Neuronal Morphogenesis Methods and Protocols

This volume details molecular mechanisms that are involved in neuronal morphogenesis, and methods for analyzing these mechanisms. Chapters detail analyzing neuronal morphology in vitro and in vivo using a variety of model systems, including rodents and human induced pluripotent stem cells (hiPSCs). 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, Neuronal Morphogenesis: Methods and Protocols aims to be  suitable for a wide range of readers interested in the mechanisms of neuronal morphology and related neurological disorders.

Culturing primary cortical neurons for live-imaging.- Electron Microscopy of Neurons on Biomimetic Substrates.- Analysis of Neurite and Spine Formation in Neurons In Vitro.- Quantifying Dendritic Arbors In Vitro and In Vivo in Rodent Models.- Visualizing and Measuring Dendrite Arborization in Drosophila Somatosensory Neurons.- Analyses of Neurite and Spine Formation by In Utero Electroporation in Mice.- Analysis of Neuronal Morphology by Two-Photon Microscopy.- In-Ovo Electroporation of Embryonic Chicken Spinal Cord in combination with Ex-Vivo Slice Culture for Live Imaging of Vertebrate Neuronal Differentiation.- In Vitro and In Vivo Analysis of Neuronal Polarity.- Proteomic Identification and Label-Free Quantification of Proteins Implicated in Neurite and Spine Formation.- 3D reconstruction of the mitochondrial network within the neuronal soma from SBF-SEM volume data.- DNeuroMAT: A Deep-Learning Based Neuron Morphology Analysis Toolbox.- Ex Vivo Live Imaging in Brain Slices for Analysis of Neurite Formation in Combination with In Utero Electroporation.- Imaging of Structural Plasticity of Dendritic Spines with Two-Photon Microscopy.- Visualization and Quantification of Organelle Axonal Transport in Cultured Neurons.- Visualization of microtubule polymerization during axon outgrowth using fluorescently labelled microtubule plus-end tracking proteins.- Interrogating the molecular clutch in neuronal growth cones: measuring traction forces, F-actin retrograde flow and point contact demographics.- Measuring Retrograde Actin Flow in Neuronal Growth Cones.- Morphological Analysis of Neurons and Glia using Mosaic Analysis with Double Markers.- Isolation and Culture of Adult Rat Dorsal Root Ganglion Neurons for the In Vitro Analysis of Peripheral Nerve Degeneration and Regeneration.- A live imaging assay for regenerating peripheral neuron.- Nerve Regeneration Using Compartmentalized system in CIPN model.- Live imaging analysis of axonal regeneration in human iPSC-derived motor neurons using a microfluidic system.- In Vivo Neural Tract Tracing Procedures: Unravelling Neural Hodology Using Fluorescent and non-fluorescent neural tract tracers, Optogenetic Approach, and Diffusion Tensor Neurotractography Protocols.