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Semaphorin Signaling pp 247–263Cite as

Assays to Examine Transmembrane Semaphorin Function In Vitro

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1493))

Abstract

The semaphorins are a large family of secreted and membrane associated proteins that play numerous key roles in the development and function of the nervous system and other tissues. They have been primarily associated with their function as guidance cues in the developing nervous system. In general, semaphorins have been shown to function as inhibitory guidance cues; however there are also numerous examples where they can function as attractive or permissive cues. Thus it is important to employ a variety of assays to test for semaphorin function. While numerous assays have been established for secreted semaphorins, testing the function of transmembrane semaphorins has been challenging. In this chapter we outline two assays that we have used extensively to test their function. In one assay we examine the effect of a constant source of a transmembrane semaphorin on neurite outgrowth and in a second assay we examine whether neurons will actively avoid growing across islands of cells expressing a transmembrane semaphorin. We have found both assays to be relatively easy to perform and useful to test semaphorin function and signaling.

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Acknowledgement

This work was funded by a Natural Sciences and Engineering Research Council discovery grant to T.P.O.C. (NSERC 171387-09).

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Authors and Affiliations

  1. Graduate Program in Neuroscience, Department of Cellular and Physiological Sciences, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC, Canada, V6T 1Z3

    Rachel Q. Liu, Kristen Browne & Timothy P. O’Connor

Authors
  1. Rachel Q. Liu
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  2. Kristen Browne
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  3. Timothy P. O’Connor
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Corresponding author

Correspondence to Timothy P. O’Connor .

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Editors and Affiliations

  1. Departments of Neuroscience and Pharmacology, The University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Jonathan R. Terman

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Liu, R.Q., Browne, K., O’Connor, T.P. (2017). Assays to Examine Transmembrane Semaphorin Function In Vitro. In: Terman, J. (eds) Semaphorin Signaling. Methods in Molecular Biology, vol 1493. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6448-2_18

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  • DOI: https://doi.org/10.1007/978-1-4939-6448-2_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6446-8

  • Online ISBN: 978-1-4939-6448-2

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