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First published online June 8, 2006
doi: 10.1242/10.1242/dev.02412
1 Max-Planck-Institut für Entwicklungsbiologie, Spemannstrasse 35, 72076
Tuebingen, Germany.
2 Universität Karlsruhe (TH), Zoologisches Institut I, Zell- und
Neurobiologie, Haid-und-Neu-Strasse 9, 76131 Karlsruhe, Germany.
3 Institut für Mikro- und Nanotechnologie, Interstaatliche Hochschule
für Technik Buchs NTB, Werdenbergstrasse 4, CH-9471 Buchs,
Switzerland.
4 Labor für Mikro- und Nanotechnologie, Paul Scherrer Institut, CH-5232
Villigen-PSI, Switzerland.
* Authors for correspondence (e-mail: friedich.bonhoeffer{at}tuebingen.mpg.de; bastmeyer{at}bio.uka.de)
Accepted 19 April 2006
Graded distributions of ephrin ligands are involved in the formation of topographic maps. However, it is still poorly understood how growth cones read gradients of membrane-bound guidance molecules. We used microcontact printing to produce discontinuous gradients of substrate-bound ephrinA5. These consist of submicron-sized protein-covered spots, which vary with respect to their sizes and spacings. Growth cones of chick temporal retinal axons are able to integrate these discontinuous ephrin distributions and stop at a distinct zone in the gradient while still undergoing filopodial activity. The position of this stop zone depends on both the steepness of the gradient and on the amount of substrate-bound ephrin per unit surface area. Quantitative analysis of axon outgrowth shows that the stop reaction is controlled by a combination of the local ephrin concentration and the total amount of encountered ephrin, but cannot be attributed to one of these parameters alone.
Key words: Axon guidance, Microcontact printing, Retinotectal projection, Chick
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