Abstract
The Hedgehog pathway has been shown to be an important developmental signaling pathway in many organisms (Ingham and McMahon. Genes Dev 15:3059–3087, 2001). Recently that work has been extended to developing echinoderm embryos (Walton et al. Dev Biol 331(1):26–37, 2009). Here we describe several methods to perturb the Hedgehog signaling pathway in the sea urchin. These include microinjection of Morpholinos and mRNA constructs as well as treatments with small molecule inhibitors. Finally we provide simple methods for assaying Hedgehog phenotypes in the sea urchin embryo.
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
Ingham P, McMahon A (2001) Hedgehog signaling in animal development: paradigms and principles. Genes Dev 15:3059–3087
Walton KD, Warner J, Hertzler PH et al (2009) Hedgehog signaling patterns mesoderm in the sea urchin. Dev Biol 331(1):26–37
McMahon AP, Ingham PW, Tabin CJ (2003) Developmental roles and clinical significance of hedgehog signaling. Curr Top Dev Biol 53:1–114
Hirokawa N, Tanaka Y, Okada Y et al (2006) Nodal flow and the generation of left-right asymmetry. Cell 125(1):33–45
Hooper JE, Scott MP (2005) Communicating with Hedgehogs. Nat Rev Mol Cell Biol 4:306–317
Wilson CW, Chuang PT (2010) Mechanism and evolution of cytosolic Hedgehog signal transduction. Development 137(13):2079–2094
Walton KD, Croce JC, Glenn TD et al (2006) Genomics and expression profiles of the Hedgehog and Notch signaling pathways in sea urchin development. Dev Biol 300(1): 153–164
Taipale J, Chen JK, Cooper MK et al (2000) Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature 406:1005–1009
Chen JK, Taipale J, Cooper MK et al (2002) Inhibition of Hedgehog signaling by direct binding of cyclopamine to Smoothened. Genes Dev 16:2743–2748
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Foltz KR, Adams NL, Runft LL (2004) Echinoderm eggs and embryos: procurement and culture. In: Ettensohn CE, Wessel GM, Wray G (eds) Methods in cell biology vol 74. Development of sea urchins, Ascidians, and other invertebrate deuterostomes: experimental approaches. Academic, San Diego, CA
Cheers SM, Ettensohn CA (2004) Rapid injection of fertilized eggs. In: Ettensohn CE, Wessel GM, Wray G (eds) Methods in cell biology vol 74. Development of sea urchins, Ascidians, and other invertebrate deuterostomes: experimental approaches. Academic, San Diego, CA
Mas C, Altaba AR (2010) Small molecule modulation of HH-GLI signaling: current leads, trials and tribulations. Biochem Pharm 80(5):712–723
Small J, Rottner K, Hahne P et al (1999) Visualizing the cytoskeleton. Microsc Res Tech 47(1):3–17
Acknowledgements
Support provided by NIH HD RO1-14483, HD PO1-37105, and GM-PO50-81883.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Warner, J.F., McClay, D.R. (2014). Perturbations to the Hedgehog Pathway in Sea Urchin Embryos. In: Carroll, D., Stricker, S. (eds) Developmental Biology of the Sea Urchin and Other Marine Invertebrates. Methods in Molecular Biology, vol 1128. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-974-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-62703-974-1_14
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-973-4
Online ISBN: 978-1-62703-974-1
eBook Packages: Springer Protocols