Main description:

Our understanding of the molecular and cellular mechanisms that control skeletal muscle development, regeneration, and adaptive responses to activity has increased dramatically in recent years, fostered by innovative techniques and approaches that are either specifically designed or adapted for research in skeletal muscle biology. Myogenesis: Methods and Protocols presents detailed, step-by-step methods in the study of the molecular and cellular biology of skeletal muscle cells. Protocols from different model systems including mammalian, avian, zebrafish, and invertebrate skeletal muscle are included in this volume. Highlighted topics cover a wide range of interests and expertise including myogenic and stem cell isolation, investigation of models of exercise and disuse, viral vector delivery systems, calcium imaging, cell profiling, as well as protein-DNA and protein-protein interactions. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and authoritative, Myogenesis: Methods and Protocols serves as an invaluable, state-of-the-art resource for experienced and emerging scientists in basic research as well as clinical and regenerative medicine.

Contents:

Part I: Isolation and Analysis of Skeletal Muscle Precursor Cells

1. Isolation and Characterization of Human Fetal Myoblasts

Ariya D. Lapan and Emanuela Gussoni

2. Skeletal Muscle Satellite Cells: Background and Methods for Isolation and Analysis in a Primary Culture System

Maria Elena Danoviz and Zipora Yablonka-Reuveni

3. Isolation of Muscle Stem Cells by Fluorescence Activated Cell Sorting (FACS) Cytometry

Alessandra Pasut, Paul Oleynik, and Michael A. Rudnicki

4. Mouse and Human Mesoangioblasts: Isolation and Characterization from Adult Skeletal Muscles

Mattia Quattrocelli, Giacomo Palazzolo, Ilaria Perini, Stefania Crippa, Marco Cassano, and Maurilio Sampaolesi

5. Direct Electrical Stimulation of Myogenic Cultures for Analysis of Muscle Fiber Type Control

Eric J. Cavanaugh, Jennifer R. Crew, and Joseph X. DiMario

6. Single Muscle-Fiber Isolation and Culture for Cellular, Molecular, Pharmacological, and Evolutionary Studies

Judy E. Anderson, Ashley C. Wozniak, and Wataru Mizunoya

7. Somite Unit Chronometry to Analyze Teratogen Phase Specificity in the Paraxial Mesoderm

Sara J. Venters and Charles P. Ordahl

Part II: Non-Mammalian Models of Myogenesis

8. Analysis of Skeletal Muscle Development in Drosophila

Ginny R. Morriss, Anton L. Bryantsev, Maria Chechenova, Elisa M. LaBeau, TyAnna L. Lovato, Kathryn M. Ryan, and Richard M. Cripps

9. Immunocytochemistry to Study Myogenesis in Zebrafish

Nathan C. Bird, Stefanie E. Windner, and Stephen H. Devoto

10. Immunofluorescent Localization of Proteins in C. elegans Muscle

Kristy J. Wilson, Hiroshi Qadota, and Guy M. Benian

Part III: Experimental Models and Analysis of Skeletal Muscle Exercise and Disuse

11. Resistance Loading and Signaling Assays for Oxidative Stress in Rodent Skeletal Muscle

Stephen E. Alway and Robert G. Cutlip

12. Analysis of Skeletal Muscle Hypertrophy in Models of Increased Loading

Sue C. Bodine and Keith Baar

13. Protein Overexpression in Skeletal Muscle Using Plasmid-Based Gene Transfer to Elucidate Mechanisms Controlling Fiber Size

Chia-Ling Wu and Susan C. Kandarian

14. In Vivo Measurement of Muscle Protein Synthesis Rate Using the Flooding Dose Technique

Marta L. Fiorotto, Horacio A. Sosa, Jr., and Teresa A. Davis

Part IV: Generation of Viral Vectors and Transgenic Mice

15. Recombinant Adeno-Associated Viral Vector Production and Purification

Jin-Hong Shin, Yongping Yue, and Dongsheng Duan

16. Generation of Lentiviral Vectors for Use in Skeletal Muscle Research

Christophe Pichavant and Jacques P. Tremblay

17. Generating Tamoxifen-Inducible Cre Alleles to Investigate Myogenesis in Mice

Christoph Lepper and Chen-Ming Fan

Part V: Muscle Profiling

18. Gene Profiling Studies in Skeletal Muscle by Quantitative Real-Time Polymerase Chain Reaction Assay

Shephali Bhatnagar, Siva K. Panguluri, and Ashok Kumar

19. Analysis of Lipid Profiles in Skeletal Muscles

Vassilis Mougios and Anatoli Petridou

20. Proteomic Analysis of Dystrophic Muscle

Caroline Lewis, Philip Doran, and Kay Ohlendieck

Part VI: Experimental Approaches in Calcium Imaging of Skeletal Muscle

21. Detection of Calcium Release via Ryanodine Receptors

Jerry P. Eu and Gerhard Meissner

22. Measurement of Calcium Release Due to Inositol Trisphosphate Receptors in Skeletal Muscle

Mariana Casas, Francisco Altamirano, and Enrique Jaimovich

23. Detection of Calcium Sparks in Intact and Permeabilized Skeletal Muscle Fibers

Noah Weisleder, Jingsong Zhou, and Jianjie Ma

24. Analysis of Calcium Transients in Cardiac Myocytes and Assessment of the Sarcoplasmic Reticulum Ca2+-ATPase Contribution

Anand Mohan Prasad and Giuseppe Inesi

Part VII: Analysis of Gene Promoter Transcriptional Activity

25. Analysis of Muscle Gene Transcription in Cultured Skeletal Muscle Cells

Charis L. Himeda, Phillip W.L. Tai, and Stephen D. Hauschka

26. Analysis of Fiber Type Differences in Reporter Gene Expression of b-Gal Transgenic Muscle

Phillip W.L. Tai, Catherine L. Smith, John C. Angello, and Stephen D. Hauschka

27. Determination of Gene Promoter Activity in Skeletal Muscles In Vivo

Sarah M. Senf and Andrew R. Judge

Part VIII: RNA-Mediated Gene Regulation

28. Determination of MiRNA Targets in Skeletal Muscle Cells

Zhan-Peng Huang, Ramon Espinoza-Lewis, and Da-Zhi Wang

29. shRNA-Mediated Gene Knockdown in Skeletal Muscle

Muriel Golzio, Jean-Michel Escoffre, and Justin Teissie

Part IX: Analysis of Protein-DNA Interactions

30. Detection of NF-kB Activity in Skeletal Muscle Cells by Electrophoretic Mobility Shift Analysis

Jason M. Dahlman and Denis C. Guttridge

31. Isolation of Nuclei from Skeletal Muscle Satellite Cells and Myofibers for Use in Chromatin Immunoprecipitation Assays

Yasuyuki Ohkawa, Chandrashekara Mallappa, Caroline S. Dacwag Vallaster, and Anthony N. Imbalzano

32. An Improved Restriction Enzyme Accessibility Assay (REAA) for Analyzing Changes in Chromatin Structure in Samples of Limited Cell Number

Yasuyuki Ohkawa, Chandrashekara Mallappa, Caroline S. Dacwag Vallaster, and Anthony N. Imbalzano

33. ChIP Enriched in Silico Targets (ChEST), a ChIP on Chip Approach Applied to Analyzing Skeletal Muscle Genes

Guillaume Junion and Krzysztof Jagla