title: Detecting tagged Hedgehog with intracellular and extracellular immunocytochemistry for functional analysis.
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.
authors.forename: Ainhoa
authors.initials: A
authors.lastname: Callejo
authors.forename: Luis
authors.initials: L
authors.lastname: Quijada
authors.forename: Isabel
authors.initials: I
authors.lastname: Guerrero
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004330:Drosophila; D004705:Endocytosis; D005110:Extracellular Space / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D053823:Hedgehog Proteins / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods*; D042541:Intracellular Space / Q000378:metabolism*; D018613:Microscopy, Confocal; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012995:Solubility; D013194:Staining and Labeling; D014921:Wings, Animal / Q000033:anatomy & histology / Q000166:cytology
chemical_list: D053823:Hedgehog Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins