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Localization and quantitative analysis of Cx43 in porcine oocytes during in vitro maturation

Published online by Cambridge University Press:  21 July 2015

Seunghoon Lee ,
Yuuki Hiradate ,
Yumi Hoshino ,
Yeoung-gyu Ko ,
Kentaro Tanemura  and
Eimei Sato
Seunghoon Lee
Affiliation:
Animal Biotechnology Division, National Institute of Animal Science, RDA, Wanju, 565–851Korea.
Yuuki Hiradate
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan.
Yumi Hoshino
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan.
Yeoung-gyu Ko
Affiliation:
Animal Biotechnology Division, National Institute of Animal Science, RDA, Wanju, 565–851Korea.
Kentaro Tanemura*
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981–8555, Japan. Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan.
Eimei Sato
Affiliation:
National Livestock Breeding, Fukushima, Japan.
*
All correspondence to: Kentaro Tanemura, Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981–8555, Japan. Tel:/Fax: +81 22 717 8686. E-mail: kentaro@m.tohoku.ac.jp
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Summary

Many studies of the main gap junction protein, Cx43, have been conducted in porcine oocyte research, but they have been limited to investigations of cumulus–oocyte complexes (COCs). In this study, we verified Cx43 not in COCs, but in porcine oocytes during maturation, and conducted a quantitative time course analysis. The location and dynamics of Cx43 were examined by immunocytochemistry and western blotting, respectively. COCs were cultured in NCSU23 medium and processed for immunocytochemistry and western blotting at 0, 14, 28, and 42 h after denuding. A Cx43 signal was detected on oolemmas, transzonal projections and the surface of zona pellucidae. Western blotting showed that Cx43 band density increased from 0 to 14 h, and gradually decreased thereafter. Our results clarified that Cx43 is localized in the ooplasmic membrane through zona pellucidae and its level changes over time during culture in porcine oocytes.

Keywords

Connexin43LocalizationPorcine oocyteQuantitative analysis
Type
Research Article
Information
Zygote , Volume 24 , Issue 3 , June 2016 , pp. 364 - 370
Copyright
Copyright © Cambridge University Press 2015 

