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Cilia and Flagella presents protocols accessible to all individuals working with eukaryotic cilia and flagella. These recipes delineate laboratory methods and reagents, as well as critical steps and pitfalls of the procedures. The volume covers the roles of cilia and flagella in cell assembly and motility, the cell cycle, cell-cell recognition and other sensory functions, as well as human diseases and disorders. Students, researchers, professors, and clinicians should find the book's combination of "classic" and ...
Cilia and Flagella presents protocols accessible to all individuals working with eukaryotic cilia and flagella. These recipes delineate laboratory methods and reagents, as well as critical steps and pitfalls of the procedures. The volume covers the roles of cilia and flagella in cell assembly and motility, the cell cycle, cell-cell recognition and other sensory functions, as well as human diseases and disorders. Students, researchers, professors, and clinicians should find the book's combination of "classic" and innovative techniques essential to the study of cilia and flagella.
• A complete guide containing more than 80 concise technical chapters friendly to both the novice and experienced researcher
• Covers protocols for cilia and flagella across systems and species from Chlamydomonas and Euglena to mammals
• Both classic and state-of-the-art methods readily adaptable across model systems, and designed to last the test of time, including microscopy, electrophoresis, and PCR
• Relevant to clinicians interested in respiratory disease, male infertility, and other syndromes, who need to learn biochemical, molecular, and genetic approaches to studying cilia, flagella, and related structures
Audience: Graduate students, technicians, clinicians, and experienced researchers interested in the study of cilia and flagella. Cell, developmental, and molecular biologists, biochemists, geneticists, physiologists, neurobiologists, medical researchers, and biotechnologists.
Isolation of Cilia and Flagella:
P.A. Lefebvre, Flagellar Amputation and Regeneration in Chlamydomonas.
S.M. King, Large-Scale Isolation of Chlamydomonas Flagella.
W.L. Dentler, Isolation of Cilia from Tetrahymena thermophila.
D.L. Nelson, Preparation of Cilia and Subciliary Fractions from Paramecium.
H.M. Ngo and G.B. Bouck, Isolation of Euglena Flagella.
J.T.S. Agustin and G.B.Witman, Isolation of Ram Sperm Flagella.
R.E. Stephens, Isolation of Molluscan Gill Cilia, Sperm Flagella, and Axonemes.
L.H. Cheung, Isolation of Flagella and Their Membranes from Sea Urchin Spermatozoa.
C. Saudrais and J.-L. Gatti, Isolation of Fish Sperm Flagella.
Culture and Isolation of Ciliated Epithelial Cells and Cilia:
L.E. Ostrowski, S.H. Randell, A.B. Clark, T.E. Gray, and P. Nettesheim, Ciliogenesis of Rat Tracheal Epithelial Cells in Vitro.
E.R. Dirksen, J.A. Felix, and M.J. Sanderson, Preparation of Explant and Organ Cultures and Single Cells from Airway Epithelium.
B. Chailley and E. Boisvieux-Ulrich, Induction of Ciliogenesis in Oviduct Epithelium.
K. Pagh-Roehl and B. Burnside, Preparation of Teleost Rod Inner and Outer Segments.
A.T. Hastie, Isolation of Respiratory Cilia.
G. Piperno, Electrophoretic Separation of Dynein Heavy Chains.
G. Piperno, Two-Dimensional Separation of Axonemal Proteins.
T. Otter, Analysis of Flagellar Calcium-Binding Proteins.
R.A. Bloodgood and N.L. Salomonsky, Phosphorylation of Chlamydomonas Flagellar Proteins.
S.-J.Chen, T.K. Rosiere, and G.B. Bouck, In Situ Glycosylation of Flagellar Lipids.
J.T. San Agustin and G.B. Witman, Detection of Flagellar Protein Kinases on Polyvinylidene Fluoride Membranes Following Sodium Dodecyl Sulfate(Polyacrylamide Gel Electrophoresis.
S.M. King, Measurement of ATPase Activity Using
O. Kagami and R. Kamiya, Nonradioactive Method for ATPase Assays.
K.A. Johnson, Immunoelectron Microscopy.
M.A. Sanders and J.L. Salisbury, Immunofluorescence Microscopy of Cilia and Flagella.
S.P. Marchese-Ragona and J.S. Wall, Scanning Transmission Electron Microscopic Analysis of the Isolated Dynein ATPase.
S.P. Marchese-Ragona, High-Resolution Negative Staining of the Isolated Dynein ATPase.
E. O'Toole, D. Mastronarde, J.R. McIntosh, and M.E. Porter, Computer-Assisted Analysis of Flagellar Structure.
H.J. Hoops, Preparation of Cilia and Flagella for Thin-Section Transmission Electron Microscope Analysis.
D.M. Phillips, Fixation of Mammalian Spermatozoa for Electron Microscopy.
Motors and Motion Analysis:
C.J. Horst and G.B. Witman, Reactivation of Chlamydomonas Cell Models.
Y. Naitoh, Reactivation of Extracted Paramecium Models.
R. Hard, Isolation and Reactivation of Newt Lung Respiratory Cilia.
C.J. Brokaw, Reactivation of Motility of Demembranated Sea Urchin Sperm Flagella.
