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This volume represents a valuable and readily reproducible collection of established and emerging techniques for neuronal cell death research. Conveniently divided into four parts, sections cover a series of techniques for the molecular, structural, functional and genomic characterization of dying neurons, a number of protocols that are of primary interest in neuropathology and in experimental neuropathology, a series of gene engineering techniques to obtain and manipulate neuronal stem cells and progenitors, to prepare HSV-1 vectors for the gene therapy, and to CNS transplantation of bone marrow stem cells, and finally, some very interesting protocols for the study of cell death in non-mammalian models. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Neuronal Cell Death: Methods and Protocols seeks to serve a large audience of scientists that are currently active in the field or are willing to enter such an exciting and still expanding area of neurobiology.

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This volume represents a valuable and readily reproducible collection of established and emerging techniques for neuronal cell death research. Conveniently divided into four parts, sections cover a series of techniques for the molecular, structural, functional and genomic characterization of dying neurons, a number of protocols that are of primary interest in neuropathology and in experimental neuropathology, a series of gene engineering techniques to obtain and manipulate neuronal stem cells and progenitors, to prepare HSV-1 vectors for the gene therapy, and to CNS transplantation of bone marrow stem cells, and finally, some very interesting protocols for the study of cell death in non-mammalian models. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

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Authoritative and easily accessible, Neuronal Cell Death: Methods and Protocols seeks to serve a large audience of scientists that are currently active in the field or are willing to enter such an exciting and still expanding area of neurobiology.

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1. Neuronal Cell Death: An Overview of its Different Forms in Central and Peripheral Neurons

Laura Lossi, Claudia Castagna, and Adalberto Merighi
Part I: Molecular, Sturctural, Functional, and Genomic Changes in Dying Neurons2. Cell Volume Regulation Monitored with Combined Epifluorescence and Digital Holographic MicroscopyNicolas Pavillon and Pierre Marquet

3. Flow Cytometric Analysis of DNA Synthesis and Apoptosis in Central Nervous System Using Fresh Cell Nuclei

Noelia Lopez-Sanchez and Jose M. Frade

4. Nuclear Signs of Pre-Neurodegeneration

Fernando C. Baltanas, Jorge Valero, Jose Ramon Alonso, Maria Teresa Berciano, and Miguel Lafarga

5. Multi-Parametric O 2 Imaging in Three-Dimensional Neural Cell Models with the Phosphorescent Probes

Ruslan I. Dmitriev and Dmitri B. Papkovsky

6. Calcium Imaging in Neuron Cell Death

Maria Calvo, Carlos Villalobos, and Lucia Nunez

7. Monitoring Mitochondrial Membranes Permeability in Live Neurons and Mitochondrial Swelling through Electron Microscopy Analysis
Macarena S. Arrazola and Nibaldo C. Inestrosa

8. Real-Time Visualization of Caspase-3 Activation by Fluorescence Resonance Energy Transfer (FRET)

Silvia Alasia, Carolina Cocito, Adalberto Merighi, and Laura Lossi

9. Design and Cloning of Short Hairpin RNAs (shRNAs) into a Lentiviral Silencing Vector to Study the Function of Selected Proteins in Neuronal Apoptosis

Nadia Canu

10. Genomic Analysis Using Affymetrix Standard Microarray GeneChips (169 Format) in Degenerate Murine Retina

Sook Hyun Chung, Weiyong Shen, and Mark Gillies

11. Genomic Analysis of Transcriptional Changes Underlying Neuronal Apoptosis

Sebastiano Cavallaro 

12. High-Throughput Cell Death Assays

Matthew E. Pamenter and Gabriel G. Haddad

PartII: Cell Death in Neuropathology and Experimental Neuoropathology

13. Staining of Dead Neurons by the Golgi Method in Autopsy Material

Stavros J. Baloyannis

14. Image Analysis Algorithms for Immunohistochemical Assessment of Cell Death

Stan Krajewski, Jeffrey Wang, Tashmia Khan, Jonathan Liu, Chia-Hung Sze, and Maryla Krajewska 

15. In Vitro Oxygen-Glucose Deprivation to Study Ischemic Cell Death

Carla I. Tasca, Tharine Dal-Cim, and Helena Cimarosti

16. Laser Microbeam Targeting of Single Nerve Axons in Cell Culture

Nicholas Hyun, Linda Z. Shi, and Michael W. Berns

17. Real-Time Imaging of Retinal Cell Apoptosis by Confocal Scanning Laser Ophthalmoscopy

Eduardo M. Normando, Mohammad H. Dehabadi, Li Guo, Lisa A. Turner, Gaia Pollorsi, and M. Francesca Cordeiro 

18. Targeted Toxicants to Dopaminergic Neuronal Cell Death

Huajun Jin, Arthi Kanthasamy, Dilshan S. Harischandra, Vellareddy Anantharam, Ajay Rana, and Anumantha G. Kanthasamy

Part III: Neural Stem Cells, Progenitors, and Gene Therapy Strategies

19. Stem Cells, Neural Progenitors, and Engineered Stem Cells

Raj R. Rao and Shilpa Iyer

20. Herpes Simplex Virus Type 1 (HSV-1)-Derived Recombinant Vectors for Gene Transfer and Gene Therapy

Peggy Marconi, Cornel Fraefel, and Alberto L. Epstein 

21. Herpes Simplex Virus Type 1 (HSV-1)-Derived Amplicon Vectors for Gene Transfer and Gene Therapy

Cornel Fraefel, Peggy Marconi, and Alberto L. Epstein 

22. Bone Marrow Transplantation for Research and Regenerative Therapies in the Central Nervous System

David Diaz, Jose Ramon Alonso, and Eduardo Weruaga

Part IV: Neuronal Death in Non Mammalian Models

23. Detection of Activated Caspase-8 in Injured Spinal Axons by Using Fluorochrome-Labeled Inhibitors of Caspases (FLICA)

Anton Barreiro-Iglesias and Michael I. Shifman 

24. Generation of Zebrafish Models by CRISPR/Cas9 Genome Editing

Alexander Hruscha and Bettina Schmid

25. In Vivo Assessment of Neuronal Cell Death in Drosophila

Pierre Dourlen

26. Drosophila Model for Studying Phagocytosis Following Neuronal Cell Death 

Boris Shklyar, Flonia Levy-Adam, and Estee Kurant

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