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

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 syna...

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Autores principales:	Xu-fei Du, Bing Xu, Yu Zhang, Min-jia Chen, Jiu-lin Du
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Retinotectal Synapses Developmental Synaptogenesis Retinal Ganglion Cells (RGC) Zebrafish Larvae Tectal Neuropil Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6deec56fbc0c450486610c92a3758a64
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spelling	oai:doaj.org-article:6deec56fbc0c450486610c92a3758a642021-12-02T15:08:04ZA transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis10.1038/s41598-018-32409-y2045-2322https://doaj.org/article/6deec56fbc0c450486610c92a3758a642018-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-32409-yhttps://doaj.org/toc/2045-2322Abstract 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. These tools allowed us to investigate molecular events underlying synaptogenesis and found that the microRNA-132 (miR-132) is required for developmental synaptogenesis. Thus, these transgenic zebrafish stable lines provide appropriate tools for studying central synaptogenesis and underlying molecular mechanisms in intact vertebrate brain.Xu-fei DuBing XuYu ZhangMin-jia ChenJiu-lin DuNature PortfolioarticleRetinotectal SynapsesDevelopmental SynaptogenesisRetinal Ganglion Cells (RGC)Zebrafish LarvaeTectal NeuropilMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Retinotectal Synapses Developmental Synaptogenesis Retinal Ganglion Cells (RGC) Zebrafish Larvae Tectal Neuropil Medicine R Science Q
spellingShingle	Retinotectal Synapses Developmental Synaptogenesis Retinal Ganglion Cells (RGC) Zebrafish Larvae Tectal Neuropil Medicine R Science Q Xu-fei Du Bing Xu Yu Zhang Min-jia Chen Jiu-lin Du A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
description	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. These tools allowed us to investigate molecular events underlying synaptogenesis and found that the microRNA-132 (miR-132) is required for developmental synaptogenesis. Thus, these transgenic zebrafish stable lines provide appropriate tools for studying central synaptogenesis and underlying molecular mechanisms in intact vertebrate brain.
format	article
author	Xu-fei Du Bing Xu Yu Zhang Min-jia Chen Jiu-lin Du
author_facet	Xu-fei Du Bing Xu Yu Zhang Min-jia Chen Jiu-lin Du
author_sort	Xu-fei Du
title	A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
title_short	A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
title_full	A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
title_fullStr	A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
title_full_unstemmed	A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
title_sort	transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/6deec56fbc0c450486610c92a3758a64
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A transgenic zebrafish model for in vivo long-term imaging of retinotectal synaptogenesis

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