S. Ishijima, High-Speed Video Microscopy of Flagella and Cilia.
S. Ishijima, Micromanipulation of Sperm and Other Ciliated or Flagellated Single Cells.
J.T. San Agustin and G.B. Witman, Preparation and Reactivation of Demembranated, Cytosol-Free Ram Spermatozoa.
W.S. Sale and D.R. Howard, Microscopic Assays of Flagellar Dynein Activity.
K.G. Kozminski, High-Resolution Imaging of Flagella.
R.A. Bloodgood, Flagellar Surface Motility: Gliding and Microsphere Movements.
A.G. Moss, G.J. Pazour, and G.B. Witman, Assay of Chlamydomonas Phototaxis.
M.J. Sanderson and E.R. Dirksen, Quantification of Ciliary Beat Frequency and Metachrony by High-Speed Digital Video.
N.E. Williams and J.E. Honts, Isolation and Fractionation of the Tetrahymena Cytoskeleton and Oral Apparatus.
B.E. Taillon and J.W. Jarvik, Release of the Cytoskeleton and Flagellar Apparatus from Chlamydomonas.
A. Bremerich, K.-F. Lechtreck, and M. Melkonian, Purification of SF-Assemblin.
S.K. Dutcher, Purification of Basal Bodies and Basal Body Complexes from Chlamydomonas reinhardtii.
K.-F. Lechtreck and M. Melkonian, Preparation and Reactivation of Spermatozopsis Cytoskeletons.
A.T. Baron, R. Errabolu, J. Dinusson, and J.L. Salisbury, Centrin-Based Contractile Fibers: Chromatographic Purification of Centrin.
Flagellar Fractionation and Membrane Characterization:
H.M. Ngo and G.B. Bouck, Isolation of Flagellar Paraxonemal Rod Proteins.
R.E. Stephens, Preparation of Ciliary and Flagellar Remnants.
R. Kuriyama and R.W. Linck, In Vitro Polymerization of Tubulin from Echinoderm Sperm Flagellar Microtubules.
M.A. Pirner and R.W. Linck, Methods for the Isolation of Tektins and Sarkosyl-Insoluble Protofilament Ribbons.
G. Piperno, Isolation of Radial Spoke Heads from Chlamydomonas Axonemes.
M. Brito and L.O. Burzio, Isolation of the Dense Fibers of Mammalian Sperm Flagella.
M. Brito and L.O. Burzio, Isolation of the Fibrous Sheath of Mammalian Sperm Flagella.
W.L. Dentler, Isolation and Fractionation of Ciliary Membranes from Tetrahymena.
L.H. Cheung, Measurement of Membrane Potential and Na+ and H+ Transport in Isolated Sea Urchin Sperm Flagella and Their Membrane Vesicles.
W.L. Dentler, Nonradioactive Methods for Labeling and Identifying Membrane Surface Proteins.
R. Wright, Fluorescence Labeling of Flagellar Membranes.
H. Machemer, Electrophysiology of Ciliates.
M. Bernstein, Isolation of Chlamydomonas Mastigonemes.
R.E. Stephens, Ciliary Membrane Tubulin: Isolation and Fractionation.
L.J. Dangott, Peptide Receptors in Sea Urchin Sperm Flagella.
Assay and Isolation of Flagellar Enzymes:
G.E. Bracho and J.S. Tash, Assaying Protein Phosphatases in Sperm and Flagella.
Y. Zhang and W.J. Snell, Flagellar Adenylyl Cyclases in Chlamydomonas.
R.M. Tombes, Isolation and Characterization of Sea Urchin Flagellar Creatine Kinase.
G.L. Carlson and D.L. Nelson, Isolation and Characterization of Protein Kinases of Paramecium Cilia.
D.R. Howard and W.S. Sale, Isolation of Inner- and Outer-Arm Dyneins.
O. Kagami and R. Kamiya, Separation of Dynein Species by High-Pressure Liquid Chromatography.
E.F. Smith, Reconstitution of Dynein Arms in Vitro.
T. Shimizu, Inhibitors of the Dynein ATPase and Ciliary or Flagellar Motility.
S.M. King, Vanadate-Mediated Photolysis of Dynein Heavy Chains.
C.K. Omoto, Modified Nucleotides as Probes for Dynein.
Genetics and Molecular Tools for the Analysis of Cilia and Flagella:
J.A.E. Nelson and P.A. Lefebvre, Transformation of Chlamydomonas reinhardtii.
L.-W. Tam and P.A. Lefebvre, Insertional Mutagenesis and Isolation of Tagged Genes in Chlamydomonas.
C.D. Silflow, P. Kathir, and P.A. Lefebvre, Molecular Mapping of Genes for Flagellar Proteins in Chlamydomonas.
S.K. Dutcher, Mating and Tetrad Analysis in Chlamydomonas reinhardtii.
R. Kamiya, Strategies for the Isolation of the Flagellar Motility and Assembly Mutants in Chlamydomonas.
D.R. Diener, Epitope Tagging of Flagellar Proteins.
L.R. Keller, Enrichment of mRNA Encoding Flagellar Proteins.
J. Gaertig and M.A. Gorovsky, DNA-Mediated Transformation in Tetrahymena.
D.G. Pennock and M.A. Gorovsky, Strategies for the Isolation of Ciliary Motility and Assembly Mutants inTetrahymena.
D.J. Asai and P.S. Criswell, Identification of New Dynein Heavy-Chain Genes by RNA-Directed Polymerase Chain Reaction.
Volumes in Series.