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Methods in cell biology. v.94, Primary cilia / edited by Roger D. Sloboda. — Amsterdam : Academic Press, 2009. – (58.1574/M592/v.94)

Contents

    CONTENTS
    
    Contributors
    Preface
    SECTION I Background
    1. From Central to Rudimentary to Primary: The History of an Underappreciated Organelle Whose Time Has Come. The Primary Cilium
    I. Introduction
    II. Technical Issues in the Discovery and Study of Primary Cilia
    III. Origins: 1844-1910
    IV. The Henneguy-Lenhossek Hypothesis and the Role of Primary Cilia
    V. The Middle Period (1910-1935)
    VI. Origin of Photoreceptor Connecting Cilia from Primary Cilia
    VII. The Dark Ages of Primary Cilia (1935-1955)
    III. The 1950s and 1960s: Transmission Electron Microscopy and the "Renaissance" of the Primary Cilium
    IX. Primary Cilia and the Cell Cycle
    X. The Golden Age of Primary Cilia (2000-Onward)
    XI. Conclusions: The Morals of this Story
    References
    2. Origin of the Cilium: Novel Approaches to Examine a Centriolar Evolution Hypothesis
    I. Introduction
    II. Methods
    III. Viral Relatives of cnRNAs and Other Centriolar and Centrosomal Components
    IV. Conclusions
    References
    SECTION II Cell Biology/Biochemistry
    3. Using quantitative PCR to Identify Kinesin-3 Genes that are Upregulated During Growth Arrest in Mouse NIH3T3 Cells
    I. Introduction
    II. Rationale
    III. Materials
    IV. Methods
    V. Results and Discussion
    VI. Summary
    References
    4. Methods for the Isolation of Sensory and Primary Cilia--An Overview
    I. Introduction
    II. Isolation of Olfactory Cilia
    III. Isolation of Rod Photoreceptor Cilia
    IV. Isolation of Primary Cilia
    References
    5. Isolation of Primary Cilia for Morphological Analysis
    I. Introduction
    II. Materials and Instrumentations
    III. Methods
    IV. Results and Discussion
    V. Summary
    References
    6. Analyzing Primary Cilia by Multiphoton Microscopy
    I. Introduction
    II. Materials and Solutions
    III. Methods
    IV. Results and Discussion
    References
    7. Primary Cilia and the Cell Cycle
    I. Introduction
    II. Signaling Systems Regulating Ciliary Protusion and Resorption
    III. Methods
    References
    SECTION III Function
    8. Utilization of Conditional Alleles to Study the Role of the Primary Cilium in Obesity
    I. Introduction
    II. CNS Mechanisms of Energy Balance
    III. Human Ciliopathies and Obesity
    IV. Cilia Mutant Mouse Models of Obesity
    V. Neuronal Cilia
    VI. Strategies and Methods of Analyzing Obesity in Cilia Mouse Models
    References
    9. Using Nucleofection of siRNA Constructs for Knockdown of Primary Cilia in P19.CL6 Cancer Stem Cell Differentiation into Cardiomyocytes
    I. Introduction
    II. Rationale
    III. Materials
    IV. Methods
    V. Results and Discussion
    VI. Summary
    References
    10. The Primary Cilium as a Hedgehog Signal Transduction Machine
    I. Introduction
    II. Summary
    References
    11. Detecting the Surface Localization and Cytoplasmic Cleavage of Membrane-Bound Proteins
    I. Introduction
    II. Assay Rationale and History
    III. Materials
    IV. Methods
    V. Discussion
    VI. Summary
    References
    12. Assay for In Vitro Budding of Ciliary-Targeted Rhodopsin Transport Carriers
    I. Introduction
    II. Methods
    III. Materials
    IV. Results and Discussion
    V. Summary
    References
    13. Immunoelectron Microscopy of Vesicle Transport to the Primary Cilium of Photoreceptor Cells
    I. Introduction and Rationale
    II. Materials
    III. Methods and Procedures
    IV. Results and Discussion
    References
    14. Polycystic Kidney Disease, Cilia, and Planar Polarity
    I. Cystic Kidney Diseases
    II. The Primary Cilium and Cystic Kidney Diseases
    III. Explanatory Models for Cystic Kidney Diseases
    IV. Final Remarks
    References
    15. Constructing and Deconstructing Roles for the Primary Cilium in Tissue Architecture and Cancer
    I. Introduction
    II. Sensory Modalities
    III. Flow and Mechanosensation
    IV. Low-Abundance Ligands
    V. Concentration Shifts
    VI. Ciliary Receptors
    VII. Growth Factor Receptors
    VIII. Morphogens
    IX. Hormone and G-Protein-Coupled Receptors
    X. Tissue Homeostasis and Architecture
    XI. Cancer
    XII. Summary
    References
    SECTION IV Posttranslational Modifications
    16. Polyglutamylation and the fleer Gene
    I. Introduction
    II. Significance of Polyglutamylation
    III. Biochemistry of Tubulin Glutamylation
    IV. Functional Significance of Tubulin Glutamylase
    V. The Zebrafish fleer Mutant as a Paradigm for Analysis of Cilia Tubulin Glutamylation
    VI. Situs Inversus in fleer is Caused by Motility and Length Defects of Kupffer's Vesicle Cilia
    VII. Pronephric Cysts in fleer Result from Defects of Cilia Motility and Length
    VIII. Olfactory Placode Cilia
    IX. Abnormal Otolith Numbers are Caused by Defects of Motile Otic Placode Cilia
    X. Retinal Degeneration in fleer Results from Defects of the Connecting Cilium in Photoreceptors
    XI. fleer-Like Phenotypes in Other Species
    XII. Methods
    References
    17. Regulation of Cilia assembly, Disassembly, and Length by Protein Phosphorylation
    I. Introduction
    II. Protein Phosphorylation in Cilia Assembly
    III. Protein Phosphorylation in Cilia Disassembly
    IV. Protein Phosphorylation in Cilia Length Control
    V. Conclusion
    References
    18. Posttranslational Protein Modifications in Cilia and Flagella
    I. Introduction
    II. Posttranslational Protein Modification in Cilia and Flagella
    III. Methods for the Detection of PTM in Cilia and Flagella using the Electron Microscope
    References