Microcephaly Methods and Protocols

This volume discusses the latest imaging and molecular techniques for studying microcephaly through neural progenitor proliferation, survival, and gene expression. All of the methods covered in this book use the mouse as a model system, and include cellular, metabolic, transcriptomic studies of neural progenitors, MR imaging studies of brain growth in 3-dimensions and developmentally-adapted studies of behavior in neonatal mice. 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 tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Microcephaly: Methods and Protocols is a valuable resource for all researchers who want to expand their knowledge and understanding of this important and developing field.

Dissociation of Cerebellar Granule Neuron Progenitors for Culture, FACS, Transcriptomics, and Molecular Biology.- Maintaining Cerebellar Granule Neuron Progenitors in Cell Culture.- Proliferation Analysis of Cerebellar Granule Neuron Progenitors for Microcephaly Research, Using Immunofluorescent Staining and Flow Cytometry.- Conventional and Spectral Karyotyping of Murine Cerebellar Granule Neuron Progenitors.- Processing of Neonatal Mouse Brains for Immunohistochemical Analysis, As Required for Including Spindle Orientation Analysis and Analysis of DNA Damage and Apoptosis.- Automated Immunofluorescence Staining for Analysis of DNA Damage and Apoptosis in Brain Sections.- Automated Immunofluorescence Staining for Analysis of Mitotic Stages and Division Orientation in Brain Sections.- scRNA-seq for Microcephaly Research [1]: Single-Cell Droplet Encapsulation, mRNA Capture, and cDNA Synthesis.- scRNA-seq for Microcephaly Research [2]: Preparation of Single-Cell cDNA Libraries.- scRNA-seq for Microcephaly Research [3]: Computational Analysis of scRNA-seq Data.- scRNA-seq for Microcephaly Research [4]: Dirichlet Regression for Single Cell Population Differences.- Mouse Brain MRI, Including In Vivo, Ex Vivo, and fcMRI for the Study of Microcephaly.- Whole-Body Mouse Fluxomic Analysis to Detect Metabolic Disruptions Associated with Microcephaly, using ¹³C Isotopes.- Neonatal Behavioral Screen for Mouse Models of Neurodevelopmental Disorders.