Hedgehog Signaling Protocols

In both vertebrates and invertebrates, Hedgehog signaling regulates the development of the tissues and structures of the body. In addition, many cancers and diseases arise as a result of misregulation of the pathway, which makes it an incredibly important system to understand. This volume examines various methods used in the analysis of the Hedgehog signaling pathway, bringing together different lines of analyses into one accessible and comprehensive text. This volume provides a single source for investigators, presenting several different experimental approaches, which are all varied and broad in nature, as well as procedures for a variety of different model systems. Researchers will find this volume to be an invaluble tool as they work towards deeper understanding of the underlying molecular events that the Hedgehog signaling pathway regulates.

Offers various detailed chapters on experimental description, including anecdotal pointers and notes Intended for Molecular Biologists, Biochemists, and Geneticists

Inhalt
Purifying the Hedgehog Protein and its Variants.- Application of Sonic Hedgehog to the Developing Chick Limb.- Manipulation of Hedgehog Signaling in Xenopus by Means of Embryo Microinjection and Application of Chemical Inhibitors.- Isolation of Rat Telencephalic Neural Explants to Assay Shh GABAergic Interneuron Differentiation-Inducing Activity.- Genetic Analysis of the Vertebrate Hedgehog-Signaling Pathway Using Muscle Cell Fate Specification in the Zebrafish Embryo.- Efficient Manipulation of Hedgehog/GLI Signaling Using Retroviral Expression Systems.- Cell Surface Marker and Cell Cycle Analysis, Hedgehog Signaling, and Flow Cytometry.- Detecting Tagged Hedgehog with Intracellular and Extracellular Immunocytochemistry for Functional Analysis.- Confocal Analysis of Hedgehog Morphogenetic Gradient Coupled with Fluorescent In Situ Hybridization of Hedgehog Target Genes.- RNAi in the Hedgehog Signaling Pathway: pFRiPE, a Vector for Temporally and Spatially Controlled RNAi in Drosophila.- Germline Clone Analysis for Maternally Acting Drosophila Hedgehog Components.- Clonal Analysis of Hedgehog Signaling in Drosophila Somatic Tissues.- GAL4/UAS Targeted Gene Expression for Studying Drosophila Hedgehog Signaling.- Biochemical Fractionation of Drosophila Cells.- Using Immunoprecipitation to Study ProteinProtein Interactions in the Hedgehog-Signaling Pathway.- Sequence Analyses to Study the Evolutionary History and Cis-Regulatory Elements of Hedgehog Genes.