Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules
- School of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, China
- Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, China
- Chinese Institute for Brain Research, China
Figures
Figure 1
Construction of tdTomato-fused synaptotagmin-1 (Syt1-tdTomato) expression vector and transgenic fluorescent mice.
(A) Schematic diagram of expression vector for generating Syt1-tdTomato transgenic mice. (B) The expression vector was digested by restriction enzymes for confirmation purposes. The size of enzyme …
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Figure 1—source data 1
The original gel of panels B, C and D.
- https://cdn.elifesciences.org/articles/81884/elife-81884-fig1-data1-v2.zip
Figure 2
tdTomato-fused synaptotagmin-1 (SYT1-tdTomato) is expressed in different brain regions of transgenic mice.
(A) Representative fluorescence images of the sagittal plane of the brains in Syt1-tdTomato transgenic mice and wild-type (WT) mice (8 weeks). The SYT1-tdTomato signals were shown in channel …
Figure 3
tdTomato-fused synaptotagmin-1 (SYT1-tdTomato) is colocalized with synaptic markers in situ.
(A) The images of SYT1-tdTomato (tdTomato) signals and synapsin1 (SYN1) signals in hippocampus and cerebellum. (B) Magnified images of SYT1-tdTomato and SYN1 in granular layer of cerebellum and the …
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Figure 3—source data 1
Numerical data corresponding to the graph in panels B and C.
- https://cdn.elifesciences.org/articles/81884/elife-81884-fig3-data1-v2.zip
Figure 4
tdTomato-fused synaptotagmin-1 (SYT1-tdTomato) signals localize to the synaptic site in cultured neurons.
(A–B) Representative images of hippocampal neurons from Syt1-tdTomato transgenic mice stained with antibodies against SYN1, VGLUT1, GAD65, PSD95, Homer-1, and Gephyrin at DIV 14. (C) Summary graphs …
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Figure 4—source data 1
Numerical data corresponding to the graph in panel C.
- https://cdn.elifesciences.org/articles/81884/elife-81884-fig4-data1-v2.zip
Figure 5
No significant changes in electrophysiological properties were observed in neurons of tdTomato-fused synaptotagmin-1 (Syt1-tdTomato) transgenic mice.
(A–C) The spontaneous action potentials were recorded in hippocampal neurons of wild-type (WT) mice and Syt1-tdTomato transgenic mice (TG) at DIV 13–15. (A) Representative traces of action potential;…
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Figure 5—source data 1
Numerical data corresponding to the graph in panel C, F, I.
- https://cdn.elifesciences.org/articles/81884/elife-81884-fig5-data1-v2.zip
Figure 6
tdTomato-fused synaptotagmin-1 (Syt1-tdTomato) transgenic mice hippocampal neurons can be used in artificial synapse formation experiments to screen for synaptogenic molecules.
(A) Representative fluorescent images of co-cultured neurons from Syt1-tdTomato transgenic mice with HEK293T cells transfected with plasmid encoding NLGN2 and GFP, which are also stained with …
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Figure 6—source data 1
Numerical data corresponding to the graph in panel C.
- https://cdn.elifesciences.org/articles/81884/elife-81884-fig6-data1-v2.zip
