Cilia: Model Organisms and Intraflagellar Transport

Cilia are highly conserved organelles that serve motile functions, sensory functions, or both. These organelles power cell movement, generate fluid flow in various organs, act as sensors of the extracellular environment and have been modified for various specialized tasks such as light reception and smell. Defects in these ubiquitous organelles lead to a broad array of human genetic disorders that range from polycystic kidney disease, retinal degeneration, epilepsy and infertility to developmental defects such as situs inversus and polydactyly. This volume is the third in a three-part series on cilia that focuses on the use of model organisms to gain insight into ciliary function and on the process of intraflagellar transport that is essential for the assembly and maintenance of ciliary structures.

* Includes both classic and state-of-the-art methods readily adaptable across model systems, and designed to last the test of time * Covers forward and reverse genetic analysis of IFT and biochemical methods to define the role of IFT components * Methods presented cover molecular, genetic, and biochemical approaches to ciliary function in model organisms

1. Manipulating Ciliary Protein-Encoding Genes in Tetrahymena thermophila

Drashti Dave, Dorota Wloga, and Jacek Gaertig

2. Approaches for Functional Analysis of Flagellar Proteins in African Trypanosomes

Michael Oberholzer, Miguel A. Lopez, Katherine S. Ralston, and Kent L. Hill

3. Tools for Analyzing Intraflagellar Transport in Trypanosomes

Daria Julkowska and Philippe Bastin

4. Schmidtea mediterranea: A Model System for Analysis of Motile Cilia

Panteleimon Rompolas, Ramila S. Patel-King, and Stephen M. King

5. Targeted Gene Silencing by RNA Interference in Chlamydomonas

Eun-Jeong Kim and Heriberto Cerutti

6. Analysis of Cargo Transport by IFT and GFP Imaging of IFT in Chlamydomonas

Dennis Diener

7. Genetic and Phenotypic Analysis of Flagellar Assembly Mutants

in Chlamydomonas reinhardtii

Carlo Iomini, Jacob E. Till, and Susan K. Dutcher

8. Recording and Analyzing IFT in Chlamydomonas Flagella

William Dentler, Kristyn VanderWaal, and Mary E Porter

9. Total Internal Reflection Fluorescence (TIRF) Microscopy

of Chlamydomonas Flagella

Benjamin D. Engel, Karl-Ferdinand Lechtreck, Tsuyoshi Sakai, Mitsuo Ikebe, George B. Witman, and

Wallace F. Marshall

10. Purification of IFT Particle Proteins and Preparation of Recombinant

Proteins for Structural and Functional Analysis

Robert H. Behal, Ewelina Betleja, and Douglas G. Cole

11. Studying Cilia in Zebrafish

Iain Drummond

12. Analysis of IFT Kinesins in Developing Zebrafish Cone Photoreceptor

Sensory Cilia

Christine Insinna, Katherine Luby-Phelps, Brian A. Link, and Joseph C. Besharse

13. Analysis of Intraflagellar Transport in C. elegans Sensory Cilia

Limin Hao, Seyda Acar, James Evans, Guangshuo Ou, and Jonathan M. Scholey

14. Functional Genomics of Intraflagellar Transport-Associated Proteins in C. elegans

Peter N. Inglis, Oliver E. Blacque, and Michel R. Leroux

15. Generating Conditional Mutants to Analyze Ciliary Functions:

The Use of Cre–Lox Technology to Disrupt Cilia in Specific Organs

Amber K. O’Connor, Robert A. Kesterson, and Bradley K. Yoder

16. Imaging Intraflagellar Transport in Mammalian Primary Cilia

Tatiana Y. Besschetnova, Barnali Roy, and Jagesh V. Shah

17. Analysis of Hedgehog Signaling in Mouse Intraflagellar Transport Mutants

Hyuk W. Ko, Aimin Liu, and Jonathan T. Eggenschwiler