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Detecting Tagged Hedgehog with Intracellular and Extracellular Immunocytochemistry for Functional Analysis

  • Luis Quijada
  • Isabel Guerrero
Part of the Methods Inmolecular Biology™ book series (MIMB, volume 397)

Abstract

In this chapter, we explain different strategies to analyze the extracellular Hedgehog (Hh) morphogen distribution and Hh intracellular trafficking by immunohistochemistry techniques. For this purpose, it has been very useful to have a transgenic fly line that expresses a Hh-green fluorescent protein (GFP) fusion protein. These flies can be used to study the way Hh spreads through the anterior compartment where it signals, and analyze in detail how Hh is internalized by its receptor Patched. In addition, this Hh-GFP fusion made without lipid modifications (cholesterol or palmitic acid) can be used to investigate the function of these lipids on Hh in terms of spreading, internalization, and signaling abilities.

Key Words

Hedgehog protein (Hh) green fluorescent protein (GFP) patched (Ptc) wing imaginal disc cells Gal 4 driver UAS vector extracellular labeling 

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Copyright information

© Humana Press Inc., Totowa, NJ 1 2007

Authors and Affiliations

  • Luis Quijada
    • 1
  • Isabel Guerrero
    • 1
  1. 1.Centro de Biología Molecular “Severo Ochoa” (C.S.I.C.-UAM)Universidad Autónoma de Madrid, CantoblancoMadridSpain

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