Neurodegeneration Methods and Protocols

1. Front Matter

   Pages i-xi

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2. Introduction to Polymerase Chain Reaction

   • Mark C. Miller, Laura Cunningham

   Pages 1-25

3. Methodological Issues in Polymerase Chain Reaction for RNA

   • Alessandra D. Rasmussen

   Pages 27-35

4. Subtractive Hybridization of cDNA Libraries

   • Gregory S. Kelner, Richard A. Maki

   Pages 37-44

5. The Use of Differential Display RT-PCR for Identifying Altered Gene Expression in Ischemic Brain Injury

   • Xinkang Wang, Giora Z. Feuerstein

   Pages 45-52

6. Analysis of Gene Expression by Multiprobe RNase Protection Assay

   • Anna K. Stalder, Axel Pagenstecher, Carrie L. Kincaid, Iain L. Campbell

   Pages 53-66

7. Molecular Probes for PNS Neurotoxicity, Degeneration, and Regeneration

   • Arrel D. Toews

   Pages 67-87

8. Mobility-Shift DNA-Binding Assay Using Gel Electrophoresis

   • Yang Xiao, Keith Pennypacker

   Pages 89-97

9. Transfection of Mammalian Cells In Vitro: Use in Analysis of Neuronal Damage

   • Melvin L. Billingsley, Theresa A. Thompson

   Pages 99-103

10. Detection of Apoptotic Cells in the Nervous System

    • Terri Kagan, Zahra Zakeri

    Pages 105-123

11. Translocation Assays of Protein Kinase C Activation

    • Ginger G. Wescott, Christina M. Manring, David M. Terrian

    Pages 125-132

12. Astrocyte and Neuron Coculturing Method

    • Michael Aschner, Barbara A. Bennett

    Pages 133-144

13. Viability Assays for Cells In Vitro: The Ethidium/Calcein Assay and the Immunofluorescence Combination Assay

    • Theresa A. Thompson

    Pages 145-155

14. Measurement of the Nitric Oxide Synthase Activity Using the Citrulline Assay

    • Thomas R. Ward, William R. Mundy

    Pages 157-162

15. Measurement of Intraneuronal Free Calcium Using the Fluorescent Probe Technique

    • Prasada Rao S. Kodavanti

    Pages 163-169

16. Measurement of Calcium Buffering by Intracellular Organelles in Brain

    • Prasada Rao S. Kodavanti

    Pages 171-176

17. Receptor-Mediated Release of Inositolphosphates in Brain Slices

    • Pushpa Tandon

    Pages 177-190

18. The Use of Enzyme-Linked Immunosorbent Assays to Quantitate Proteins in Biological Functions

    • Dori R. Germolec

    Pages 191-200

19. Assays for the Analysis of Synaptic Proteins in Neurodegenerative Disorders

    • Eliezer Masliah, Michael Alford

    Pages 201-208

20. Zymographic Method for the Measurement of Gelatinase Activity in Brain Tissue

    • Paul E. Gottschall, J. Wenjun Zhang, Suman Deb

    Pages 209-217

Rapid advances in our understanding of basic cell biological processes and of the molecular mechanisms of cell function and dysfunction have led to an increasing interest in utilizing these approaches in neurobiological research. Efforts in the most rapidly advancing areas are multidisciplinary and the o- come of contributions from many investigators employing a variety of te- niques to address a specific problem. Although a strong basis in fundamental neurobiological concepts is essential for each researcher, the ability to apply new techniques and approaches to the examinination of both cellular and molecular processes requires knowledge of a wide variety of methodologies. The objective of Neurodegeneration Methods and Protocols is to develop an understanding, appreciation, and technical ability in various cellular and molecular techniques for studying many aspects of nervous system cell bi- ogy. The protocols in this volume span a multidisciplinary range of cellular and molecular approaches, and should allow investigators to address research questions directed toward understanding nervous system function, injury, degeneration, and the repair/regenerative process.

". . .the authors are to be commended for the consistency in format and altruism shown contributors to represent the procedures with minimal self-serving recounting of their use of the method."-The Quarterly Review of Biology

• National Institutes of Health, Research Triangle Park

  Jean Harry

• US Environmental Protection Agency, Research Triangle Park

  Hugh A. Tilson

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