Many studies of the main gap junction protein, Connexin43(Cx43), have been explored in porcine oocyte research, but most of studies 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 media and processed for immunocytochemistry and western blotting at 0, 14, 28, and 42h after denuding. In addition, to distinguish whether the tip shaped Cx43 singal near transzonal projections are embedded on oolemma or just exist inside of zona pellucida, we softened zona pellucida by treatment of acidic tyroid for 3 sec to completely divide zona pellucida from plasma membrane. Subsequently we proceeded immunocytochemistry. In result, 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.