Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules

  1. Lei Yang
  2. Jingtao Zhang
  3. Sen Liu
  4. Yanning Zhang
  5. Li Wang
  6. Xiaotong Wang
  7. Shanshan Wang
  8. Ke Li
  9. Mengping Wei  Is a corresponding author
  10. Chen Zhang  Is a corresponding author
  1. School of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, China
  2. Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, China
  3. Chinese Institute for Brain Research, China
6 figures and 1 additional file

Figures

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 …

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 …

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 …

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
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 …

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
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;…

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
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 …

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

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