This volume collects protocols and procedures utilizing cellular, tissue, and whole animal models that can be applied to the investigation of neurotrophic factors and other agents impacting on these systems. It begins with chapters on the culture of neurons and glia from the central and peripheral nervous systems, neuron-glia and glia-glia co-culture models, oligodendrocytes, and cell-based assays for the evaluation of cell vitality. This revised second edition expands to cover methodology encompassing site-specific direct labeling of neurotrophins and their receptors, angiogenesis assays, stem cells, pancreatic beta-cells, axonal transport, synapse biology, dendritic spine analysis, and brain endothelial cells. Subsequent chapters are dedicated to in vivo lesion models of relevance to nervous system pathology, which can be applied to the investigation of neurotrophic factors and peptides, as well as protocols describing nanofiber- and nanoparticle-based methods for brain delivery of neurotrophic agents. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and practical, Neurotrophic Factors: Methods and Protocols, Second Edition is a valuable resource for a wide audience of readers as they explore nervous system function and pathology.