T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse
Calcium is an essential cellular messenger that regulates numerous functions in living organisms. Here, we describe development and characterization of ‘Salsa6f’, a fusion of GCaMP6f and tdTomato optimized for cell tracking while monitoring cytosolic Ca2+, and a transgenic Ca2+ reporter mouse with S...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2017-12-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/32417 |
| _version_ | 1818019781626822656 |
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| author | Tobias X Dong Shivashankar Othy Amit Jairaman Jonathan Skupsky Angel Zavala Ian Parker Joseph L Dynes Michael D Cahalan |
| author_facet | Tobias X Dong Shivashankar Othy Amit Jairaman Jonathan Skupsky Angel Zavala Ian Parker Joseph L Dynes Michael D Cahalan |
| author_sort | Tobias X Dong |
| collection | DOAJ |
| description | Calcium is an essential cellular messenger that regulates numerous functions in living organisms. Here, we describe development and characterization of ‘Salsa6f’, a fusion of GCaMP6f and tdTomato optimized for cell tracking while monitoring cytosolic Ca2+, and a transgenic Ca2+ reporter mouse with Salsa6f targeted to the Rosa26 locus for Cre-dependent expression in specific cell types. The development and function of T cells was unaffected in Cd4-Salsa6f mice. We describe Ca2+ signals reported by Salsa6f during T cell receptor activation in naive T cells, helper Th17 T cells and regulatory T cells, and Ca2+ signals mediated in T cells by an activator of mechanosensitive Piezo1 channels. Transgenic expression of Salsa6f enables ratiometric imaging of Ca2+ signals in complex tissue environments found in vivo. Two-photon imaging of migrating T cells in the steady-state lymph node revealed both cell-wide and localized sub-cellular Ca2+ transients (‘sparkles’) as cells migrate. |
| first_indexed | 2024-04-14T07:55:54Z |
| format | Article |
| id | doaj.art-9b0f397a22cd45589cafd282c3004bf4 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:55:54Z |
| publishDate | 2017-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-9b0f397a22cd45589cafd282c3004bf42022-12-22T02:05:03ZengeLife Sciences Publications LtdeLife2050-084X2017-12-01610.7554/eLife.32417T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouseTobias X Dong0https://orcid.org/0000-0001-5500-7099Shivashankar Othy1https://orcid.org/0000-0001-6832-5547Amit Jairaman2Jonathan Skupsky3Angel Zavala4Ian Parker5Joseph L Dynes6Michael D Cahalan7https://orcid.org/0000-0002-4987-2526Department of Physiology and Biophysics, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United States; Department of Medicine, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United States; Department of Neurobiology & Behavior, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United StatesDepartment of Physiology and Biophysics, University of California, Irvine, United States; Institute for Immunology, University of California, Irvine, United StatesCalcium is an essential cellular messenger that regulates numerous functions in living organisms. Here, we describe development and characterization of ‘Salsa6f’, a fusion of GCaMP6f and tdTomato optimized for cell tracking while monitoring cytosolic Ca2+, and a transgenic Ca2+ reporter mouse with Salsa6f targeted to the Rosa26 locus for Cre-dependent expression in specific cell types. The development and function of T cells was unaffected in Cd4-Salsa6f mice. We describe Ca2+ signals reported by Salsa6f during T cell receptor activation in naive T cells, helper Th17 T cells and regulatory T cells, and Ca2+ signals mediated in T cells by an activator of mechanosensitive Piezo1 channels. Transgenic expression of Salsa6f enables ratiometric imaging of Ca2+ signals in complex tissue environments found in vivo. Two-photon imaging of migrating T cells in the steady-state lymph node revealed both cell-wide and localized sub-cellular Ca2+ transients (‘sparkles’) as cells migrate.https://elifesciences.org/articles/32417genetically encoded Ca2+ indicatorCa2+ signalingT cell motilityOrai1two-photon microscopy |
| spellingShingle | Tobias X Dong Shivashankar Othy Amit Jairaman Jonathan Skupsky Angel Zavala Ian Parker Joseph L Dynes Michael D Cahalan T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse eLife genetically encoded Ca2+ indicator Ca2+ signaling T cell motility Orai1 two-photon microscopy |
| title | T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse |
| title_full | T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse |
| title_fullStr | T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse |
| title_full_unstemmed | T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse |
| title_short | T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse |
| title_sort | t cell calcium dynamics visualized in a ratiometric tdtomato gcamp6f transgenic reporter mouse |
| topic | genetically encoded Ca2+ indicator Ca2+ signaling T cell motility Orai1 two-photon microscopy |
| url | https://elifesciences.org/articles/32417 |
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