Neuronal Network Imaging in Acute Slices Using Ca2+ Sensitive Bioluminescent Reporter

  • Ludovic Tricoire
  • Bertrand Lambolez
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1098)

Abstract

Genetically encoded indicators are valuable tools to study intracellular signaling cascades in real time using fluorescent or bioluminescent imaging techniques. Imaging of Ca2+ indicators is widely used to record transient intracellular Ca2+ increases associated with bioelectrical activity. The natural bioluminescent Ca2+ sensor aequorin has been historically the first Ca2+ indicator used to address biological questions. Aequorin imaging offers several advantages over fluorescent reporters: it is virtually devoid of background signal; it does not require light excitation and interferes little with intracellular processes. Genetically encoded sensors such as aequorin are commonly used in dissociated cultured cells; however it becomes more challenging to express them in differentiated intact specimen such as brain tissue. Here we describe a method to express a GFP-aequorin (GA) fusion protein in pyramidal cells of neocortical acute slices using recombinant Sindbis virus. This technique allows expressing GA in several hundreds of neurons on the same slice and to perform the bioluminescence recording of Ca2+ transients in single neurons or multiple neurons simultaneously.

Key words

Aequorin Calcium imaging Neocortex Sindbis Bioluminescence 
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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Ludovic Tricoire
    • 1
  • Bertrand Lambolez
    • 1
  1. 1.Neurobiologie des processus adaptatifsUMR7102, Université Pierre et Marie CurieParisFrance

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