Main description:

This volume provides methods to study bone morphogenetic proteins (BMPs) and BMP modulators in cell culture, in both invertebrate and vertebrate animal models, and for therapeutic applications. Chapters guide the reader in the use of primary and immortalized cell lines, methodologies that exploit transcription and receptor mechanisms, newly developed animal models using nematodes, flies, amphioxus, frogs, zebrafish, and genetically engineered mice, and pre-clinical approaches to understanding BMP function in bone regeneration, heterotopic ossification, and cancers. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Bone Morphogenetic Proteins: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

1. Common Cell Lines Used to Study Bone Morphogenetic Proteins (BMPs)

Jessica Ann Cottrell, Donya Burgess, Sarah Michaels, and Melissa B. Rogers

2. In Vitro and In Vivo Osteogenic Differentiation of Human Adipose-Derived Stromal Cells

Clement D Marshall, Elizabeth A Brett, Alessandra L Moore, Derrick C Wan, and Michael T Longaker

3. Cell-based Gene Therapy System for Delivering BMPs

Austin Dickerson, Eleanor L. Davis, Corinne Sonnet, Alan R. Davis, and Elizabeth A. Olmsted-Davis

4. Generation of Endogenous BMP Transcriptional Reporter Cells through CRISPR/Cas9 Genome Editing

Luke D. Hutchinson, Polyxeni Bozatzi, Thomas Macartney, and Gopal P. Sapkota

5. High-Throughput, Biosensor-based Approach to Examine Bone Morphogenetic Protein (BMP) - Receptor Interactions

Senem Aykul and Erik Martinez-Hackert

6. Mutagenesis and Imaging studies of BMP Signaling Mechanisms in C. elegans

Cathy Savage-Dunn, Ryan J. Gleason, Jun Liu, and Richard W. Padgett

7. Gene Regulation of BMP Ligands in Drosophila

Brian G. Stultz and Deborah A. Hursh

8. Using Amphioxus as a Basal Chordate Model to Study BMP Signaling Pathway

Luok Wen Yong, Iryna Kozmikova, and Jr-Kai Yu

9. Proteolytic Activation of Bmps: Analysis of Cleavage in Xenopus Oocytes and Embryos

Hyung-Seok Kim, Autumn McKnite, and Jan L. Christian

10. Imaging and Quantification of P-Smad1/5 in Zebrafish Blastula and Gastrula Embryos

Joseph Zinski, Francesca Tuazon, Yan Huang, Mary Mullins, and David Umulis

11. An Adult Zebrafish Model of Fibrodysplasia Ossificans Progressiva

Melissa LaBonty and Pamela C. Yelick

12. Generation and Identification of Genetically Modified Mice for BMP Receptors

Jingwen Yang and Yuji Mishina

13. Phenotypic Analyses of Genetically Modified Mice for BMP Receptors

Jingwen Yang and Yuji Mishina

14. Immunofluorescent Visualization of BMP Signaling Activation on Paraffin-embedded Tissue Sections

Maaike Alkema, Marie-Jose Goumans, and Boudewijn P.T. Kruithof

15. Spatial and Quantitative Detection of BMP Activity in Mouse Embryonic Limb Buds

Marcelo Rocha Marques and Jean-Denis Benazet

16. Pharmacologic Strategies for Assaying BMP Signaling Function

Teresa Dinter, Geoffrey A. Bocobo, and Paul B. Yu

17. Bone Morphogenetic Proteins (BMPs) and Bone Regeneration

Deboleena Kanjilal and Jessica Ann Cottrell

18. Heterotopic Ossification in Mouse Models of Fibrodysplasia Ossificans Progressiva

Salin A. Chakkalakal and Eileen M. Shore

19. Double-Humanized Mouse Model to Study Bone Morphogenetic Protein (BMP) Signaling in Tumor Xenografts

Jenna Newman; Rachel NeMoyer, Andrew Zloza, and John Langenfeld