[ Optical Images ] BDCC-OP-0059_Real-time ex-vivo calcium imaging of downstream neurons upon ATP-induced activation of P2X2-expressing upstream neurons in adult Drosophila brain

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Package Contents
Date of SOP Creation
11.18.2025
Title
BDCC-OP-0059_Real-time ex-vivo calcium imaging of downstream neurons upon ATP-induced activation of P2X2-expressing upstream neurons in adult Drosophila brain
Authors
Gahbien Lee (bien522@ewhain.net)
Yangkyun Oh (yangkyun@ewha.ac.kr)

Department of Life Sciences, Ewha Womans University
Scope and Applicability
This protocol describes the procedure for performing an ex-vivo neuronal circuit activation assay in the adult Drosophila brain using a dual-driver system (GAL4–UAS-P2X2/LexA–LexAop-GCaMP6). Upstream neurons expressing P2X2 are selectively activated by brief ATP perfusion (2.5mM), and the resulting calcium responses in downstream LexA-labeled neurons are monitored in real time using GCaMP6. This method enables precise assessment of functional connectivity between defined neuronal populations under controlled perfusion-chamber conditions.
Keywords
Drosophila melanogaster, adult brain, ex-vivo circuit activation, P2X2, ATP stimulation, GAL4-LexA dual-driver system, GCaMP6, calcium imaging
Instruments
Leica Stellar5 (NFEC-2023-09-250161)