Abstract
Stripe assays are frequently used for studying binary growth decisions of cells and axons towards surface-bound molecules in vitro. In particular in the fields of neurodevelopment and axon guidance, stripe assays have become a routine tool. Several variants of the stripe assay have been developed since its introduction by Bonhoeffer and colleagues in 1987 (Development 101:685–696, 1987). In all variants, however, the principle is the generation of a structured binary growth substrate, consisting of two sets of cues, arranged in alternating stripes. There are two major classes of stripe assays, mainly distinguished by the source material used for stripe pattern manufacturing: membrane stripe assays, where the stripe patterns are generated with membrane fractions isolated from tissue or cells, and stripe assays with purified proteins, also called modified stripe assays. In this chapter we describe in detail the classical membrane stripe assay, the commonly used modified stripe assay employing purified proteins, and a novel stripe assay for high-affinity interacting proteins, like receptor/ligand pairs.
Key words
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Walter J, Kern-Veits B, Huf J et al (1987) Recognition of position-specific properties of tectal cell membranes by retinal axons in vitro. Development 101:685–696
Walter J, Henke-Fahle S, Bonhoeffer F (1987) Avoidance of posterior tectal membranes by temporal retinal axons. Development 101:909–913
Bonhoeffer F, Huf J (1982) In vitro experiments on axon guidance demonstrating an anterior-posterior gradient on the tectum. EMBO J 1:427–431
Drescher U, Kremoser C, Handwerker C et al (1995) In vitro guidance of retinal ganglion cell axons by RAGS, a 25 kDa tectal protein related to ligands for Eph receptor tyrosine kinases. Cell 82:359–370
Knöll B, Drescher U (2002) Ephrin-As as receptors in topographic projections. Trends Neurosci 25:145–149
McLaughlin T, O’Leary DD (2005) Molecular gradients and development of retinotopic maps. Annu Rev Neurosci 28:327–355
Suetterlin P, Marler KM, Drescher U (2012) Axonal ephrinA/EphA interactions and the emergence of order in topographic projections. Semin Cell Dev Biol 23:1–6
Vielmetter J, Stolze B, Bonhoeffer F et al (1990) In vitro assay to test differential substrate affinities of growing axons and migratory cells. Exp Brain Res 81:283–287
Knöll B, Weinl C, Nordheim A et al (2007) Stripe assay to examine axonal guidance and cell migration. Nat Protoc 2:1216–1224
Rashid T, Upton AL, Blentic A et al (2005) Opposing gradients of ephrin-As and EphA7 in the superior colliculus are essential for topographic mapping in the mammalian visual system. Neuron 47:57–69
Egea J, Klein R (2007) Bidirectional Eph-ephrin signaling during axon guidance. Trends Cell Biol 7:230–238
Feldheim DA, O’Leary DD (2010) Visual map development: bidirectional signaling, bifunctional guidance molecules, and competition. Cold Spring Harb Perspect Biol 2(11):a001768
Gebhardt C, Bastmeyer M, Weth F (2012) Balancing of ephrin/Eph forward and reverse signaling as the driving force of adaptive topographic mapping. Development 139:335–345
von Philipsborn AC, Lang S, Löschinger J et al (2006) Growth cone navigation in substrate-bound ephrin gradients. Development 133:2487–2495
Wizenmann A, Thanos S, von Boxberg Y et al (1993) Differential reaction of crossing and non-crossing rat retinal axons on cell membrane preparations from the chiasm midline: an in vitro study. Development 117:725–735
Hübener M, Götz M, Klostermann S et al (1995) Guidance of thalamocortical axons by growth-promoting molecules in developing rat cerebral cortex. Eur J Neurosci 7:1963–1972
Stein E, Savaskan NE, Ninnemann O et al (1999) A role for the Eph ligand ephrin-A3 in entorhino-hippocampal axon targeting. J Neurosci 19:8885–8893
Monschau B, Kremoser C, Ohta K et al (1997) Shared and distinct functions of RAGS and ELF-1 in guiding retinal axons. EMBO J 16:1258–1267
Simon DK, O’Leary DD (1992) Responses of retinal axons in vivo and in vitro to position-encoding molecules in the embryonic superior colliculus. Neuron 9:977–989
Nakamoto M, Cheng HJ, Friedman GC et al (1996) Topographically specific effects of ELF-1 on retinal axon guidance in vitro and retinal axon mapping in vivo. Cell 86:755–766
Mann F, Ray S, Harris WA et al (2002) Topographic mapping in dorsoventral axis of the Xenopus retinotectal system depends on signaling through ephrin-B ligands. Neuron 35:461–473
Knöll B, Zarbalis K, Wurst W et al (2001) A role for the EphA family in the topographic targeting of vomeronasal axons. Development 128:895–906
Snow DM, Lemmon V, Carrino DA et al (1990) Sulfated proteoglycans in astroglial barriers inhibit neurite outgrowth in vitro. Exp Neurol 109:111–130
Kao TJ, Kania A (2011) Ephrin-mediated cis-attenuation of Eph receptor signaling is essential for spinal motor axon guidance. Neuron 71:76–91
Bonanomi D, Chivatakarn O, Bai G et al (2012) Ret is a multifunctional coreceptor that integrates diffusible- and contact-axon guidance signals. Cell 148:568–582
Forster E, Tielsch A, Saum B et al (2002) Reelin, disabled 1, and beta 1 integrins are required for the formation of the radial glial scaffold in the hippocampus. Proc Natl Acad Sci USA 99:13178–13183
Knöll B, Kretz O, Fiedler C et al (2006) Serum response factor controls neuronal circuit assembly in the hippocampus. Nat Neurosci 9:195–204
Bagnard D, Lohrum M, Uziel D et al (1998) Semaphorins act as attractive and repulsive guidance signals during the development of cortical projections. Development 125:5043–5053
Zimmer G, Schanuel SM, Bürger S et al (2010) Chondroitin sulfate acts in concert with semaphorin 3A to guide tangential migration of cortical interneurons in the ventral telencephalon. Cereb Cortex 20:2411–2422
Wang HU, Anderson DJ (1997) Eph family transmembrane ligands can mediate repulsive guidance of trunk neural crest migration and motor axon outgrowth. Neuron 18:383–396
Cohen RI, Rottkamp DM, Maric D et al (2003) A role for semaphorins and neuropilins in oligodendrocyte guidance. J Neurochem 85:1262–1278
Ciossek T, Monschau B, Kremoser C et al (1998) Eph receptor-ligand interactions are necessary for guidance of retinal ganglion cell axons in vitro. Eur J Neurosci 10:1574–1580
Butler JE (2000) Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays. Methods 22:4–23
Acknowledgments
This work was supported by the German Research Foundation, DFG (grant BA1034/14-3 to M.B. and F.W.). B.K. is supported by the DFG, Schram-Foundation, Gottschalk-Foundation, and Non-Profit Hertie Foundation.
The authors thank Andrea Wizenmann for helpful comments on the manuscript.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Weschenfelder, M., Weth, F., Knöll, B., Bastmeyer, M. (2013). The Stripe Assay: Studying Growth Preference and Axon Guidance on Binary Choice Substrates In Vitro. In: Zhou, R., Mei, L. (eds) Neural Development. Methods in Molecular Biology, vol 1018. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-444-9_22
Download citation
DOI: https://doi.org/10.1007/978-1-62703-444-9_22
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-443-2
Online ISBN: 978-1-62703-444-9
eBook Packages: Springer Protocols