A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis

Scientific Reports
Xu-Fei DuJiu-Lin Du

Abstract

The retinotectal synapse in larval zebrafish, combined with live time-lapse imaging, provides an advantageous model for study of the development and remodelling of central synapses in vivo. In previous studies, these synapses were labelled by transient expression of fluorescence-tagged synaptic proteins, which resulted in the dramatic variation of labelling patterns in each larva. Here, using GAL4-Upstream Activating Sequence (GAL4-UAS) methodology, we generated stable transgenic lines, which express EGFP-tagged synaptophysin (a presynaptic protein) in retinal ganglion cells (RGCs), to reliably label the pre-synaptic site of retinotectal synapses. This tool avoids the variable labelling of RGCs that occurs in transient transgenic larvae. We obtained several stable transgenic lines that differ consistently in the number of labelled RGCs. Using stable lines that consistently had a single labelled RGC, we could trace synaptogenic dynamics on an individual RGC axonal arbor across different developmental stages. In the stable lines that consistently had multiple labelled RGCs, we could simultaneously monitor both pre- and post-synaptic compartments by combining transient labelling of post-synaptic sites on individual tectal neurons....Continue Reading

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Citations

Jan 29, 2021·Journal of Neuroscience Methods·Shan ZhaoYu Mu
Nov 5, 2020·Diagnostics·Alexander S MachikhinAndrey V Potemkin
Aug 30, 2020·Current Opinion in Neurobiology·Matthew Lovett-Barron
Aug 28, 2021·International Journal of Molecular Sciences·Valentina NaefFilippo M Santorelli

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Methods Mentioned

BETA
dissection
transgenic
PCR

Software Mentioned

Image
ImageJ
PGUSG
GraphPad Prism ( Prism
Pro Plus

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