Leading drosophilists describe in step-by-step detail all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. The chapters emphasize specimen preparation (from dissection to mounting) and cover both polytene and mitotic/meiotic chromosomes in depth. Each fully tested and readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls. A cutting-edge FISH and immunolocalization technique will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression.
The Chromosomes of Drosophila melanogaster Daryl S. Henderson Spermatogenesis: Analysis of Meiosis and Morphogenesis Helen White-Cooper Time-Lapse Imaging of Male Meiosis by Phase-Contrast and Fluorescence Microscopy Elena Rebollo and Cayetano González Immunocytological Analysis of Oogenesis Endre Máthé Cytological Analysis of Oogenesis Seppo Nokkala and Christina Nokkala Polytene Chromosomes From Ovarian Nurse Cells of Drosophila melanogaster otu Mutants Dmitry E. Koryakov, Natalia I. Mal'ceva, Robert C. King, and Igor F. Zhimulev Feulgen-DNA Cytophotometry for Estimating C Values Ellen M. Rasch Fluorescent BrdU Labeling and Nuclear Flow Sorting of the Drosophila Ovary Brian R. Calvi and Mary A. Lilly Studying Nuclear Organization in Embryos Using Antibody Tools Kristen M. Johansen and Jørgen Johansen Whole-Mount Fluorescence In Situ Hybridization to Chromosomes of Embryos Daryl S. Henderson Orcein Staining and the Identification of Polytene Chromosomes John Tonzetich Salivary Chromosome Analysis of Aberrations Adelaide T. C. Carpenter In Situ Hybridization to Polytene Chromosomes Robert D. C. Saunders Combined Immunostaining and FISH Analysis of Polytene Chromosomes Sergey Lavrov, Jérôme Déjardin, and Giacomo Cavalli Electron Microscopy of Polytene Chromosomes Valery F. Semeshin, Elena S. Belyaeva, Victor V. Shloma, and Igor F. Zhimulev Analysis of Mitosis in Squash Preparations of Larval Brains: Orcein, Giemsa, Hoechst 33258, DAPI, Quinacrine, and N-Banding Laura Fanti and Sergio Pimpinelli Analyzing Chromosome Function by High Frequency Formation of Dicentric Chromosomes In Vivo Kami Ahmad and Kent G. Golic Analysis of Chromosomes of the Larval CNS by FISH and BrdU Labeling Amy K.Csink Immunostaining of Squash Preparations of Chromosomes of Larval Brains Laura Fanti and Sergio Pimpinelli Visualizing Mitosis in Whole-Mount Larval Brains Daryl S. Henderson Immunostaining of Whole-Mount Imaginal Discs Brigitte de Saint Phalle Wing Somatic Mutation and Recombination Test Heloísa Helena Rodrigues de Andrade, Maria Luíza Reguly, and Mauricio Lehmann Analysis of Histoblasts Mekkara Mandaravally Madhavan and Kornath Madhavan Visualizing Apoptosis Kimberly McCall, Jason S. Baum, Kristen Cullen, and Jeanne S. Peterson RNAi in Cultured Drosophila Cells Ling-Rong Kao and Timothy L. Megraw Index
Probably no single experimental organism has contributed as much to our understanding of the structure, function, and mechanism of transmission of eukaryotic chromosomes as has the fruit fly, Drosophila melanogaster. In Drosophila Cytogenetics Protocols, leading drosophilists describe, in step-by-step detail, all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. General areas covered include analyses of nuclei and chromosomes in spermatogenesis and oogenesis, fluorescence-activated cell sorting of ovarian follicle cells, analyses of nuclear division cycles in embryos, and analyses of programmed cell death. In addition, there are conventional light-, electron-, and immunofluorescence-microscopic analyses of polytene chromosomes; cytological analyses of larval brains, imaginal discs, and histoblasts; and nonfluorescent and fluorescent in situ hybridization (FISH) to both polytene and mitotic chromosomes. A cutting-edge combined FISH and immunolocalization protocol will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression. Each readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls.
