Bright and sensitive red voltage indicators for imaging action potentials in brain slices and pancreatic islets
Genetically encoded voltage indicators (GEVIs) allow the direct visualization of cellular membrane potential at the millisecond time scale. Among these, red-emitting GEVIs have been reported to support multichannel recordings and manipulation of cellular activities with reduced autofluorescence background. However, the limited sensitivity and dimness of existing red GEVIs have restricted their applications in neuroscience. Here, we report a pair of red-shifted opsin-based GEVIs, Cepheid1b and Cepheid1s, with improved dynamic range, brightness, and photostability. The improved dynamic range is achieved by a rational design to raise the electrochromic Förster resonance energy transfer efficiency, and the higher brightness and photostability are approached with separately engineered red fluorescent proteins. With Cepheid1 indicators, we recorded complex firings and subthreshold activities of neurons on acute brain slices and observed heterogeneity in the voltage‑calcium coupling on pancreatic islets. Overall, Cepheid1 indicators provide a strong tool to investigate excitable cells in various sophisticated biological systems.
Medienart: |
E-Artikel |
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Erscheinungsjahr: |
2023 |
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Erschienen: |
2023 |
Enthalten in: |
Zur Gesamtaufnahme - volume:9 |
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Enthalten in: |
Science advances - 9(2023), 47 vom: 24. Nov., Seite eadi4208 |
Sprache: |
Englisch |
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Beteiligte Personen: |
Han, Yi [VerfasserIn] |
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Links: |
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Anmerkungen: |
Date Completed 24.11.2023 Date Revised 25.11.2023 published: Print-Electronic Citation Status MEDLINE |
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doi: |
10.1126/sciadv.adi4208 |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
NLM364834463 |
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520 | |a Genetically encoded voltage indicators (GEVIs) allow the direct visualization of cellular membrane potential at the millisecond time scale. Among these, red-emitting GEVIs have been reported to support multichannel recordings and manipulation of cellular activities with reduced autofluorescence background. However, the limited sensitivity and dimness of existing red GEVIs have restricted their applications in neuroscience. Here, we report a pair of red-shifted opsin-based GEVIs, Cepheid1b and Cepheid1s, with improved dynamic range, brightness, and photostability. The improved dynamic range is achieved by a rational design to raise the electrochromic Förster resonance energy transfer efficiency, and the higher brightness and photostability are approached with separately engineered red fluorescent proteins. With Cepheid1 indicators, we recorded complex firings and subthreshold activities of neurons on acute brain slices and observed heterogeneity in the voltage‑calcium coupling on pancreatic islets. Overall, Cepheid1 indicators provide a strong tool to investigate excitable cells in various sophisticated biological systems | ||
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700 | 1 | |a Li, Yuan |e verfasserin |4 aut | |
700 | 1 | |a Chen, Yu |e verfasserin |4 aut | |
700 | 1 | |a Ren, Huixia |e verfasserin |4 aut | |
700 | 1 | |a Ding, Ran |e verfasserin |4 aut | |
700 | 1 | |a Qian, Weiran |e verfasserin |4 aut | |
700 | 1 | |a Ren, Keyuan |e verfasserin |4 aut | |
700 | 1 | |a Xie, Beichen |e verfasserin |4 aut | |
700 | 1 | |a Deng, Mengying |e verfasserin |4 aut | |
700 | 1 | |a Xiao, Yinghan |e verfasserin |4 aut | |
700 | 1 | |a Chu, Jun |e verfasserin |4 aut | |
700 | 1 | |a Zou, Peng |e verfasserin |4 aut | |
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