Skeletal Muscle Regeneration in the Mouse

This volume focuses on the cell biology and physiology of skeletal muscle regeneration. This Book is a collection of classic and cutting edge protocols optimized for mice, but in most cases adaptable to rat or other mammalian models, that will allow an investigator to develop and implement a research study on skeletal muscle regeneration. Chapters address the three major areas of study: provoking regeneration by inducing damage to muscle, analyzing the progenitor cells of skeletal muscle, and quantifying overall muscle function. Subjects discussed include: inducing skeletal muscle injury by eccentric contraction; volumetric muscle loss; single myofiber isolation and culture; satellite cell transplantation; muscle clearing for whole mount immunostaining; luciferase tracking of muscle stem cells; mitochondrial and mitophagy flux analysis; in vivo assessment of muscle contractility; force measurements on single isolated myofibers; and analysis of aerobic respiration in intact skeletal muscle tissue by microplate respirometry. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to each respective topic, 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 practical, Skeletal Muscle Regeneration in the Mouse: Methods and Protocols is an essential laboratory reference for research in skeletal muscle growth, damage, repair, degeneration, and regenerative therapy in the mouse model system.

Includes cutting-edge methods and protocols Provides step-by-step details essential for reproducible results Contains key notes and implementation advice from the experts

Autorentext
Michael Kyba, Ph.D. Associate Professor and Lillehei Endowed Scholar, Lillehei Heart Institute NHH4.126, University of Minnesota, SE Minneapolis, MN 55455, USA

Inhalt

Eccentric Contraction-Induced Muscle Injury: Reproducible, Quantitative, Physiological Models to Impair Skeletal Muscle's Capacity to Generate Force.- Volumetric Muscle Loss.- Freeze Injury of the Tibialis Anterior Muscle.- Synergist Ablation as a Rodent Model to Study Satellite Cell Dynamics in Adult Skeletal Muscle.- Inducing and Evaluating Skeletal Muscle Injury by Notexin and Barium Chloride.- Cardiotoxin Induced Injury and Skeletal Muscle Regeneration.- Fibrosis-Inducing Strategies in Regenerating Dystrophic and Normal Skeletal Muscle.- Isolation, Cryosection, and Immunostaining of Skeletal Muscle.- Isolation of Mouse Periocular Tissue for Histological and Immunostaining Analyses of the Extraocular Muscles and their Satellite Cells.- Skeletal Muscle Tissue Clearing for LacZ and Fluorescent Reporters, and Immunofluorescence Staining.- Isolation, Culture, Functional Assays and Immunofluorescence of Myofiber-Associated Satellite Cells.- Transplantation-Based Quantitative Assays for Satellite Cell Self-Renewal and Differentiation.- Non-Invasive Tracking of Quiescent and Activated MuSC Engraftment Dynamics In Vivo.- Myogenic Progenitors from Mouse Pluripotent Stem Cells for Muscle Regeneration.- Assaying Human Myogenic Progenitor Cell Activity by Reconstitution of Muscle Fibers and Satellite Cells in Immunodeficient Mice.- Methods for Mitochondria and Mitophagy Flux Analyses in Stem Cells of Resting and Regenerating Skeletal Muscle.- Identification, Isolation, and Characterization of Mesenchymal Progenitors in Mouse and Human Skeletal Muscle.- FACS Fractionation and Differentiation of Skeletal-Muscle Resident Multipotent Tie2+ Progenitors.- In Vitro Assays to Determine Skeletal Muscle Physiologic Function.- In Vivo Assessment of Muscle Contractility in Animal Studies.- Functional Measurement of Respiratory Muscle Motor Behaviors Using Transdiaphragmatic Pressure.- Assessment of the Contractile Properties of Permeabilized Skeletal Muscle Fibers.- Analysis of Aerobic Respiration in Intact Skeletal Muscle Tissue by Microplate-Based Respirometry.