Enhancement of connexin30.3 expression in mouse embryonic stem cell line EB3 in response to cell-cell contacts

To clarify the potential role of gap junction in cell-cell contact response, the expression of connexin30.3 gene (Cx30.3), a specifically expressed isoform in undifferentiated state of mouse embryonic stem (ES) cell line EB3 was investigated under different cell-cell contact conditions. ES cells were cultured by hanging drop culture method to increase cell-cell contact frequency. As control, a single cell culture was conducted. After culture for 12 h, the Cx30.3 expression level in hanging drop culture reached 1.73-fold that of the control (p < 0.001). By contrast, connexin43 gene (Cx43), a ubiquitously expressed gene, showed no difference between both cultures. The experiment of E-cadherin inhibition and β-catenin knockdown suggested the action of E-cadherin upstream of the Cx30.3 regulating pathway. The cell-cell contacts with different cell lines such as HeLa cells and B16/BL6 caused no effect on the Cx30.3 in ES cells. These suggest a potential role of Cx30.3 as a cell-cell contact signal mediator partially regulated by E-cadherin signaling.