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Ca2+ Imaging in Brain Slices Using Bioluminescent Reporters

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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 Aequorin is generally superseded today by fluorescent Ca2+ indicators for imaging applications that require high spatial and temporal resolution. Nonetheless, 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. In this chapter, we describe protocols allowing the expression of a GFP-aequorin fusion protein in acute brain slices and the bioluminescence recording of Ca2+ transients in single neurons, or multiple neurons simultaneously.
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