2022 Fiscal Year Final Research Report
Development of single-cell analysis of the antimicrobial and bactericidal activities of the antimicrobial peptides and its application
| Project/Area Number |
21K19214
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
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| Research Institution | Shizuoka University |
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Principal Investigator |
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| Project Period (FY) |
2021-07-09 – 2023-03-31
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| Keywords | 抗菌ペプチド / 単一細胞解析 / 抗菌活性 / 殺菌活性 / 抗菌剤 / 細胞膜損傷 / ポア形成 |
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| Outline of Final Research Achievements |
We have developed a novel method to examine the antimicrobial and bactericidal activities of antimicrobial peptides (AMPs) and antibiotics at the single-cell level (i.e., single-cell analysis), and apply this method to examine the mechanism of AMPs’ bactericidal activity. In this method, we monitor the proliferation of single cells on agar in a chamber and measure the distribution of the number of cells in each microcolony. In Method A, we incubate cells in the presence of various concentrations of AMPs for 3 h. AMP concentration dependence of the fraction of microcolonies containing only a single cell, Psingle, provides the minimum inhibitory concentration. In Method B, after the interaction of a cell suspension with an AMP for a specific time, an aliquot is diluted to stop the interaction, and the proliferation of single cells then is monitored after 3-h incubation. The information of Psingle(t) provides the fraction of dead cells after the interaction time t of AMPs with the cells.
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| Free Research Field |
生体膜の生物物理学
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| Academic Significance and Societal Importance of the Research Achievements |
従来の抗菌活性や殺菌活性は、たくさんの細菌細胞の懸濁液を用いる方法（最小発育阻止濃度 (MIC) や最小致死濃度 (MBC) の測定、タイム-キル・アッセイなど）により、たくさんの細胞の応答の平均値の測定から評価されてきた。最近は、1個の細菌細胞とAMPの相互作用の研究もおこなわれるようになり、単一細胞レベルでの抗菌活性や殺菌活性の測定法が必要になってきた。本研究で開発された単一細胞解析法は、単一細胞レベルでのAMPや抗菌剤の抗菌活性および殺菌活性の解析が可能であるので、1個の細胞とAMPの相互作用の研究と組み合わせることにより、AMPの殺菌活性のメカニズムの解明など今後の発展に期待できる
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