Description
Nervous system development evolves from the well-orchestrated processes of neural induction, cell proliferation, differentiation, cell migration, survival, and synapse formation. Among these environmental cues, neurotrophic factors are secreted proteins that promote neurite outgrowth, neuronal cell differentiation and survival both in vivo and in vitro. Nerve growth factor (NGF) is the founding and best characterized member of the neurotrophin family of neurotrophic polypeptides. Neurotrophic Factors: Methods and Protocols presents a selection of protocols and procedures which make use of cellular, tissue, and whole animal models which can be applied to the investigation of neurotrophic factors and other agents impacting on these systems. Chapters cover a wide-range of topics such as dealing with the culture of neurons and glia from the central and peripheral nervous systems, neuron-glia co-culture models, and cell-based assays for the evaluation of neuroprotective molecules, as well as assays which can be applied to the study of agents with neuroregenerative potential. Protocols describing viral- and nanoparticle-based delivery methods to neural cells are also presented, following by chapters dealing with organotypic slice culture protocols. Lastly, several 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. Written in the successful Methods in Molecular Biology(TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Neurotrophic Factors: Methods and Protocols seeks to serve both professionals and novices alike with its well-honed methodologies in an effort to further our knowledge of what has been described as the last frontier of science. 1. The Neurotrophin Family of Neurotrophic Factors – An Overview Stephen D. Skaper 2. Neuronal Growth-Promoting and Inhibiting Cues in Neuroprotection and Neurodegeneration Stephen D. Skaper 3. Culture of Rat Cerebellar Granule Neurons and Application to Identify Neuroprotective Agents Laura Facci and Stephen D. Skaper 4. Isolation and Culture of Neural Progenitor Cells from Rat Postnatal Cerebellum Morena Zusso and Patrizia Debetto 5. Culture of Rodent Cortical and Hippocampal Neurons Laura Facci and Stephen D. Skaper 6. Amyloid -Peptide Neurotoxicity Assay Using Cultured Rat Cortical Neurons Laura Facci and Stephen D. Skaper 7. Culture of Neonatal Rodent Microglia, Astrocytes, and Oligodendrocytes from Cortex and Spinal Cord Stephen D. Skaper, Carla Argentini, and Massimo Barbierato 8. Central Nervous System Neuron-Glia Co-Culture Models Stephen D. Skaper and Laura Facci 9. Culture and Characterization of Rat Mesencephalic Dopaminergic Neurons Stephen D. Skaper, Giulia Mercanti, and Laura Facci 10. Preparation of Adult Spinal Cord Motor Neuron Cultures Under Serum-Free Conditions Jose V. Montoya-Gacharna, Jhon Jairo Sutachan, Wai Si Chan, Alexandra Sideris, Thomas J.J. Blanck, and Esperanza Recio-Pinto 11. Rodent Retinal Ganglion Cell Cultures Stephen D. Skaper 12. Culture of Purified Glial Cell Populations from Optic Nerve Stephen D. Skaper 13. Isolation and Culture of Rat Cone Photoreceptor Cells Stephen D. Skaper 14. Rat Retina Pigmented Epithelial Cells Stephen D. Skaper 15. Mammalian Growth Cone Turning Assays Identify Distinct Cell Signalling Mechanisms that Underlie Axon Growth, Guidance and Regeneration Andrew J. Murray, Andrew G. Peace, Steven J. Tucker and Derryck A. Shewan 16. Culture of Dissociated Sensory Neurons from Dorsal Root Ganglia of Postnatal and Adult Rats Davina E. Owen and Julie Egerton 17. Culture and Proliferation of Highly Purified Adult Schwann Cells from Rat, Dog, and Man Kirsten Haastert-Talini 18. Use of PC12 Cells and Rat Superior Cervical Ganglion Sympathetic Neurons as Models for Neuroprotective Assays Relevant to Parkinson’s Disease Cristina Malagelada Grau and Lloyd A. Greene 19. Compartmented Chambers for Studying Neurotrophic Factor Action Stephen D. Skaper 20. Preparation and Culture of Adrenal Chromaffin Cells Natalia Domnguez, Miriam Rodrguez, J. David Machado, and Ricardo Borges 21. Indirect Immunofluorescence Staining of Cultured Neural Cells Massimo Barbierato, Carla Argentini, and Stephen D. Skaper 22. Neurite Outgrowth Assessment Using High Content Analysis Methodology Nicholas M. Radio 23. Dissociated Cell Culture for Testing Effects of Carbon Nanotubes on Neuronal Growth William Lee and Vladimir Parpura 24. High-Resolution Imaging and Evaluation of Spines in Organotypic Hippocampal Slice Cultures Frederik Sndermann, Nataliya Golovyashkina, Roland Brandt, Lidia Bakota, and Christian Tackenberg 25. Imaging Amyloid Precursor Protein In Vivo – An Axonal Transport Assay Toms L. Falzone and Gorazd B. Stokin 26. The Use of Specific AAV Serotypes to Stably Transduce Primary CNS Neuron Cultures Saafan Z. Malik, Margie Maronski, Marc A. Dichter and Deborah J. Watson 27. Preparation and Characterization of Biocompatible Chitosan Nanoparticles for Targeted Brain Delivery of Peptides Secil Caban, Yilmaz Capan, Patrick Couvreur, and Turgay Dalkara 28. [3H]Serotonin Release Assay Using Antigen-Stimulated Rat Peritoneal Mast Cells Stephen D. Skaper and Laura Facci 29. Rat Hippocampal Slice Culture Models for the Evaluation of Neuroprotective Agents Elisabetta Gerace, Elisa Landucci, Tania Scartabelli, Flavio Moroni, and Domenico E. Pellegrini-Giampietro 30. A 6-Hydroxydopamine In Vivo Model of Parkinson’s Disease Giulia Mercanti, Gianfranco Bazzu, and Pietro Giusti 31. Brain Microdialysis in Freely Moving Animals Gianfranco Bazzu, Alice Biosa, Donatella Farina, Ylenia Spissu, Giammario Calia, Sonia Dedola, Gaia Rocchitta, Rossana Migheli, Pier Andrea Serra, and Maria Speranza Desole 32. Evaluating Motor Neuron Death in Neonatal Rats Subjected to Sciatic Nerve Lesion Andre Schwambach Vieira, Alexandre Cesar Santos de Rezende, and Fabio Rogerio 33. Rodent Spinal Cord Injury Model and Application of Neurotrophic Factors for Neuroprotection Hari Shanker Sharma and Aruna Sharma
