title: Assays to Examine Transmembrane Semaphorin Function In Vitro.
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.
authors: Graduate Program in Neuroscience, Department of Cellular and Physiological Sciences, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC, Canada, V6T 1Z3.
authors: Rachel Q
authors: RQ
authors: Liu
authors: Graduate Program in Neuroscience, Department of Cellular and Physiological Sciences, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC, Canada, V6T 1Z3.
authors: Kristen
authors: K
authors: Browne
authors: Graduate Program in Neuroscience, Department of Cellular and Physiological Sciences, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC, Canada, V6T 1Z3. jimo@mail.ubc.ca.
authors: Timothy P
authors: TP
authors: O'Connor
mesh_terms: D000818:Animals; D002642:Chick Embryo; D018920:Coculture Techniques; D003470:Culture Media; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D039961:Semaphorins / Q000502:physiology*; D015398:Signal Transduction
chemical_list: D003470:Culture Media; D039961:Semaphorins
keywords: Cell island; Dissociated neurons; HEK 293 cells; Neuronal explants; Sema5A; Sema5B