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References

Abeydeera, L.R., Wang, W.H., Cantley, T.C., Rieke, A., Prather, R.S. & Day, B.N. (1998). Presence of epidermal growth factor during in vitro maturation of pig oocytes and embryo culture can modulate blastocyst development after in vitro fertilization. Mol. Reprod. Dev. 51, 395–401.3.0.CO;2-Y>CrossRefGoogle ScholarPubMed
Anderson, E. & Albertini, D.F. (1976). Gap junctions between the oocyte and companion follicle cells in the mammalian ovary. J. Cell Biol. 71, 680–6.Google Scholar
Anderson, E., Wilkinson, R.F., Lee, G. & Meller, S. (1978). A correlative microscopical analysis of differentiating ovarian follicles of mammals. J. Morphol. 156, 339–66.Google Scholar
Appeltant, R., Somfai, T., Nakai, M., Bodo, S., Maes, D., Kikuchi, K. & Van Soom, A. (2015). Interactions between oocytes and cumulus cells during in vitro maturation of porcine cumulus–oocyte complexes in a chemically defined medium: Effect of denuded oocytes on cumulus expansion and oocyte maturation. Theriogenology 83, 567–76.CrossRefGoogle Scholar
Bejarano, E., Girao, H., Yuste, A., Patel, B., Marques, C., Spray, D.C., Pereira, P. & Cuervo, A.M. (2012). Autophagy modulates dynamics of connexins at the plasma membrane in a ubiquitin-dependent manner. Mol. Biol. Cell 23, 2156–69.Google Scholar
Berthoud, V.M., Minogue, P.J., Osmolak, P., Snabb, J.I. & Beyer, E.C. (2014). Roles and regulation of lens epithelial cell connexins. FEBS Lett. 588, 1297–303.Google Scholar
Chen, H., Zhao, L., Chu, G., Kito, G., Yamauchi, N., Shigeyoshi, Y., Hashimoto, S. & Hattori, M.A. (2013). FSH induces the development of circadian clockwork in rat granulosa cells via a gap junction protein Cx43-dependent pathway. Am. J. Physiol. Endocrinol. Metab. 304, E566–75.CrossRefGoogle Scholar
Fernandes, R., Girao, H. & Pereira, P. (2004). High glucose down-regulates intercellular communication in retinal endothelial cells by enhancing degradation of connexin 43 by a proteasome-dependent mechanism. J. Biol. Chem. 279, 27219–24.Google Scholar
Fong, J.T., Kells, R.M., Gumpert, A.M., Marzillier, J.Y., Davidson, M.W. & Falk, M.M. (2012). Internalized gap junctions are degraded by autophagy. Autophagy, 8, 794–811.CrossRefGoogle ScholarPubMed
Granot, I. & Dekel, N. (2002). The ovarian gap junction protein connexin43: regulation by gonadotropins. Trends Endocrinol. Metab. 13, 310–3.CrossRefGoogle ScholarPubMed
Grosely, R. & Sorgen, P.L. (2013). A history of gap junction structure: hexagonal arrays to atomic resolution. Cell Commun. Adhesion 20, 11–20.CrossRefGoogle ScholarPubMed
Guy, S., Geletu, M., Arulanandam, R. & Raptis, L. (2014). Stat3 and gap junctions in normal and lung cancer cells. Cancers 6, 646–62.CrossRefGoogle ScholarPubMed
Herzog, D.P., Dohle, E., Bischoff, I. & Kirkpatrick, C.J. (2014). Cell communication in a coculture system consisting of outgrowth endothelial cells and primary osteoblasts. BioMed. Res. Int. 2014, 320123.Google Scholar
Kempisty, B., Ziolkowska, A., Piotrowska, H., Antosik, P., Bukowska, D., Zawierucha, P., Jaskowski, J.M., Brussow, K.P., Nowicki, M. & Zabel, M. (2014). Expression and cellular distribution of cyclin-dependent kinase 4 (Cdk4). and connexin 43 (Cx43). in porcine oocytes before and after in vitro maturation. Acta Veterinaria Hungarica 62, 84–95.CrossRefGoogle ScholarPubMed
Kidder, G.M. & Mhawi, A.A. (2002). Gap junctions and ovarian folliculogenesis. Reproduction 123, 613–20.Google Scholar
Leithe, E. & Rivedal, E. (2004). Epidermal growth factor regulates ubiquitination, internalization and proteasome-dependent degradation of connexin 43. J. Cell Sci. 117, 1211–20.CrossRefGoogle Scholar
Li, R. & Albertini, D.F. (2013). The road to maturation: somatic cell interaction and self-organization of the mammalian oocyte. Nature reviews. Mol. Cell. Biol. 14, 141–52.Google Scholar
Louros, N.N., Iconomidou, V.A., Giannelou, P. & Hamodrakas, S.J. (2013). Structural analysis of peptide-analogues of human zona pellucida ZP1 protein with amyloidogenic properties: insights into mammalian zona pellucida formation. PLoS One 8, e73258.Google Scholar
Makita, M. & Miyano, T. (2014). Steroid hormones promote bovine oocyte growth and connection with granulosa cells. Theriogenology 82, 605–12.Google Scholar
Nielsen, M.S., Axelsen, L.N., Sorgen, P.L., Verma, V., Delmar, M. & Holstein-Rathlou, N.H. (2012). Gap junctions. Comp. Physiol. 2, 1981–2035.CrossRefGoogle ScholarPubMed
Oshima, A. (2014). Structure and closure of connexin gap junction channels. FEBS Lett. 588, 1230–7.Google Scholar
Palma, G.A., Arganaraz, M.E., Barrera, A.D., Rodler, D., Mutto, A.A. & Sinowatz, F. (2012). Biol. and biotechnology of follicle development. Sci. World J. 2012, 938138.CrossRefGoogle Scholar
Pandey, A., Gupta, N. & Gupta, S.C. (2009). Improvement of in vitro oocyte maturation with lectin supplementation and expression analysis of Cx43, GDF-9, FGF-4 and fibronectin mRNA transcripts in buffalo (Bubalus bubalis). J. Assist. Reprod. Genet. 26, 365–71.Google Scholar
Pant, D., Reynolds, L.P., Luther, J.S., Borowicz, P.P., Stenbak, T.M., Bilski, J.J., Weigl, R.M., Lopes, F., Petry, K., Johnson, M.L., Redmer, D.A. & Grazul-Bilska, A.T. (2005). Expression of connexin 43 and gap junctional intercellular communication in the cumulus-oocyte complex in sheep. Reproduction 129, 191–200.Google Scholar
Petters, R.M. & Wells, K.D. (1993). Culture of pig embryos. J. Reprod. Fertil. 48 (Suppl.), 61–73.Google Scholar
Quinn, P., Barros, C. & Whittingham, D.G. (1982). Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa. J. Reprod. Fertil. 66, 161–8.Google Scholar
Santiquet, N., Robert, C. & Richard, F.J. (2013). The dynamics of connexin expression, degradation and localisation are regulated by gonadotropins during the early stages of in vitro maturation of swine oocytes. PLoS One 8, e68456.Google Scholar
Shimada, M. & Terada, T. (2001). Phosphatidylinositol 3-kinase in cumulus cells and oocytes is responsible for activation of oocyte mitogen-activated protein kinase during meiotic progression beyond the meiosis I stage in pigs. Biol. Reprod. 64, 1106–14.Google Scholar
Shimada, M., Maeda, T. & Terada, T. (2001). Dynamic changes of connexin-43, gap junctional protein, in outer layers of cumulus cells are regulated by PKC and PI 3-kinase during meiotic resumption in porcine oocytes. Biol. Reprod. 64, 1255–63.Google Scholar
Simon, A.M., Chen, H. & Jackson, C.L. (2006). Cx37 and Cx43 localize to zona pellucida in mouse ovarian follicles. Cell Commun. Adhesion 13, 61–77.Google Scholar
Solan, J.L. & Lampe, P.D. (2009). Connexin43 phosphorylation: structural changes and biological effects. Biochem. J. 419, 261–72.Google Scholar
Verheule, S. & Kaese, S. (2013). Connexin diversity in the heart: insights from transgenic mouse models. Front. Pharmacol. 4, 81.CrossRefGoogle ScholarPubMed
Wang, Q., Chi, M.M., Schedl, T. & Moley, K.H. (2012). An intercellular pathway for glucose transport into mouse oocytes. Am. J. Physiol. Endocrinol. Metab. 302, E1511–8.CrossRefGoogle ScholarPubMed