Readily accessible and highly practical, Drosophila Cytogenetics Protocols provides a comprehensive cytogenetics laboratory manual for Drosophila investigators, one that is suitable not only for novices, but also highly informative for seasoned investigators.
...this volume will be worth returning to for years to come...the spirit of this volume is not that it will get you through, but that it will make you comfortable with the technology and put you in touch with the art form...a valuable and stimulating source of essential technical information for all experimental 'drosophilists'. - Nature Genetics
Inhaltsverzeichnis
The Chromosomes of Drosophila melanogaster
Daryl S. Henderson
Spermatogenesis: Analysis of Meiosis and Morphogenesis
Helen White-Cooper
Time-Lapse Imaging of Male Meiosis by Phase-Contrast and Fluorescence Microscopy
Elena Rebollo and Cayetano González
Immunocytological Analysis of Oogenesis
Endre Máthé
Cytological Analysis of Oogenesis
Seppo Nokkala and Christina Nokkala
Polytene Chromosomes From Ovarian Nurse Cells of Drosophila melanogaster otu Mutants
Dmitry E. Koryakov, Natalia I. Mal'ceva, Robert C. King, and Igor F. Zhimulev
Feulgen-DNA Cytophotometry for Estimating C Values
Ellen M. Rasch
Fluorescent BrdU Labeling and Nuclear Flow Sorting of the Drosophila Ovary
Brian R. Calvi and Mary A. Lilly
Studying Nuclear Organization in Embryos Using Antibody Tools
Kristen M. Johansen and Jørgen Johansen
Whole-Mount Fluorescence In Situ Hybridization to Chromosomes of Embryos
Daryl S. Henderson
Orcein Staining and the Identification of Polytene Chromosomes
John Tonzetich
Salivary Chromosome Analysis of Aberrations
Adelaide T. C. Carpenter
In Situ Hybridization to Polytene Chromosomes
Robert D. C. Saunders
Combined Immunostaining and FISH Analysis of Polytene Chromosomes
Sergey Lavrov, Jérôme Déjardin, and Giacomo Cavalli
Electron Microscopy of Polytene Chromosomes
Valery F. Semeshin, Elena S. Belyaeva, Victor V. Shloma, and Igor F. Zhimulev
Analysis of Mitosis in Squash Preparations of Larval Brains: Orcein, Giemsa, Hoechst 33258, DAPI, Quinacrine, and N-Banding
Laura Fanti and Sergio Pimpinelli
Analyzing Chromosome Function by High Frequency Formation of Dicentric Chromosomes In Vivo
Kami Ahmad and Kent G. Golic
Analysis of Chromosomes of the Larval CNS by FISH and BrdU Labeling
Amy K.Csink
Immunostaining of Squash Preparations of Chromosomes of Larval Brains
Laura Fanti and Sergio Pimpinelli
Visualizing Mitosis in Whole-Mount Larval Brains
Daryl S. Henderson
Immunostaining of Whole-Mount Imaginal Discs
Brigitte de Saint Phalle
Wing Somatic Mutation and Recombination Test
Heloísa Helena Rodrigues de Andrade, Maria Luíza Reguly, and Mauricio Lehmann
Analysis of Histoblasts
Mekkara Mandaravally Madhavan and Kornath Madhavan
Visualizing Apoptosis
Kimberly McCall, Jason S. Baum, Kristen Cullen, and Jeanne S. Peterson
RNAi in Cultured Drosophila Cells
Ling-Rong Kao and Timothy L. Megraw
Index
Klappentext
Leading drosophilists describe in step-by-step detail all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. The chapters emphasize specimen preparation (from dissection to mounting) and cover both polytene and mitotic/meiotic chromosomes in depth. Each fully tested and readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls. A cutting-edge FISH and immunolocalization technique will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression.
Includes supplementary material: sn.pub/extras
