Cell Sorting Through E-Cadherin Cleavage

Science Signaling  20 Sep 2011:
Vol. 4, Issue 191, pp. ec263
DOI: 10.1126/scisignal.4191ec263

The sorting and compartmentalization of different cell types are necessary for appropriate tissue development and maintenance. For example, progenitor cells of the intestinal epithelium are localized near the crypt base, and different fates are adopted by the daughter cells as they migrate to different positions. Most cell types of the small intestine migrate upward toward the villus, whereas Paneth cells, which contain secretory granules and antimicrobial peptides, migrate downward to the bottommost position of the crypt base adjacent to intestinal stem cells. Solanas et al. showed that Eph receptors, which are a subgroup of receptor tyrosine kinases that bind to membrane-bound ephrin ligands, interacted with E-cadherin and the membrane-bound metalloproteinase ADAM10 to modulate cell-cell affinity at the boundaries between distinct cell populations. Imaging of canine kidney MDCK cell cocultures composed of cells coexpressing EphB3 and green fluorescent protein (GFP)–tagged E-cadherin and cells expressing Cherry-tagged E-cadherin revealed an intermingling of the two cell populations. In contrast, culturing of MDCK cells coexpressing EphB3 and E-cadherin–GFP with cells coexpressing ephrin-B1 and E-cadherin–Cherry induced the formation of large homogeneous clusters of red or green cells. Endogenous EphB2, E-cadherin, and ADAM10 colocalized throughout the basolateral membrane of MDCK cells. Western blot analysis of supernatants from Co115 colorectal cancer cell cocultures in which cells were expressing either EphB3 or ephrin-B1 revealed ADAM10-mediated cleavage of the extracellular domain of E-cadherin. Moreover, Co115 cells coexpressing EphB3 and either a short hairpin RNA targeting Adam10 or an ADAM10 mutant lacking the metalloproteinase domain (ADAM10ΔMP) did not form large fluorescent protein–labeled homogeneous cell clusters when cocultured with cells expressing ephrin-B1, but rather the cells expressing EphB3 or ephrin-B1 were intermingled. Immunohistochemistry of the intestinal epithelium from transgenic mice expressing ADAM10ΔMP in Paneth cells revealed a scattering of Paneth cells throughout the transient amplifying (TA) compartment and villus, as well as a disorganization of Paneth cells within the crypt base. Thus, the distinct boundary between Paneth cells and the TA compartment that is normally observed in wild-type mice was not formed in the ADAM10ΔMP transgenic mice. These data suggest that localized activation of ADAM10 through EphB–ephrin-B1 interactions on adjacent epithelial cells and subsequent ADAM10-mediated cleavage of E-cadherin in EphB-positive cells modulates cell affinity—and thus cell sorting and compartmentalization.

G. Solanas, C. Cortina, M. Sevillano, E. Batlle, Cleavage of E-cadherin by ADAM10 mediates epithelial cell sorting downstream of EphB signalling. Nat. Cell Biol. 13, 1100–1107 (2011). [PubMed]