[ Optical Images ] BDCC-OP-0060_Dual-driver (GAL4-P2X2 / LexA-GCaMP6) ex-vivo circuit activation assay for monitoring downstream calcium responses in adult Drosophila enteric neurons

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Package Contents
Date of SOP Creation
11.19.2025
Title
BDCC-OP-0060_Dual-driver (GAL4-P2X2 / LexA-GCaMP6) ex-vivo circuit activation assay for monitoring downstream calcium responses in adult Drosophila enteric neurons
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 enteric neurons (hypocerebral ganglion, HCG) 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 enteric neurons, hypocerebral ganglion (HCG), ex-vivo circuit activation, P2X2, ATP stimulation, GAL4-LexA dual-driver system, GCaMP6, calcium imaging
Instruments
Leica Stellaris5 (NFEC-2023-09-250161)