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Calcium imaging and BAPTA loading of amygdala astrocytes in mouse brain slices.

STAR protocols (2022-02-25)
Angel Baudon, Etienne Clauss-Creusot, Pascal Darbon, Ryan Patwell, Valery Grinevich, Alexandre Charlet
RÉSUMÉ

Astrocytes are glial cells that exhibit calcium signaling-mediated activity. Here, we present a protocol to monitor and manipulate astrocyte calcium activity from mouse amygdala slices. In the first part of this protocol, we describe the procedure of astrocyte calcium imaging. In the second part, we detail how to disrupt astrocyte calcium activity by patch-clamp-mediated loading of BAPTA. These two approaches are presented separately but they can also be used simultaneously to monitor the effects of disruption on an astrocyte network. For complete details on the use and execution of this protocol, please refer to Wahis et al. (2021).

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HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
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N-Acetyl-L-cysteine, Sigma Grade, ≥99% (TLC), powder
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Guanosine 5′-triphosphate sodium salt hydrate, ≥90% (HPLC)