene expression regulating epithelial intercellular junction biogenesis during human blastocyst development in vitro. Mol. Hum. Reprod. 9, 245–52.CrossRefGoogle ScholarPubMed
Hirase, T., Staddon, J.M., Saitou, M., Ando-Akatsuka, Y., Itoh, M., Furuse, M., Fujimoto, K., Tsukita, S. & Rubin, L.L. (1997). Occludin as a possible determinant of tight junction permeability in endothelial cells. J. Cell Sci. 110, 1603–13.CrossRefGoogle ScholarPubMed
Holm, P., Booth, P.J. & Callesen, H. (2002). Kinetics of early in vitro development of bovine in vivo- and in vitro-derived zygotes produced and/or cultured in chemically defined or serum-containing media. Reproduction 123, 553–65.Google Scholar
Hwang, K.C., Lee, H.Y., Cui, X.S., Kim, J.H. & Kim, N.H. (2005). Identification of maternal mRNAs in porcine parthenotes at the 2-cell stage: a comparison with the blastocyst stage. Mol. Reprod. Dev. 70, 314–23.Google Scholar
Hyafil, F., Morello, D., Babinet, C. & Jacob, F. (1980). A cell surface glycoprotein involved in the compaction of embryonal carcinoma cells and cleavage stage embryos. Cell 21, 927–34.CrossRefGoogle ScholarPubMed
Itoh, M., Nagafuchi, A., Yonemura, S., Kitani-Yasuda, T., Tsukita, S. & Tsukita, S. (1993). The 220-kD protein colocalizing with cadherins in non-epithelial cells is identical to ZO-1, a tight junction-associated protein in epithelial cells: cDNA cloning and immunoelectron microscopy. J. Cell Biol. 121, 491–502.Google Scholar
Katsuno, T., Umeda, K., Matsui, T., Hata, M., Tamura, A., Itoh, M., Takeuchi, K., Fujimori, T., Nabeshima, Y., Noda, T., Tsukita, S. & Tsukita, S. (2008). Deficiency of zonula occludens-1 causes embryonic lethal phenotype associated with defected yolk sac angiogenesis and apoptosis of embryonic cells. Mol. Biol. Cell 19, 2465–75.Google Scholar
Komiya, S., Shimizu, M., Ikenouchi, J., Yonemura, S., Matsui, T., Fukunaga, Y., Liu, H., Endo, F., Tsukita, S. & Nagafuchi, A. (2005). Apical membrane and junctional complex formation during simple epithelial cell differentiation of F9 cells. Genes Cells 10, 1065–80.Google Scholar
Kuijk, E.W., du Puy, L., van Tol, H.T., Haagsman, H.P., Colenbrander, B. & Roelen, B.A. (2007). Validation of reference genes for quantitative RT-PCR studies in porcine oocytes and preimplantation embryos. BMC Dev. Biol. 7, 58.CrossRefGoogle ScholarPubMed
Kure-bayashi, S., Miyake, M., Katayama, M., Miyano, T. & Kato, S. (1995). Improvement of developmental ability to the blastocyst stage by addition of hyaluronic acid to chemically defined medium in diploid porcine eggs matured in-vitro and subsequently electro-activated. J. Mamm. Ova Res. 12, 119–25.Google Scholar
Kure-bayashi, S., Miyake, M., Katayama, M., Miyano, T. & Kato, S. (1996). Development of porcine blastocysts from in vitro-matured and activated haploid and diploid oocytes. Theriogenology 46, 1027–36.Google Scholar
Kure-bayashi, S., Miyake, M., Okada, K. & Kato, S. (2000). Successful implantation of in vitro-matured, electro-activated oocytes in the pig. Theriogenology 53, 1105–19.CrossRefGoogle ScholarPubMed
Larue, L., Ohsugi, M., Hirchenhain, J. & Kemler, R. (1994). E-cadherin null mutant embryos fail to form a trophectoderm epithelium. Proc. Natl. Acad. Sci. USA 91, 8263–7.CrossRefGoogle Scholar
Magnani, L. & Cabot, R.A. (2008). In vitro and in vivo derived porcine embryos possess similar, but not identical, patterns of Oct4, Nanog, and Sox2 mRNA expression during cleavage development. Mol. Reprod. Dev. 75, 1726–35.CrossRefGoogle Scholar
Martin, T.A. & Jiang, W.G. (2009). Loss of tight junction barrier function and its role in cancer metastasis. Biochim. Biophys. Acta 1788, 872–91.CrossRefGoogle ScholarPubMed
Miller, D.J., Eckert, J.J., Lazzari, G., Duranthon-Richoux, V., Sreenan, J., Morris, D., Galli, C., Renard, J.P. & Fleming, T.P. (2003). Tight junction messenger RNA expression levels in bovine embryos are dependent upon the ability to compact and in vitro culture methods. Biol. Reprod. 68, 1394–402.CrossRefGoogle ScholarPubMed
Moriwaki, K., Tsukita, S. & Furuse, M. (2007). Tight junctions containing claudin 4 and 6 are essential for blastocyst formation in preimplantation mouse embryos. Dev. Biol. 312, 509–22.CrossRefGoogle ScholarPubMed
