Protein insolubility and late-stage morphogenesis in long-term postconfluent cultures of MDCK epithelial cells

Epithelial morphogenesis in vitro has been studied in cultures soon after cell-cell contact at confluency when several actin-associated proteins (fodrin, adherens junction molecules E-cadherin and catenins) localize to specific subcellular domains and become resistant to extraction with non-ionic detergents. Here we demonstrate that early confluency is followed by a long postconfluent period of several weeks during which these proteins and actin itself become progressively enriched in the detergent-resistant fraction of MDCK epithelial cells. Cultures from another tissue (human retinal pigment epithelium) which produces weakly epithelialized monolayers in culture do not exhibit similar late-stage increases in protein insolubility. After confluency some cells in MDCK cultures undergo additional morphogenetic changes giving rise to cord-like structures, and the MDCK adherens junction becomes more stable to disrupting agents. These results indicate that in vitro morphogenesis is not restricted to early confluency in MDCK cells but rather molecular stabilization and dynamic changes in cell shape occur over a protracted postconfluent interval.