Circadian rhythms : methods and protocols / edited by Ezio Rosato. — Totowa, N.J. : Humana Press, c2007. -- (58.17/M592/v.362)

Contents

    Contents
    
    Preface v
    Contributors xi
    PART I. OVERWEWS
    1. Light， Photoreceptors， and Circadian Clocks 3
    2. Statistical Analysis of Biological Rhythm Data 29
    PART II. RHYTHMIC REAOOUTS
    3. Rhythmic Conidiation in Neurospora crassa 49
    4. Monitoring and Analyzing Drosophila Circadian Locomotor Activity 67
    5. Automated Video Image Analysis of Larval Zebrafish Locomotor Rhythms 83
    6. Locomotor Activity in Rodents 95
    7. Analysis of Circadian Leaf Movement Rhythms in Arabidopsis thaliana 103
    8. Detection of Rhythmic Bioluminescence From Luciferase Reporters in Cyanobacteria 115
    9. Analysis of Rhythmic Gene Expression in Adult Drosophila Using the Firefly Luciferase Reporter Gene 131
    10. Monitoring Circadian Rhythms in Arabidopsis thaliana Using Luciferase Reporter Genes 143
    Part III. Identification of CLOCK Genes
    11. Specialized Techniques for Site-Directed Mutagenesis in Cyanobacteria 155
    12. Novel Strategies for the Identification of Clock Genes in Neurospora With Insertional Mutagenesis 173
    13. Mutagenesis With Drosophila 187
    14. Mutagenesis in Arabidopsis 197
    15. Yeast Two-Hybrid Screening 207
    16. Microarrays: Quality Control and Hybridization Protocol 225
    17. Microarrays: Statistical Methods for Circadian Rhythms 245
    18. Identification of Clock Genes Using Difference Gel Electrophoresis 265
    Part IV. Gone Expression: RNA
    19. Isolation of Total RNA From Neurospora Mycelium 291
    20. RNA Extraction From Drosophila Heads 305
    21. Extraction of Plant RNA 309
    22. RNA Extraction From Mammalian Tissues 315
    23. Northern Analysis of Sense and Antisense frequency RNA in Neurospora crassa 329
    24. RNase Protection Assay 343
    25. Quantitative Polymerase Chain Reaction 349
    PART V. GENE EXPRESSION: PROTEINS
    26. Protein Extraction， Fractionation， and Purification From Cyanobacteria 365
    27. Protein Extraction From Drosophila Heads 375
    28. Plant Protein Extraction 379
    29. Protein Extraction From Mammalian Tissues 385
    30. Western Blotting 391
    31. Coimmunoprecipitation Assay 401
    32. In Vitro Phosphorylation and Kinase Assays in Neurospora crassa 407
    PART VI. IN VITRO SYSTEMS
    33. Basic Protocols for Drosophila S2 cell line: Maintenance and Transfection 415
    34. Coimmunoprecipitation on Drosophila Cells in Culture 423
    35. Basic Protocols for Zebrafish Cell Lines: Maintenance and Transfection 429
    36. Manipulation of Mammalian Cell Lines for Circadian Studies 443
    37. Reporter Assays 455
    38. Use of Firefly Luciferase Activity Assays to Monitor Circadian Molecular Rhythms In Vivo and In Vitro 465
    39. Suprachiasmatic Nucleus Cultures That Maintain Rhythmic Properties In Vitro 481
    PART VII. MICROSCOPY ANALYSIS
    40. RNA In Situ Hybridizations on Drosophila Whole Mounts 495
    41. In Situ Hybridization of Suprachiasmatic Nucleus Slices 513
    42. Immunohistochemistry in Drosophila: Sections and Whole Mounts 533
    43. Immunocytochemistry on Suprachiasmatic Nucleus Slices 549
    44. Immunofluorescence Analysis of Circadian Protein Dynamics in Cultured Mammalian Cells 561
    Index 569