Abstract
Eph receptor–ephrin signals are important for controlling repulsive and attractive cell movements during tissue patterning in embryonic development. However, the dynamic cellular responses to these signals at cell–cell contact sites are poorly understood. To examine these events we have used cell microinjection to express EphB4 and ephrinB2 in adjacent Swiss 3T3 fibroblasts and have studied the interaction of the injected cells using time-lapse microscopy. We show that Eph receptors are locally activated wherever neighbouring cells make contact. This triggers dynamic, Rac-regulated membrane ruffles at the Eph–ephrin contact sites. Subsequently, the receptor and ligand cells retract from one another, concomitantly with the endocytosis of the activated Eph receptors and their bound, full-length ephrinB ligands. Both the internalization of the receptor–ligand complexes and the subsequent cell retraction events are dependent on actin polymerization, which in turn is dependent on Rac signalling within the receptor-expressing cells. Similar events occur in primary human endothelial cells. Our findings suggest a novel mechanism for cell repulsion, in which the contact between Eph-expressing and ephrin-expressing cells is destabilized by the localized phagocytosis of the ligand-expressing cell plasma membrane by the receptor-expressing cell.
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Acknowledgements
We thank R. Klein for sharing unpublished results. We also thank R. Adams, D. Wilkinson, M. Henkemeyer, C. Cowan, D. Holowka, L. Machesky, M. Furthmayr and J. M. Edwardson for cDNAs and expression constructs; J. Pendjiky, D. Ciantar and M. Shipman for help with image acquisition and processing; P. Martin and M. Raff for valuable comments on the manuscript, and members of the Nobes laboratory for advice and helpful discussions. This work was inspired by the late Nigel Holder and was supported by the Medical Research Council. C.D.N. is an MRC Senior Research Fellow.
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Marston, D., Dickinson, S. & Nobes, C. Rac-dependent trans-endocytosis of ephrinBs regulates Eph–ephrin contact repulsion. Nat Cell Biol 5, 879–888 (2003). https://doi.org/10.1038/ncb1044
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DOI: https://doi.org/10.1038/ncb1044
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