Abstract
A combination of both in vivo and in vitro techniques is invaluable for studying semaphorin signaling in the avian central nervous system. Here we describe how both types of approaches can be used to compliment each other in order to unravel the role that semaphorins play during embryonic development and elucidate the functional consequences of semaphorin knockdown using RNA interference vectors. We describe and discuss specifically the use of in ovo electroporation and primary oculomotor neuron culture to identify the role of semaphorins in oculomotor neuron migration and assess functional consequences of semaphorin disruption in this system.
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 subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Yazdani U, Terman JR (2006) The Semaphorins. Genome Biol 7(3):211
Fiore R, Puschel AW (2003) The function of semaphorins during nervous system development. Front Biosci 8:484–499
Koncina E et al (2013) Role of Semaphorins during axon growth and guidance. Madame Curie bioscience database [Internet]
Polleux F, Morrow T, Ghosh A (2000) Semaphorin 3A is a chemoattractant for cortical apical dendrites. Nature 404:567–573
Hamburger V, Hamilton H (1992) A series of normal stages in the development of the chick embryo. Dev Dyn 195:231–272
Pekarik V et al (2003) Screening for gene function in chicken embryo using RNAi and electroporation. Nat Biotechnol 1:93–96
Stoekli E (2006) Injection of dsRNA and electroporation in avian embryo. CSH Protoc 1:(4)
Das RM et al (2006) A robust system for RNA interference in the chicken using a modified microRNA operon. Dev Biol 294(2):554–563
Muramatsu T et al (1997) Comparison of three non-viral transfection methods for foreign gene expression in early chicken embryos in ovo. Biochem Biophys Res Commun 230:376–380
Osborne NJ et al (2005) Semaphorin/neuropilin signaling influences the positioning of migratory neural crest cells within the hindbrain region of the chick. Dev Dyn 232(4):939–949
Bron R et al (2004) Functional knockdown of neuropilin-1 in the developing chick nervous system by siRNA hairpins phenocopies genetic ablation in the mouse. Dev Dyn 230:299–308
Takahashi T, Nakamura F, Strittmatter SM (1997) Neuronal and non-neuronal collapsin-1 binding sites in developing chick are distinct from other semaphorin binding sites. J Neurosci 17(23):9183–9193
Zhuang B, Su YS, Sockanathan S (2010) FARP1 promotes the dendritic growth of spinal motor neuron subtypes through transmembrane Semaphorin6A and PlexinA4 signalling. Neuron 61(3):359–372
Ferrario JE et al (2012) Axon guidance in the developing ocular motor system and Duane retraction syndrome depends on Semaphorin signaling via alpha2-chimaerin. Proc Natl Acad Sci U S A 109(36):14669–14674
Miyake N et al (2009) Human CHN1 mutations hyperactivate α2-chimaerin and cause Duane’s retraction syndrome. Science 321(5890):839–843
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Montague, K., Guthrie, S., Poparic, I. (2017). In Vivo and In Vitro Knockdown Approaches in the Avian Embryo as a Means to Study Semaphorin Signaling. 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_29
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6448-2_29
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6446-8
Online ISBN: 978-1-4939-6448-2
eBook Packages: Springer Protocols