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Multiphoton Ca2+ Imaging of Astrocytes with Genetically Encoded Indicators Delivered by a Viral Approach

  • Rune Enger
  • Rolf Sprengel
  • Erlend A. NagelhusEmail author
  • Wannan Tang
Protocol
Part of the Neuromethods book series (NM, volume 148)

Abstract

Decades of research have unraveled the complex functioning of neurons in the central nervous system. Our knowledge of the second main player of the brain – the non-excitable glial cells – clearly lags behind that of neurons. Pioneering work in the 1990s provided evidence that star-shaped glial cells – astrocytes – sense and modulate neuronal activity by intracellular Ca2+ signals. However, the precise roles of astrocytic Ca2+ signaling in brain physiology and pathophysiology are still highly controversial, largely due to technical limitations of previous Ca2+ imaging tools. With recent innovations in laser microscopy and engineering of molecular probes, the field of glioscience is undergoing a revolution. This chapter describes the application of multiphoton microscopy and genetically encoded fluorescent Ca2+ indicators to reveal astrocytic Ca2+ signals in acute brain slices and in vivo, both in anesthetized and in awake behaving animals.

Keywords

Astrocytes Ca2+ signaling Genetically encoded calcium indicator GECI GCaMP6 Glia rAAV gene transduction Two-photon microscopy 

Notes

Acknowledgments

This work was supported by grants from South and Eastern Norway Regional Health Authority (2016070); the Research Council of Norway (grants 240476, 249988, and 262552); the European Union’s Seventh Framework Programme for research, technological development, and demonstration under grant agreement no. 601055; and the Letten Foundation.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Rune Enger
    • 1
    • 2
  • Rolf Sprengel
    • 3
  • Erlend A. Nagelhus
    • 1
    • 2
    Email author
  • Wannan Tang
    • 1
    • 2
  1. 1.Oslo University Hospital, Department of NeurologyOsloNorway
  2. 2.GliaLab and Letten Centre, Division of Physiology, Department of Molecular Medicine, Institute of Basic Medical Sciences, University of OsloOsloNorway
  3. 3.Research Group of the Max Planck Institute for Medical Research, Institute for Anatomy and Cell Biology, Heidelberg UniversityHeidelbergGermany

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