Methods in Cilia and Flagella - 1st Edition - ISBN: 9780128024515, 9780128026403

Methods in Cilia and Flagella, Volume 127

1st Edition

Serial Volume Editors: Renata Basto Wallace F. Marshall
eBook ISBN: 9780128026403
Hardcover ISBN: 9780128024515
Imprint: Academic Press
Published Date: 25th March 2015
Page Count: 614
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Table of Contents

  • Series Editors
  • Contributors
  • Preface
  • Chapter 1. Imaging centrosomes and cilia in the mouse kidney
    • Introduction
    • 1. Methods
    • Conclusion
  • Chapter 2. Ependymal cell differentiation, from monociliated to multiciliated cells
    • Introduction
    • 1. Methods
    • Conclusion
  • Chapter 3. Observing planar cell polarity in multiciliated mouse airway epithelial cells
    • Introduction
    • 1. Acquisition or Generation of Multiciliated Airway Epithelial Cells
    • 2. Assessment of Ultrastructural Planar Polarity of Motile Cilia
    • 3. Localization of PCP Signaling Pathway Proteins in Multiciliated Airway Epithelial Cells
    • Conclusions
  • Chapter 4. Imaging and analyzing primary cilia in cardiac cells
    • Introduction
    • 1. Imaging Cilia In situ, in Fixed Hearts
    • 2. Imaging and Manipulating Cilia in Primary Cultures of Cardiac Cells
    • 3. Quantitative Analysis of Cilium Orientation
    • Conclusion
  • Chapter 5. Cilia in photoreceptors
    • Introduction
    • 1. Methods
    • Conclusions
  • Chapter 6. Analysis of primary cilia in the developing mouse brain
    • Introduction
    • 2. Primary Cilia in the Developing Neocortex
    • 3. Material
    • 4. Methods
    • Conclusion
  • Chapter 7. In vivo investigation of cilia structure and function using Xenopus
    • Introduction
    • 1. A Brief Introduction to Xenopus Husbandry and Manipulation
    • 2. Imaging of Cilia in Fixed Xenopus Embryos and Tissues
    • 3. Methods for Live Cell Analysis of Cilia in Xenopus
    • 4. Mounting Xenopus Embryos
    • 5. Image Acquisition
    • 6. Quantitative Analysis of Ciliary Properties
    • Conclusion
  • Chapter 8. A quantitative approach to study endothelial cilia bending stiffness during blood flow mechanodetection in vivo
    • Introduction
    • 1. The Challenge of Measuring Ciliary Mechanics
    • 2. Live Imaging
    • 3. Image Processing
    • 4. Bending Rigidity Estimation through Modeling
    • 5. Material
    • Conclusion
  • Chapter 9. Quantitative description of fluid flows produced by left–right cilia in zebrafish
    • Introduction
    • 1. Methods
    • 2. Application
    • Conclusion
  • Chapter 10. Efficient live fluorescence imaging of intraflagellar transport in mammalian primary cilia
    • Introduction
    • 1. Preparation of Cells for IFT Imaging
    • 2. Live Imaging of IFT
    • 3. Image Processing and Data Analysis
    • 4. Discussion
  • Chapter 11. Analysis of soluble protein entry into primary cilia using semipermeabilized cells
    • Introduction
    • 1. Methods
    • 2. Conclusions and Future Outlook
  • Chapter 12. Methods for imaging individual cilia in living echinoid embryos
    • Introduction
    • 1. Methods
    • Conclusion
  • Chapter 13. The planarian Schmidtea mediterranea as a model for studying motile cilia and multiciliated cells
    • Introduction
    • 1. Maintaining Planarians in the Lab
    • 2. RNA Interference
    • 3. Whole-Mount Immunofluorescence Staining
    • 4. Electron Microscopy
    • 5. Live Imaging
  • Chapter 14. Quantitative analysis of flagellar proteins in Drosophila sperm tails
    • Introduction
    • 1. Methods
    • Concluding Remarks
  • Chapter 15. Imaging cilia in Drosophila melanogaster
    • Introduction
    • 1. Immunodetection of Cilia
    • 2. Observation of Cilia Ultrastructure by Transmission Electron Microscopy
  • Chapter 16. Studying G protein-coupled receptors: immunoblotting, immunoprecipitation, phosphorylation, surface labeling, and cross-linking protocols
    • Introduction
    • 1. Reagents
    • 2. Protocol for Immunoblotting GPCRs
    • 3. Protocol for Immunoprecipitating GPCRs
    • 4. GPCR Labeling and Cross-Linking Studies
    • 5. Comments on Biochemical Assays Involving GPCRs and Multipass Transmembrane Proteins
    • Conclusions
  • Chapter 17. Image analysis of Caenorhabditis elegans ciliary transition zone structure, ultrastructure, molecular composition, and function
    • Introduction
    • 2. Caenorhabditis elegans Sensory Neuronal Cilia Model
    • 3. Methods
    • Concluding Remarks
  • Chapter 18. Genetic and genomic approaches to identify genes involved in flagellar assembly in Chlamydomonas reinhardtii
    • Introduction
    • 1. Identifying the Causative Genes Using Forward Genetics
    • 2. Identifying Gene Function Using Reverse Genetics
    • 3. Identifying Genes Using the RNA Transcriptome
    • 4. Identifying Genes Using Genetic Interactions
    • 5. Summary
  • Chapter 19. Axonemal motility in Chlamydomonas
    • Introduction
    • 1. Methods
  • Chapter 20. Posttranscriptional control over rapid development and ciliogenesis in Marsilea
    • Introduction
    • 1. The Development of Experimental Protocols in a Nonmodel System
    • 2. Centrin Translation is Necessary for Formation of the Blepharoplast
    • 3. Cell Fate in the Male Gametophyte of Marsilea: Only Two Types of Cells are Produced
    • 4. Spermidine Provides a Driving Force behind Rapid Development in the Male Gametophyte
    • 5. Spermidine Levels Change in the Gametophyte
    • 6. Stored mRNAs are Transcribed during the Natural Process of Spore Desiccation
    • 7. Mago Nashi and Speckle mRNA Dynamics
    • 8. Emerging Themes from the Transcriptome of the Rapidly Developing Male Gametophyte of Marsilea
    • 9. Many of the Stored Transcripts in the Spore Nucleus are Partially Processed Pre-mRNAs
    • 10. NudC, an Archetype IRT in the Gametophyte
    • 11. IRT Splicing as a Means to Regulate the Timing and Location of Translation
    • 12. Polyadenylation as a Late Regulator of Translation
    • Conclusions, Predictions, and a Working Model for the Control of Rapid Development
  • Chapter 21. Total internal reflection fluorescence microscopy of intraflagellar transport in Tetrahymena thermophila
    • Introduction
    • 1. TIRF Microscope Configuration and Data Analysis in Application to IFT in Tetrahymena
    • 2. Using a TIRF microscope to image Tetrahymena cilia
    • 3. Limitations
  • Chapter 22. Ciliary heterogeneity within a single cell: The Paramecium model
    • 1. The Paramecium Model
    • 2. Cilia in Paramecium Organization
    • 3. Staining the Cilia with Antitubulin Antibodies
    • 4. Protocols
    • 5. Conclusion
    • 6. Chemical Reagents and Buffers
  • Chapter 23. Imaging intraflagellar transport in trypanosomes
    • Introduction
    • 1. Producing Trypanosomes Expressing Fluorescent IFT Fusion Proteins
    • 2. Visualization of IFT and Acquisition of Time-Lapse Videos
    • 3. Quantification of IFT
    • Conclusion
  • Chapter 24. Scanning and three-dimensional electron microscopy methods for the study of Trypanosoma brucei and Leishmania mexicana flagella
    • Introduction
    • 1. Rationale
    • 2. Methods
    • 3. Materials
    • 4. Discussion
  • Volumes in Series
  • Index

