Full Description
This volume covers a wide range of approaches utilized to decipher cellular and molecular mechanisms that contribute to successful nerve regeneration leading to functional recovery. Chapters detail a variety of models utilizing both in vivo and in vitro approaches, physical injury models, methods for the isolation/analysis of various macromolecules, live and fixed imaging of regenerating axons, and for quantifying behavioral endpoints enable measurements of regenerative success. 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 methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Axon Regeneration: Methods and Protocols aims to be comprehensive guide for researchers.
Contents
Defining selective neuronal resilience and identifying targets for neuroprotection and axon regeneration using single-cell RNA sequencing: Experimental approaches.- Defining selective neuronal resilience and identifying targets of neuroprotection and axon regeneration using single-cell RNA sequencing - computational approaches.- Retinal Ganglion Cell Axon Fractionation.- Analysis of Immediate Early Gene Expression Levels to Interrogate Changes In Cortical Neuronal Activity Patterns Upon Vision Loss.- Non-coding regulatory RNAs: Isolation and analysis of neuronal circular RNAs and microRNAs.- Transneuronal delivery of cytokines to stimulate mammalian spinal cord regeneration.- Epigenomic Profiling of Dorsal Root Ganglia Upon Regenerative and Non-Regenerative Axonal Injury.- Profiling locally translated mRNAs in regenerating axons.- Analysis of axonal regrowth and dendritic remodeling after optic nerve crush in adult zebrafish.- Assaying OpticNerve Regeneration in Larval Zebrafish.- Surgical Methods in Postmetamorphic Xenopus laevis: Optic Nerve Crush Injury Model.- Generating widespread and scalable retinal lesions in adult zebrafish by intraocular injection of ouabain.- A reproducible spinal cord crush injury in the regeneration permissive axolotl.- Live imaging of axonal dynamics after laser axotomy of peripheral neurons in zebrafish.- Rapid testing of gene function in axonal regeneration after spinal cord injury using larval zebrafish.- Translational Ribosomal Affinity Purification (TRAP) and Bionformatic RNA-Seq Analysis in Postmetamorphic Xenopus laevis.- LCM-seq for Retinal Cell Layer-Specific Responses During Optic Nerve Regeneration.- Temporal-Based Dynamic Profiling of DNA Accessibility With ATAC-Seq in the Regenerating Retina of Zebrafish.- Quantitative Proteomics of Nervous System Regeneration: From Sample Preparation to Functional Data Analyses.- Live Cell Imaging of Dynamic Processes in Adult Zebrafish Retinal Cross-section Cultures.- PCNA staining of retinal cryosections to assess microglial/macrophage proliferation.- Using a Drosophila injury model to investigate RNA repair and splicing in axon regeneration.- Assessing rewiring of the retinal circuitry after inner retinal lesion zebrafish.- Analysis of visual recovery after optic nerve crush in adult zebrafish.
