Fluorescent Markers for Neuron and Glia Imaging

Promoters were PCR-amplified from N₂ genomic DNA and then recombined with specific donor vector fragments using the In-Fusion PCR Cloning Kit (TaKaRa Inc.). The following promoter fragments constructed with GFP or mCherry were used in this study: Psra-6::GFP (ASH neuron) and Pvap-1::mCherry (AMsh glia), which were used to identify ASH neurons and AMsh glia, respectively. Psra-6::GCaMP5.0 and Pvap-1::GCaMP5.0 were used for calcium imaging in ASH neurons and AMsh glia, respectively, as previously described (Ding et al., 2015 (link); Duan et al., 2020 (link)).

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Chen D., Cheng H., Liu S., Al-Sheikh U., Fan Y., Duan D., Zou W., Zhu L, & Kang L. (2022). The Voltage-Gated Calcium Channel EGL-19 Acts on Glia to Drive Olfactory Adaptation. Frontiers in Molecular Neuroscience, 15, 907064.

Publication 2022

In-Fusion PCR Cloning Kit

Amsh Calcium Donor Genomic Glia Neurons Pvap Vector

Corresponding Organization : Second Affiliated Hospital of Zhejiang University

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Variable analysis

independent variables

Promoter fragments used: Psra-6, Pvap-1

dependent variables

Expression of GFP in ASH neurons
Expression of mCherry in AMsh glia
Calcium imaging in ASH neurons using Psra-6::GCaMP5.0
Calcium imaging in AMsh glia using Pvap-1::GCaMP5.0

control variables

N2 genomic DNA used as source for promoter fragments
In-Fusion PCR Cloning Kit (TaKaRa Inc.) used for recombination

controls

No positive or negative controls explicitly mentioned

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