Main description:

In recent years, the need to develop acceptable alternatives to conventional animal testing for neurotoxicity and developmental neurotoxicity has been increasingly recognized, and much effort is being directed toward the development of alternative models, utilizing mostly mammalian cells in culture but also non-mammalian model systems. In Vitro Neurotoxicology: Methods and Protocols presents a series of cellular, biochemical, and molecular methodological protocols in the area of in vitro neurotoxicology, with an emphasis on mammalian cell culture systems. Opening with a section on methodologies for preparing several cellular systems of variable complexity, amenable for in vitro neurotoxicological studies, the thorough volume continues with coverage of methods to measure cellular death and major mechanisms, methods for assessing mechanisms of nervous system cell toxicity related to impairment of cell signaling, while a final section illustrates additional methods for assessing important nervous system processes such as cell proliferation, neuritogenesis, and synaptogenesis. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, In Vitro Neurotoxicology: Methods and Protocols serves researchers with an interest in assessing or characterizing the potential neurotoxicity of environmental contaminants, drugs, or other chemicals.

Contents:

1. In Vitro Neurotoxicology: An Introduction

Lucio G. Costa, Gennaro Giordano, and Marina Guizzetti

Part I: Model Systems

2. Primary Neurons in Culture and Neuronal Cell Lines for In Vitro Neurotoxicological Studies

Gennaro Giordano and Lucio G. Costa

3. Determination of Metal Interactions with the Chaperone Hspa5 in Human Astrocytoma Cells and Rat Astrocyte Primary Cultures

Evelyn Tiffany-Castiglioni, Yongchang Qian, Rola Barhoumi, and Ying Zheng

4. Microglia Cell Culture: A Primer for the Novice

Anke Witting and Thomas Moeller

5. Neural Stem Cells for Developmental Neurotoxicity Studies

Roshan Tofighi, Michaela Moors, Raj Bose, Wan Norhamidah Wan Ibrahim, and Sandra Ceccatelli

6. Preparation, Maintenance, and Use of Serum-Free Aggregating Brain Cell Cultures

Paul Honegger, Antoinette Defaux, Florianne Monnet-Tschudi, and Marie-Gabrielle Zurich

7. Neurospheres as a Model for Developmental Neurotoxicity Testing

Ellen Fritsche, Kathrin Gassmann, and Timm Schreiber

8. Acute Hippocampal Slice Preparation and Hippocampal Slice Cultures

Pamela J. Lein, Christopher D. Barnhart, and Isaac N. Pessah

9. Glia-Neuron Sandwich Co-Cultures: An In Vitro Approach to Evaluate Cell-to-Cell Communication in Neuroinflammation and Neurotoxicity

Mariaserena Boraso and Barbara Viviani

10. In Vitro Models to Study the Blood Brain Barrier

Hilary Vernon, Katherine Clark, and Joseph P. Bressler

Part II: Cell Death

11. Measurements of Cell Death in Neuronal and Glial Cells

Gennaro Giordano, Sungwoo Hong , Elaine M. Faustman, and Lucio G. Costa

12. Measurements of Neuronal Apoptosis

Gennaro Giordano and Lucio G. Costa

13. Measurement of Isoprostanes as Markers of Oxidative Stress

Dejan Milatovic, Thomas J. Montine, and Michael Aschner

14. Assessment of Glutathione Homeostasis

Gennaro Giordano, Collin C. White, and Lucio G. Costa

15. Assessing Neuronal Bioenergetic Status

Stephanie L.H. Zeiger, Jeannette N. Stankowski, and BethAnn McLaughlin

Part III: Signaling in Neurotoxicity

16. Fluorescent Assessment of Intracellular Calcium in Individual Cells

David P. Cox, Collin C. White, and Terrance J. Kavanagh

17. Homeostatic Regulation of Glutamate Neurotransmission in Primary Neuronal Cultures

Victor Briz and Cristina Sunol

18. Detection of Nitric Oxide Formation in Primary Neural Cells and Tissues

Ronald B. Tjalkens, David L. Carbone, and Guoyao Wu

19. JNK3-Mediated Apoptotic Cell Death in Primary Dopaminergic Neurons

Won-Seok Choi, Heather M Klintworth, and Zhengui Xia

20. Measurement of Proteasomal Dysfunction in Cell Models of Dopaminergic Degeneration

Faneng Sun, Anumantha G. Kanthasamy, and Arthi Kanthasamy

21. Cell Signaling and Neurotoxicity: Protein Kinase C In Vitro and In Vivo

Prasada Rao S. Kodavanti

22. Cell Signaling and Neurotoxicity: 3H-Arachidonic Acid Release (Phospholipase A2) in Cerebellar Granule Neurons

Prasada Rao S. Kodavanti

Part IV: Other End-Points

23. Quantitative Assessment of Neurite Outgrowth in PC12 Cells

Joshua A. Harrill and William R. Mundy

24. Measurements of Astrocyte Proliferation

Marina Guizzetti, Terrance J. Kavanagh, and Lucio G. Costa

25. Quantification of Synaptic Structure Formation in Co-Cultures of Astrocytes and Hippocampal Neurons

Pamela J. Roque, Marina Guizzetti, Gennaro Giordano, and Lucio G. Costa

26. Volume Measurements in Cultured Primary Astrocytes

Michael Aschner

27. Assessment of Cholesterol Homeostasis in Astrocytes and Neurons

Jing Chen, Lucio G. Costa, and Marina Guizzetti

28. Dose-Response or Dose-Effect Curves in In Vitro Experiments and Their Use to Study Combined Effects of Neurotoxicants

Matteo Goldoni and Sara Tagliaferri