Description

This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers cilia and includes chapters on such topics as methods for studying ciliary polarity in Xenopus, analysis of signaling pathways in mammalian spermatozoa, and biochemical and physiological analysis of axonemal dyneins.

Key Features

  • Continues the legacy of this premier serial with quality chapters authored by leaders in the field
  • Covers cilia
  • Contains chapters on such topics as methods for studying ciliary polarity in Xenopus,
    analysis of signaling pathways in mammalian spermatozoa, and biochemical and physiological analysis of axonemal dyneins

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

Contributors

Preface

Methods in Enzymology

Chapter One. Non-Motile Primary Cilia as Fluid Shear Stress Mechanosensors

1 Introduction

2 Identification of Primary Cilium

3 Mechanosensory Cilia

4 Summary

Acknowledgments

References

Chapter Two. Biomechanical Measurement of Kinocilium

1 Introduction

2 Sample Preparation and Measurement

3 Determining Mechanical Properties from Experimental Measurements

4 Method Discussion

Acknowledgments

References

Chapter Three. Analysis of Primary Cilia in Directional Cell Migration in Fibroblasts

1 Introduction

2 Localization of Primary Cilia by Immunofluorescence Microscopy

3 Cell Migration and Orientation of Primary Cilia in Scratch Assays

4 Cell Migration and Ciliary Signaling in Micropipette Assays

5 Summary

Acknowledgments

References

Chapter Four. Sperm Patch-Clamp

1 Introduction

2 Equipment

3 Isolation of Spermatozoa for Patch-Clamp Experiments

4 Patch-Clamp Recording

5 Potential Pitfalls Associated with Sperm Patch Clamp

6 Conclusions

Acknowledgments

References

Chapter Five. Laser Trap Measurements of Flagellar Membrane Motility

1 Introduction

2 Laser Trap Overview

3 Measuring Flagellar Membrane Transport

Acknowledgments

References

Chapter Six. Probing Ciliogenesis Using Micropatterned Substrates

1 Introduction

2 Micropatterned Substrates Fabrication

3 Cells Plating on Micropatterned Slides

4 Cell Fixation and Immunofluorescence

5 Automated Image Acquisition

6 Automated Image Analysis

Acknowledgments

References

Chapter Seven. Fluorescent Imaging of Drosophila melanogaster Sperm in the Reproductive Tract

1 Introduction

2 Ima

Details

No. of pages:
498
Language:
English
Copyright:
© 2013
Published:
Imprint:
Academic Press
Electronic ISBN:
9780123984852
Print ISBN:
9780123979445

About the serial-volume-editor

Wallace F. Marshall

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.