Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules
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 s...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2024-03-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/81884 |
| _version_ | 1797257833072295936 |
|---|---|
| author | Lei Yang Jingtao Zhang Sen Liu Yanning Zhang Li Wang Xiaotong Wang Shanshan Wang Ke Li Mengping Wei Chen Zhang |
| author_facet | Lei Yang Jingtao Zhang Sen Liu Yanning Zhang Li Wang Xiaotong Wang Shanshan Wang Ke Li Mengping Wei Chen Zhang |
| author_sort | Lei Yang |
| collection | DOAJ |
| description | 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 provide 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 the olfactory bulb, the SYT1-tdTomato signals are highly enriched in the 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. |
| first_indexed | 2024-04-24T22:43:55Z |
| format | Article |
| id | doaj.art-b577a151e9d34dae878d1abc77797951 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-24T22:43:55Z |
| publishDate | 2024-03-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-b577a151e9d34dae878d1abc777979512024-03-18T17:34:16ZengeLife Sciences Publications LtdeLife2050-084X2024-03-011310.7554/eLife.81884Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion moleculesLei Yang0https://orcid.org/0000-0002-2882-5560Jingtao Zhang1Sen Liu2Yanning Zhang3Li Wang4Xiaotong Wang5Shanshan Wang6Ke Li7Mengping Wei8Chen Zhang9https://orcid.org/0000-0002-7940-8054School of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaPeking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, ChinaSchool of Basic Medical Sciences, Beijing Key Laboratory of Neural Regeneration and Repair, Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, China; Chinese Institute for Brain Research, Beijing, ChinaSynapse 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 provide 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 the olfactory bulb, the SYT1-tdTomato signals are highly enriched in the 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.https://elifesciences.org/articles/81884synapse formationSyt1-TDT transgenic miceartificial synapse formation assay |
| spellingShingle | Lei Yang Jingtao Zhang Sen Liu Yanning Zhang Li Wang Xiaotong Wang Shanshan Wang Ke Li Mengping Wei Chen Zhang Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules eLife synapse formation Syt1-TDT transgenic mice artificial synapse formation assay |
| title | Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules |
| title_full | Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules |
| title_fullStr | Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules |
| title_full_unstemmed | Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules |
| title_short | Establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules |
| title_sort | establishment of transgenic fluorescent mice for labeling synapses and screening synaptogenic adhesion molecules |
| topic | synapse formation Syt1-TDT transgenic mice artificial synapse formation assay |
| url | https://elifesciences.org/articles/81884 |
| work_keys_str_mv | AT leiyang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT jingtaozhang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT senliu establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT yanningzhang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT liwang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT xiaotongwang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT shanshanwang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT keli establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT mengpingwei establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules AT chenzhang establishmentoftransgenicfluorescentmiceforlabelingsynapsesandscreeningsynaptogenicadhesionmolecules |