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