Muresan, Z., Paul, D.L. & Goodenough, D.A. (2000). Occludin 1B, a variant of the tight junction protein occludin. Mol. Biol. Cell 11, 627–34.CrossRefGoogle ScholarPubMed
Ohsugi, M., Hwang, S., Butz, S., Knowles, B.B., Solter, D. & Kewler, R. (1996). Expression and cell membrane localization of catenins during mouse preimplantation development. Dev. Dynamics 206, 391–402.3.0.CO;2-D>CrossRefGoogle ScholarPubMed
Saitou, M., Furuse, M., Sasaki, H., Schulzke, J.D., Fromm, M., Takano, H., Noda, T. & Tsukita, S. (2000). Complex phenotype of mice lacking occludin, a component of tight junction strands. Mol. Biol. Cell 11, 4131–42.CrossRefGoogle ScholarPubMed
Sheth, B., Fesenko, I., Collins, J.E., Moran, B., Wild, A.E., Anderson, J.M. & Fleming, T.P. (1997). Tight junction assembly during mouse blastocyst formation is regulated by late expression of ZO-1 alpha+ isoform. Development 124, 2027–37.CrossRefGoogle ScholarPubMed
Sheth, B., Fontaine, J.-J., Ponza, E., McCallum, A., Page, A., Citi, S., Louvard, D., Zahraoui, A. & Fleming, T.P. (2000a). Differentiation of the epithelial apical junctional complex during mouse preimplantation development: a role for rab13 in the early maturation of the tight junction. Mech. Dev. 97, 93–104.Google Scholar
Sheth, B., Moran, B., Anderson, J.M. & Fleming, T.P. (2000b). Post-translational control of occludin membrane assembly in mouse trophectoderm: a mechanism to regulate timing of tight junction biogenesis and blastocyst formation. Development 127, 831–40.CrossRefGoogle ScholarPubMed
Sheth, B., Nowak, R.L., Anderson, R., Kwong, W.Y., Papenbrock, T. & Fleming, T.P. (2008). Tight junction protein ZO-2 expression and relative function of ZO-1 and ZO-2 during mouse blastocyst formation. Exp. Cell Res. 314, 3356–68.CrossRefGoogle ScholarPubMed
Stevenson, B.R., Siliciano, J.D., Mooseker, M.S. & Goodenough, D.A. (1986). Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia. J. Cell Biol. 103, 755–66.CrossRefGoogle Scholar
Thuan, N.V., Harayama, H. & Miyake, M. (2002). Characteristics of preimplantational development of porcine parthenogenetic diploids relative to the existence of amino acids in vitro. Biol. Reprod. 67, 1688–98.CrossRefGoogle Scholar
Thuan, N.V., Kure-bayashi, S., Harayama, H., Nagai, T. & Miyake, M. (2003). Stage-specific effects of the osmolarity of a culture medium on the development of parthenogenetic diploids in the pig. Theriogenology 59, 719–34.Google Scholar
Tsukita, S., Furuse, M. & Itoh, M. (2001). Multifunctional strands in tight junctions. Nat. Rev. Mol. Cell. Biol. 2, 285–93.CrossRefGoogle ScholarPubMed
Tsukita, S., Yamazaki, Y., Katsuno, T., Tamura, A. & Tsukita, S. (2008). Tight junction-based epithelial microenvironment and cell proliferation. Oncogene 27, 6930–8.CrossRefGoogle ScholarPubMed
Van Itallie, C.M. & Anderson, J.M. (2006). Claudins and epithelial paracellular transport. Annu. Rev. Physiol. 68, 403–29.Google Scholar
Wang, H., Ding, T., Brown, N., Yamamoto, Y., Prince, L.S., Reese, J. & Paria, B.C. (2008). Zonula occludens-1 (ZO-1) is involved in morula to blastocyst transformation in the mouse. Dev. Biol. 318, 112–25.CrossRefGoogle ScholarPubMed
Watson, A.J., Natale, D.R. & Barcroft, L.C. (2004). Molecular regulation of blastocyst formation. Anim. Reprod. Sci. 82–83, 583–92.CrossRefGoogle ScholarPubMed
Willott, E., Balda, M.S., Heintzelman, M., Jameson, B. & Anderson, J.M. (1992). Localization and differential expression of two isoforms of the tight junction protein ZO-1. Am. J. Physiol. 262, C1119–24.CrossRefGoogle ScholarPubMed
Xu, J., Kausalya, P.J., Phua, D.C., Ali, S.M., Hossain, Z. & Hunziker, W. (2008). Early embryonic lethality of mice lacking ZO-2, but not ZO-3, reveals critical and nonredundant roles for individual zonula occludens proteins in mammalian development. Mol. Cell. Biol. 28, 1669–78.CrossRefGoogle Scholar
Yamanaka, Y., Ralston, A., Stephenson, R.O. & Rossant, J. (2006). Cell and molecular regulation of the mouse blastocyst. Dev. Dyn. 235, 2301–14.Google Scholar
Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I.M. & Iwamura, S. (2002). Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol. Reprod. 66, 112–9.Google Scholar