Description

The goal of this book is to collect methods and protocols for studying cilia in a wide range of different cell types, so that researchers from many fields of biology can start exploring the role of cilia in their own system.

Key Features

  • Chapters are written by experts in the field
  • Cutting-edge material

Readership

Biological researchers at any level who want to learn methods for studying cilia in different cell types


Details

No. of pages:
614
Language:
English
Copyright:
© Academic Press 2015
Published:
Imprint:
Academic Press
eBook ISBN:
9780128026403
Hardcover ISBN:
9780128024515

About the Serial Volume Editors

Renata Basto Serial Volume Editor

Affiliations and Expertise

Cell Biology Department, CNRS, Institut Curie, France

Wallace F. Marshall Serial Volume Editor

Wallace Marshall is an electrical engineer by training, who became interested in biology out of a desire to understand how cells solve engineering problems, such as determining the size of organelles. He received his Ph.D. at UCSF with John Sedat, where he studied the diffusional of motion of interphase chromatin using live cell imaging and computational image analysis. He then trained as a postdoc with Joel Rosenbaum at Yale, where he began studying the mechanisms regulating the length of cilia and flagella. He is now Profess of Biochemistry at UCSF, where he lab continues to study the assembly and length regulation of cilia and flagella, as well as the mechanisms that regulate the size of other organelles. His work takes advantage of an integrated combination of methods including genetics, microscopy, and computational modeling, as well as a wide variety of model organisms including Chlamydomonas reinhardtii, Stentor coeruleus, yeast, flatworms, and mammalian cells.

Affiliations and Expertise

Department of Biochemistry & Biophysics, University of California San Fransisco at Mission Bay, USA