E-Cadherin and tight junctions between epithelial cells of different animal species

Abstract

The assembly and permanent sealing of tight junctions (TJs) depend crucially on cell-cell contacts containing E-cadherin. This poses a puzzling problem because, while TJs can be established between epithelial cells from different tissues and even different animal species ("heterotypic TJs"; Gonzalez-Mariscal et al. 1989, J Membr Biol 107:43), the cell-cell binding mediated by E-cadherin is a highly specific one (Takeichi 1995, Curr Opin Cell Biol 7:619). Yet the demonstration that TJs can be established at heterotypic borders is open to two distinct challenges. First, it is based on transepithelial electrical resistance (TER) and restriction to ruthenium red permeation only, which today are known to be just two of the many characteristics of TJs; and second some attributes of the TJs (e.g. the presence of specific molecules) have been found even in cells that do not establish these structures. This raised the question of whether heterotypic TJs were not true or full TJs. In the present work we demonstrate that heterotypic TJs in mixed monolayers of MDCK cells with a different cell type (LLC-PK1) are true TJs through several criteria, such as TER, the ability to stop the membrane diffusion of fluorescent sphingomyelin from the apical to the lateral domain, the presence of ZO-1, ZO-2, occludin, claudin-1 and claudin-2. We then turn to the presence of E-cadherin at heterotypic borders, and observe that it cannot be detected by the highly specific DECMA-1 antibody, in spite of the fact that this antibody does reveal the presence of E-cadherin at homotypic contacts of the same cell. Yet, ECCD-2, an antibody against another domain of E-cadherin, reveals that this molecule may be present at both types of borders. Thus, E-cadherin is present at heterotypic borders, yet it seems to be in a conformation unable to bind DECMA-1. Our results suggest: (1) that heterotypic borders can establish fully developed TJs; (2) that the sealing of these heterotypic TJs depends on E-cadherin; (3) but that this dependence is mediated through a cascade of chemical reactions involving two different G-proteins, PLC, PKC and calmodulin, which we have characterized elsewhere (Balda et al. 1991, J Membr Biol 122:193); and (4) hence molecules of E-cadherin that trigger junction formation can act from a distant homotypic contact.

Rotavirus-Induced Early Activation of the RhoA/ROCK/MLC Signaling Pathway Mediates the Disruption of Tight Junctions in Polarized MDCK Cells

“…TJs can also restrict the exchange of membrane molecules, proteins and lipids, between the apical and basolateral domains of the cell membrane, which is termed fence function 6 . The fence function of the TJ can be evaluated in monolayers cultured on transwell membranes by adding a fluorescent lipid (BodipyFL-C12-sphingomyelin-BSA complex) onto the apical membrane and observing whether the fluorescent label reaches the basolateral membrane 7 , 46 , 47 . Therefore, mock-pretreated or pretreated MDCK monolayers grown on transwell filters were either mock-infected or infected with RVA strain DS-1 or NDCV for 1 h. The apical compartments were then incubated with BodipyFL-C12-sphingomyelin/BSA complex, and the cell membranes were immediately analyzed by confocal microscopy.…”

“…Labelling of cells with fluorescent lipids was performed as previously described 7 , 46 , 47 . Briefly, polarized MDCK cells grown on polyester membrane transwell inserts were treated with or without chemicals as well as infected with or without RVA strain DS-1 or NCDV (MOI = 10).…”

The Polarized Distribution of Na+,K+-ATPase: Role of the Interaction between β Subunits

“…TJs are promiscuous, because those established between cells belonging to different animal species have a degree of sealing that can be predicted from the transepithelial electrical resistance of each cell type and their proportions in the mixed monolayer (González-Mariscal et al , 1989). In contrast, adherens junctions are homotypic and can be seldom, if ever, established between epithelial cells from different animal species (Contreras et al , 2002; Halbleib and Nelson, 2006). We have found previously that the homotypic requirement for cell–cell junctions extends to the distribution of Na + ,K + -ATPase (Contreras et al , 1995).…”

p38 MAPK mediates calcium oxalate crystal-induced tight junction disruption in distal renal tubular epithelial cells

“…Adherens junction has been known as a crucial structure for tight junction formation4849. Furthermore, there is evidence indicating that activation of p38 MAPK can lead to disruption of adherens junction50.…”