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基本説明
The first in a three-part series on cilia. Contents: RNAi approaches to axonemal motor function in typanosomes; Homologous recombination to knock out dynein genes in Tetrahymena; and more.
Full Description
Along with its companion volume on intraflagellar transport, this book provides researchers with a comprehensive and up-to-date source of methods for the analysis cilia and flagella, focusing primarily on approaches that have been devised or significantly extended since the last volume of Methods in Cell Biology on this topic (volume 47, 1995). Edited by Stephen M. King and Gregory J. Pazour, the newest installment of this highly acclaimed serial will serve as an essential addition to the study of cilia and flagella.
Contents
1. RNAi approaches to axonemal motor function in trypanosomes (Hill)
2. Homologous recombination to knock out dynein genes in Tetrahymena (Gaertig)
3. RNAi in Chlamydomonas (Cerutti)
4. Planarians as a model system for analysis of ciliary assembly/motility (Rompolas)
5. Role of nucleotides in dynein function (Shingyoji)
6. Protein modification to probe dynein interactions + purification and identification of crosslinked dynein products (Sakato)
7. Analysis of redox-sensitive dynein components (Wakabayashi)
8. Purification of dyneins from Chlamydomonas and analysis of dynein-dynein linkers (Kamiya/King)
9. Kinase/phosphatase-mediated control of dynein function (Sale)
10. Central pair MT complex and associated kinesins (Mitchell)
11. Calcium regulation of axonemal function In vitro motility assays (Smith)
12. Identification and analysis of dynein regulatory complex components (Porter)
13. Isolation and analysis of radial spoke proteins (Yang)
14. Purification of dyneins from other model organisms: Ciona, sea urchin, fish (Inaba)
15. Rescue of mutant phenotypes by protein electroporation (Kamiya Lab)
16. Tubulin/Dynein interactions (Raff)
17. Measurement of beat frequency and swimming velocity Cell model reactivation in vitro assays of dynein function (Kamiya)
18. Live imaging of ependymal cilia (Lechtreck/Witman)
19. Immunogold labeling of axonemal components in situ + flat embedding (Geimer)
20. CryoEM approaches to axonemal organization (Nicastro)
21. CryoEM of dynein-microtubule complexes (Oda/Kikkawa)
22. Bioinformatic approaches to dynein HC classification (Yagi)
23. Bioinformatics of non-motor dynein components (Asai)
24. Biophysical approaches to dynein mechanism (Sutoh)
25. Chlamydomonas flagellar beat analysis (Foster)
26. Biophysical measurements of motor function (step size/force production) SAXS of axonemes (Toba/Oiwa)
Cryo-Electron Microscope Tomography to Study Axonemal
Organization
Daniela Nicastro
2. Electron Microscopic Imaging and Analysis of Isolated
Dynein Particles
Anthony J. Roberts and Stan A. Burgess
3. Immunogold Labeling of Flagellar Components In Situ
Stefan Geimer
4. Scanning Electron Microscopy to Examine Cells and Organs
Jovenal T. SanAgustin, John A. Follit, Gregory Hendricks, and Gregory J. Pazour
5. X-ray Fiber Diffraction Studies on Flagellar Axonemes
Kazuhiro Oiwa, Shinji Kamimura, and Hiroyuki Iwamoto
6. Markers for Neuronal Cilia
Jacqueline S. Domire and Kirk Mykytyn
7. Immunofluorescence Staining of Ciliated Respiratory Epithelial Cells
Heymut Omran and Niki T. Loges
8. Immunoprecipitation to Examine Protein Complexes
Gregory J. Pazour
9. Tandem Affinity Purification of Ciliopathy-Associated Protein
Complexes
Karsten Boldt, Jeroen van Reeuwijk, Christian Johannes Gloeckner,
Marius Ueffing, and Ronald Roepman
10. Crosslinking Methods: Purification and Analysis of Crosslinked Dynein Products
Miho Sakato
11. Analysis of the Ciliary/Flagellar Beating of Chlamydomonas
Kenneth W. Foster
12. Assays of Cell and Axonemal Motility in Chlamydomonas reinhardtii
Ritsu Kamiya
13. High-Speed Digital Imaging of Ependymal Cilia in the Murine Brain
Karl-Ferdinand Lechtreck, Michael J. Sanderson, and George B. Witman
14. Observation of Nodal Cilia Movement and Measurement of Nodal Flow
Yasushi Okada and Nobutaka Hirokawa
15. Modification of Mouse Nodal Flow by Applying Artificial Flow
Shigenori Nonaka
16. Measuring Cilium-Induced Ca2þ Increases in Cultured Renal Epithelia
Helle A. Praetorius
