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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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