Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules
Abstract Successful gamete fusion requires species-specific membrane adhesion. However, the interaction of adhesion molecules in gametes is difficult to study in real time through low-throughput microscopic observation. Therefore, we developed a live imaging-based adhesion molecule (LIAM) assay to s...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-13547-w |
| _version_ | 1811251485086842880 |
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| author | Kohdai P. Nakajima Clari Valansi Daisuke Kurihara Narie Sasaki Benjamin Podbilewicz Tetsuya Higashiyama |
| author_facet | Kohdai P. Nakajima Clari Valansi Daisuke Kurihara Narie Sasaki Benjamin Podbilewicz Tetsuya Higashiyama |
| author_sort | Kohdai P. Nakajima |
| collection | DOAJ |
| description | Abstract Successful gamete fusion requires species-specific membrane adhesion. However, the interaction of adhesion molecules in gametes is difficult to study in real time through low-throughput microscopic observation. Therefore, we developed a live imaging-based adhesion molecule (LIAM) assay to study gamete adhesion molecule interactions in cultured cells. First, we modified a fusion assay previously established for fusogens introduced into cultured cells, and confirmed that our live imaging technique could visualise cell–cell fusion in the modified fusion assay. Next, instead of fusogen, we introduced adhesion molecules including a mammalian gamete adhesion molecule pair, IZUMO1 and JUNO, and detected their temporal accumulation at the contact interfaces of adjacent cells. Accumulated IZUMO1 or JUNO was partly translocated to the opposite cells as discrete spots; the mutation in amino acids required for their interaction impaired accumulation and translocation. By using the LIAM assay, we investigated the species specificity of IZUMO1 and JUNO of mouse, human, hamster, and pig in all combinations. IZUMO1 and JUNO accumulation and translocation were observed in conspecific, and some interspecific, combinations, suggesting potentially interchangeable combinations of IZUMO1 and JUNO from different species. |
| first_indexed | 2024-04-12T16:21:09Z |
| format | Article |
| id | doaj.art-97b3d56fdc484231b8060bf4b1c73a2a |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-12T16:21:09Z |
| publishDate | 2022-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-97b3d56fdc484231b8060bf4b1c73a2a2022-12-22T03:25:34ZengNature PortfolioScientific Reports2045-23222022-06-0112111210.1038/s41598-022-13547-wLive imaging-based assay for visualising species-specific interactions in gamete adhesion moleculesKohdai P. Nakajima0Clari Valansi1Daisuke Kurihara2Narie Sasaki3Benjamin Podbilewicz4Tetsuya Higashiyama5Division of Biological Science, Graduate School of Science, Nagoya UniversityDepartment of Biology, Technion-Israel Institute of TechnologyJST, PRESTODivision of Biological Science, Graduate School of Science, Nagoya UniversityDepartment of Biology, Technion-Israel Institute of TechnologyDivision of Biological Science, Graduate School of Science, Nagoya UniversityAbstract Successful gamete fusion requires species-specific membrane adhesion. However, the interaction of adhesion molecules in gametes is difficult to study in real time through low-throughput microscopic observation. Therefore, we developed a live imaging-based adhesion molecule (LIAM) assay to study gamete adhesion molecule interactions in cultured cells. First, we modified a fusion assay previously established for fusogens introduced into cultured cells, and confirmed that our live imaging technique could visualise cell–cell fusion in the modified fusion assay. Next, instead of fusogen, we introduced adhesion molecules including a mammalian gamete adhesion molecule pair, IZUMO1 and JUNO, and detected their temporal accumulation at the contact interfaces of adjacent cells. Accumulated IZUMO1 or JUNO was partly translocated to the opposite cells as discrete spots; the mutation in amino acids required for their interaction impaired accumulation and translocation. By using the LIAM assay, we investigated the species specificity of IZUMO1 and JUNO of mouse, human, hamster, and pig in all combinations. IZUMO1 and JUNO accumulation and translocation were observed in conspecific, and some interspecific, combinations, suggesting potentially interchangeable combinations of IZUMO1 and JUNO from different species.https://doi.org/10.1038/s41598-022-13547-w |
| spellingShingle | Kohdai P. Nakajima Clari Valansi Daisuke Kurihara Narie Sasaki Benjamin Podbilewicz Tetsuya Higashiyama Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules Scientific Reports |
| title | Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules |
| title_full | Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules |
| title_fullStr | Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules |
| title_full_unstemmed | Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules |
| title_short | Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules |
| title_sort | live imaging based assay for visualising species specific interactions in gamete adhesion molecules |
| url | https://doi.org/10.1038/s41598-022-13547-w |
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