Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules
- Capital Medical University, China
- Peking University, China
Abstract
Synapse is the fundamental structure for neurons to transmit information between cells. The proper synapse formation is crucial for developing neural circuits and cognitive functions of the brain. The aberrant synapse formation has been proved to cause many neurological disorders, including autism spectrum disorders and intellectual disability. Synaptic cell adhesion molecules (CAMs) are thought to play a major role in achieving mechanistic cell-cell recognition and initiating synapse formation via trans-synaptic interactions. Due to the diversity of synapses in different brain areas, circuits and neurons, although many synaptic CAMs, such as Neurexins (NRXNs), Neuroligins (NLGNs), Synaptic cell adhesion molecules (SynCAMs), Leucine-rich-repeat transmembrane neuronal proteins (LRRTMs) and SLIT and NTRK-like protein (SLITRKs) have been identified as synaptogenic molecules, how these molecules determine specific synapse formation and whether other molecules driving synapse formation remain undiscovered are unclear. Here, to providing a tool for synapse labeling and synaptic CAMs screening by artificial synapse formation (ASF) assay, we generated synaptotagmin-1-tdTomato (Syt1-tdTomato) transgenic mice by inserting the tdTomato-fused synaptotagmin-1 coding sequence into the genome of C57BL/6J mice. In the brain of Syt1-tdTomato transgenic mice, the tdTomato-fused synaptotagmin-1 (SYT1-tdTomato) signals were widely observed in different areas and overlapped with synapsin-1, a widely-used synaptic marker. In olfactory bulb, the SYT1-tdTomato signals are highly enriched in glomerulus. In the cultured hippocampal neurons, the SYT1-tdTomato signals showed colocalization with several synaptic markers. Compared to the wild-type (WT) mouse neurons, cultured hippocampal neurons from Syt1-tdTomato transgenic mice presented normal synaptic neurotransmission. In ASF assays, neurons from Syt1-tdTomato transgenic mice could form synaptic connections with HEK293T cells expressing NLGN2, LRRTM2, and SLITRK2 without immunostaining. Therefore, our work suggested that the Syt1-tdTomato transgenic mice with the ability to label synapses by tdTomato, and it will be a convenient tool for screening synaptogenic molecules.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 1 to 6
Article and author information
Author details
Funding
National Natural Science Foundation of China (81925011)
- Chen Zhang
National Natural Science Foundation of China (92149304)
- Chen Zhang
National Natural Science Foundation of China (32170954)
- Mengping Wei
National Natural Science Foundation of China (32100763)
- Lei Yang
Key-Area Research and Development Program of Guangdong Province (2019B030335001)
- Chen Zhang
Capital Medical University (PXM2021_014226_000026)
- Chen Zhang
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study used C57BL/6J WT mice and Syt1-tdTomato transgenic mice (P0-P56). Animals were housed at room temperature (RT) 20 {plus minus} 2{degree sign}C, with a 12-hour light-dark cycle, air circulating, and unrestricted access to food and water. All animal studies were conducted according to the Guide for the Care and Use of Laboratory Animals (8th edition) and approved by the Animal Experiments and Experimental Animal Welfare Committee of Capital Medical University (Approval ID: AEEI-2019-013).
Copyright
© 2024, Yang et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules
eLife 13:e81884.
https://doi.org/10.7554/eLife.81884
