Microglia: Methods and Protocols

ISBN-13: 9781493960514 / Angielski / Miękka / 2016 / 350 str.

Bertrand Joseph; Jose Luis Venero

Opis

Key discoveries concerning the different biological functions of microglia in health and disease have attracted scientists from various fields. In Microglia: Methods and Protocols, expert researchers in the field detail methods for selection of the key cellular, molecular and biochemical techniques that are used in studying the many and varied functions of this fascinating cell. These methods and techniques include microglia cell culture for studying microglia activation and functions, as well as their interaction with other cell types both in vitro and in vivo. 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 protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Microglia: Methods and Protocols is a useful resource for cell biologists, molecular biologists, immunologists, oncologist and neuroscientists.

Spis treści

Part I: Overview of Microglia Biology

1. A Brief Overview of Multi-Talented Microglia

Bertrand Joseph and Jose Luis Venero

 

Part II: Isolation and Culture of Microglia

2. Cell Culturing of Human and Murine Microglia Cell Lines

Johanna Rodhe

 3. Microglia Isolation from Adult Mouse Brain

Jae-Kyung Lee, Malú G Tansey

 

4. Preparation of Primary Microglia Cultures from Postnatal Mouse and Rat Brains

Tomas Deierborg

 

5. Isolation of Murine Postnatal Brain Microglia for Phenotypic Characterization Using Magnetic Cell Separation Technology

Ashley S. Harms and Malú G. Tansey

 

6. Isolation and Culture of Adult Human Microglia within Mixed Glial Cultures for Functional Experimentation and High Content Analysis

Amy M. Smith, Hannah M. Gibbons, Claire Lill, Richard L. M. Faull and Mike Dragunow

 

Part III: Depletion and Transduction of Microglia

7. Depletion of Microglia from Primary Cellular Cultures

Lorena Pont-Lezica, Sabrina Colasse and Alain Bessis

 

8. Lentiviral Transduction of Cultured Microglia

Takahiro Masuda, Makoto Tsuda, Hidetoshi Tozaki-Saitoh and Kazuhide Inoue

 

Part IV: Analysis of Microglial Cytokine Production

9. Microglial Activation: Measurement of Cytokines by Flow Cytometry

Deepak Kumar Kaushik and AnirbanBasu

 

10. In situ Hybridization of Cytokine mRNA using Alkaline Phosphatase Labelled Oligodeoxynucleotide Probes

Bettina Clausen, Christina Fenger and Bente Finsen

 

11. Use of Meso-scale Discovery^TM  to Examine Cytokinecontentin Microglia Cell Supernatant

Miguel A. Burguillos

 

 

Part V: Analysis of Microglia Activation

12. Analysis of Microglial Production of Reactive Oxygen and Nitrogen Species

UrteNeniskyte and Guy C. Brown

 

13. Quantification of Active Caspase-3 and Active Caspase-8 in Microglia Cells

Edel Kavanagh

 

14. Quantification of Microglial Phagocytosis by a Flow Cytometer-based Assay

Refik Pul, Chittappen Kandiyil Prajeeth, Martin Stangel

 

15. Quantification of Microglial Proliferation and Apoptosis by Flow Cytometry

Alicia A. Babcock, Martin Wirenfeldt, and Bente Finsen

 

16. Fluorescence Imaging of Intracellular Ca²⁺, Na⁺ and H⁺ in Cultured Microglia

Tom Schilling, and Claudia Eder

 

17. Patch Clamp Protocols to Study Ion Channel Activity in Microglia

Tom Schilling, and Claudia Eder

  Part VI: Analysis of Microglia Polarization18. Studying M1 and M2 States in Adult Microglia

Sadanand M. Gaikwad and Michael T. Heneka

 

19. Isolating, Culturing, and Polarizing Primary Human Adult and Fetal Microglia

Bryce A. Durafourt, Craig S. Moore, Manon Blain, and Jack P. Antel

 

 

Part VII: Co-culture Systems to Analysis Microglia Interactions with Other Cell Types

20. Understanding Microglia-neuron Cross-talk: Relevance of the Microglia-neuron Co-cultures

Fernando G Correa, Miriam Hernangómez, and Carmen Guaza

 

21. Preparation of Rodent Primary Cultures for Neuron-Glia, Mixed Glia, Enriched Microglia, and Reconstituted Cultures with Microglia

Shih-Heng Chen, Esteban A. Oyarzabal and Jau-Shyong Hong

 

 

Part VIII: Analysis of Microglia Functions in vivo

22. Microglia Detection by Enzymatic Histochemistry

Beatriz Almolda, Berta González and Bernardo Castellano

 

23. Tomato Lectin Histochemistry for Microglial Visulization

Nàdia Villacampa, Beatriz Almolda, Berta González and Bernardo Castellano

 

24. Immunohistochemical Detection of Microglia

Rocío M. de Pablos, Ana M. Espinosa-Oliva, and Antonio J. Herrera

 

25. Intrathecal Infusion of Microglia Cells

Takahiro Masuda, Makoto Tsuda, Hidetoshi Tozaki-Saitoh and Kazuhide Inoue

 

26. Intracranial Injection of LPS in Rat as Animal Model of Neuroinflammation

Ana M. Espinosa-Oliva, Rocío M. de Pablos, and  Antonio J. Herrera

 

27. Analyses of Microglia Effector Function Using CX3CR1-GFP Knock In Mice

Jenny A. Garcia, Sandra M. Cardona and Astrid E. Cardona

 

28. In vivo Two-photon Microscopy of Microglia

Satoru Kondo and Shigeo Okabe

 

29. Use of Confocal Microscopy in the Study of Microglia in a Brain Metastasis Model

Manuel Sarmiento