Craniofacial Development, 1st ed. 2022 Methods and Protocols Methods in Molecular Biology Series, Vol. 2403

This volume explores scientific methodologies currently employed to integrate observational developmental biology, tissue explant and cell-based approaches and genetic/molecular technologies to develop a holistic understanding of craniofacial development. Chapters guide readers through the use of disparate models to study formation of the head and face (c. elegans, zebrafish, mouse, alongside human imaging approaches), together with cell culture, tissue explant and in vivo cell imaging and analysis techniques. At the molecular level, chapters include analysing gene expression using in-situ hybridisation and single-cell RNA-Sequencing (scRNA-SEQ), as well as genetic modification techniques such as CRISPR/Cas9-mediated deletion. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. 

1. Using Caenorhabditis elegans as a model for mechanistic insights of craniofacial development

Michael Gruss and Ann K. Corsi

3. Elucidation of Gene Expression Patterns in The Craniofacial Tissues Of Mouse Embryos By Whole-Mount In Situ Hybridization

Joshua B Studdert, Heidi Bildsoe, V Pragathi Masamsetti, and Patrick P L Tam

4. Visualization of the Cartilage and Bone Elements in The Craniofacial Structures by Alcian blue and Alizarin Red Staining

Joshua B Studdert, Heidi Bildsoe, V Pragathi Masamsetti, and Patrick P L Tam

M. E. de Vries and S. Dworkin

6. Methods of Palate Culture in Later Palatogenesis: Elevation, Horizontal Outgrowth, and Fusion

Jack D. Morgan and Jeremy B. A. Green

7. Single cell RNA-seq: Cell Isolation and Data Analysis

Val Yianni and Paul T Sharpe

8. Generating Zebrafish RNA-less Mutant Alleles by Deleting Gene Promoters with CRISPR/Cas9

Priyanka Kumari, Morgan Sturgeon, Gregory Bonde, and Robert A. Cornell

9. Craniofacial Phenomics: Three-Dimensional Assessment of the Size and Shape of Cranial and Dentofacial Structures

Mustafa Mian, Jenny Tan, Robin Yong, Ruth Williams, Agatha Labrinidis, Peter J. Anderson, and Sarbin Ranjitkar

10. Micro-CT-based Bone Microarchitecture Analysis of the Murine Skull

Jenny Tan, Agatha Labrinidis, Ruth Williams, Mustafa Mian, Peter J. Anderson, and Sarbin Ranjitkar

11. Characterization of mammalian in vivo enhancers using mouse transgenesis and CRISPR genome editing

Marco Osterwalder, Stella Tran, Riana D. Hunter, Eman M. Meky, Kianna von Maydell, Anne N. Harrington, Janeth Godoy, Catherine S. Novak, Ingrid Plajzer-Frick, Yiwen Zhu, Jennifer A. Akiyama, Veena Afzal, Evgeny Z. Kvon, Len A. Pennacchio, Diane E. Dickel, and Axel Visel

12. Histological Techniques for Sectioning Bones of the Vertebrate Craniofacial Skeleton

Daniela M. Roth, Lakshmi Puttagunta, and Daniel Graf

13. Engineering Epithelial-Mesenchymal Microtissues to Study Cell-Cell Interactions in Development

Jacob I. Reynolds, Ross A. Vitek, Peter G. Geiger, and Brian P. Johnson

14. Ex Vivo Culture of Human Cranial Suture Cells

Peter J. Anderson

Katherine R. Hixon, Christopher T. Eberlin, Meghana Pendyala, Angela Alarcon de la Lastra, and Scott A. Sell

16. Mandible Explant Assay for The Analysis of Meckel’s Cartilage Development

Sophie Wiszniak and Quenten Schwarz

17. High Resolution Histology for Craniofacial Studies On Zebrafish and Other Teleost Models

18. Live Imaging the Dynamics Of Mammalian Neural Crest Cell Migration

Emma L. Moore and Paul A. Trainor

20. Antenatal Ultrasound Imaging for Analysis of Human Craniosynostosis

Sarah Constantine and Piotr Niznik

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts