Astrocytes: Methods and Protocols / Edition 1

by Richard Milner

Overview

Ever since their discovery more than 150 years ago, astrocytes have proved to be something of an enigma. Only in the last 10-15 years has it become clear that astrocytes are multifunctional versatile cells that play key roles in a multitude of diverse processes in the CNS, including generation of neural stem cells, synaptogenesis, and regulation of the blood-brain barrier and neurovascular unit. Astrocytes: Methods and Prools provides scientists with a comprehensive guide to many techniques used for astrocyte cell culture, as well as more specialized approaches for studying astrocyte functions, both in vitro and in vivo. Key cellular, molecular and biochemical techniques are used to study the many and varied functions of this fascinating cell. Written in the highly 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 laboratory prools, and key tips on troubleshooting and avoiding known pitfalls.

Product Details

ISBN-13:	9781617794513
Publisher:	Springer-Verlag New York, LLC
Publication date:	12/06/2011
Series:	Methods in Molecular Biology , #814
Edition description:	2012
Pages:	575
Product dimensions:	7.01(w) x 10.00(h) x 0.06(d)

Table of Contents

Part 1. Overview of Astrocyte Biology

1. Astrocytes:  Multi-talented Stars of the Central Nervous System(Ransom-B)

2. Neurogenic Astrocytes and their Glycoconjugates: not just “Glue” Anymore (Steindler-D)

3. Heterogeneity of Astrocytic Form and Function (Goldman-S/ Nedergaard-M)

Part 2. Isolation and Culture of Astrocytes4. Preparation of Mixed Glial Cultures from Postnatal Rat Brain(DeVellis-J)

5. Generation of Primary Astrocyte Cultures Devoid of Contaminating Microglia (Hewett-S

6. Derivation of Microglia-free Astrocyte Cultures from Neural Stem Cells (Milner-R)

7. Isolation and Culture of Spinal Cord Astrocytes (Miller-R

8. Isolation and Culture of Astrocytes from the Retina and Optic Nerve (Lukas-T)

9. Primary Cultures of Astrocytes from Fetal Bovine Brain (Ballarin-C)

10. Isolation and Cultivation of Porcine Astrocytes (Bobilya-D)

11. Isolation and Culture of Human Astrocytes (Sharif-A/ Prevot-V)

12. Cell Culturing of C. elegans Glial Cells for the Assessment of Cytosolic Ca²⁺ Dynamics (Parpura-V)

Part 3. Analysis of Astrocyte Functions In Vitro

13. Astrocyte Polarization and Wound Healing in Culture:Studying Cell Adhesion Molecules (Carbonetto-S)

14. An in Vitro Trauma Model to Study Rodent and Human Astrocyte Reactivity (Wanner-I)

15. Use of Gel Zymography to Examine Matrix Metalloproteinase (gelatinase) Expression in Brain Tissue or in Primary Glial Cultures (del Zoppo-G)

16. ELISA Methodology to Quantify Astrocyte Production of Cytokines/Chemokines In Vitro (Benveniste-T)

17. Induction of Nitric Oxide Synthase-2 Expression and Measurement of Nitric Oxide Production in Enriched Primary Cortical Astrocyte Cultures (Hewett-J)

18. Examining Potassium Channel Function in Astrocytes (Olsen-M)

19. Connexin Based Channels in Astrocytes: How to Study their Properties (Giaume-C)

20. Isotope-based Quantitation of Uptake, Release and Metabolism of Glutamate and Glucose in Cultured Astrocytes (Hertz-L)

Part 4. Co-culture systems to analyse astrocyte interactions with other cell types

21. An in Vitro Model of Reactive Astrogliosis and its Effect on Neuronal Growth (Geller-H)

22. A Neuron-Astrocyte Co-Culture System to Investigate Astrocyte Secreted Factors in Mouse Neuronal Development (Murai-K)

23. Co-cultures of Neurons and Astrocytes as a Model for Examining Hypoxia-induced Neuronal Death (Rempe-D).

24. Use of Astrocyte-Microglial Co-cultures to Examine the Regulatory Influence of Astrocytes on Microglial Activation (Milner-R)

25. Astrocyte-Schwann Cell Co-Culture Systems (Fawcett-J)

26. An in Vitro Assay to Examine Oligodendrocyte Precursor Cell Migration on Astrocytes (Fawcett-J)

27. Investigation of Astrocyte-Oligodendrocyte Interactions in Human Cultures (John-G)

28. An Improved in vitro Blood-Brain Barrier Model: Rat Brain Endothelial Cells Co-Cultured with Astrocytes (Abbott-J)

29. In Vitro Models of the Blood-Brain Barrier (Abbruscato-T)

30. An in Vitro Model of Ischemic Stroke (Abbruscato-T)

31. Pericyte Co-Culture Models to Study Astrocyte, Pericyte, and Endothelial Cell Interactions (Dore-Duffy-P)

Part 5. Analysis of Astrocyte Functions In Vivo

32. Reprogramming of Postnatal Astroglia of the Mouse Neocortex into Functional Synapse-Forming Neurons (Berninger-B)

33. Assessment of Glial Function in the in Vivo Retina (Newman-E)

34. Imaging in Vivo Astrocyte/Endothelial Cell Interactions at the Blood-Brain Barrier (Willis-C)

35. Transgenic Techniques for Cell Ablation or Molecular Deletion to Investigate Functions of Astrocytes and Other GFAP-Expressing Cell Types (Sofroniew-M)

36. Use of Cre-Lox Technology to Analyze Integrin Functions in Astrocytes (McCarty-J)

37. In Vivo Imaging of Ca²⁺ Signaling in Astrocytes Using Two Photon Laser Scanning Fluorescent Microscopy (Ding-S)

Editorial Reviews

Reviewer: Omer Iqbal, MD, FACC, FESC (Loyola University Medical Center)
Description: The advent of sophisticated novel cellular and molecular approaches has prompted rapid strides in the understanding of astrocytes and their important roles in the establishment of neuronal architecture, including regulation of neurogenesis, synaptogenesis, and modulation of synaptic activity. This useful book presents the key cellular, molecular, and biochemical techniques used to learn varied functions of astrocytes.
Purpose: The purpose is to provide a selection of the key techniques for understanding the varied functions of astrocytes.
Audience: It is written for both first-year graduate students and experienced investigators, but other students and clinicians interested in the neurosciences will find this book helpful.
Features: This well-illustrated book is divided into five parts. Part I is an overview of astrocyte biology, part II details the isolation and culture of astrocytes from different species and anatomical locations, part III is an analysis of astrocyte functions in vitro. The 11 chapters in part IV nicely cover co-culture systems used to analyze astrocyte interactions with other cell types, and part V explains the analysis of astrocyte functions in vivo.
Assessment: This is an extremely useful resource for students, experienced investigators, and other clinicians interested in learning the various functions of astrocytes to meet the local metabolic demands of neuronal tissue.

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