Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools
Small-molecule fluorescent probes are powerful and ubiquitous tools for measuring the concentration and distribution of analytes in living cells. However, accurate characterization of these analytes requires rigorous evaluation of cell-to-cell heterogeneity in fluorescence intensities and intracellu...
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| Format: | Article |
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Springer Nature
2019
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| Online Access: | https://hdl.handle.net/1721.1/121402 |
| _version_ | 1811089950857232384 |
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| author | Han, Yu Goldberg, Jacob Michael Lippard, Stephen J. Palmer, Amy E. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Han, Yu Goldberg, Jacob Michael Lippard, Stephen J. Palmer, Amy E. |
| author_sort | Han, Yu |
| collection | MIT |
| description | Small-molecule fluorescent probes are powerful and ubiquitous tools for measuring the concentration and distribution of analytes in living cells. However, accurate characterization of these analytes requires rigorous evaluation of cell-to-cell heterogeneity in fluorescence intensities and intracellular distribution of probes. In this study, we perform a parallel and systematic comparison of two small-molecule fluorescent vesicular Zn²⁺ probes, FluoZin-3 AM and SpiroZin2, to evaluate each probe for measurement of vesicular Zn²⁺ pools. Our results reveal that SpiroZin2 is a specific lysosomal vesicular Zn²⁺ probe and affords uniform measurement of resting Zn²⁺ levels at the single cell level with proper calibration. In contrast, FluoZin-3 AM produces highly variable fluorescence intensities and non-specifically localizes in the cytosol and multiple vesicular compartments. We further applied SpiroZin2 to lactating mouse mammary epithelial cells and detected a transient increase of lysosomal free Zn²⁺ at 24-hour after lactation hormone treatment, which implies that lysosomes play a role in the regulation of Zn²⁺ homeostasis during lactation. This study demonstrates the need for critical characterization of small-molecule fluorescent probes to define the concentration and localization of analytes in different cell populations, and reveals SpiroZin2 to be capable of reporting diverse perturbations to lysosomal Zn²⁺. |
| first_indexed | 2024-09-23T14:28:08Z |
| format | Article |
| id | mit-1721.1/121402 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T14:28:08Z |
| publishDate | 2019 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | mit-1721.1/1214022022-09-29T09:36:15Z Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools Han, Yu Goldberg, Jacob Michael Lippard, Stephen J. Palmer, Amy E. Massachusetts Institute of Technology. Department of Chemistry Small-molecule fluorescent probes are powerful and ubiquitous tools for measuring the concentration and distribution of analytes in living cells. However, accurate characterization of these analytes requires rigorous evaluation of cell-to-cell heterogeneity in fluorescence intensities and intracellular distribution of probes. In this study, we perform a parallel and systematic comparison of two small-molecule fluorescent vesicular Zn²⁺ probes, FluoZin-3 AM and SpiroZin2, to evaluate each probe for measurement of vesicular Zn²⁺ pools. Our results reveal that SpiroZin2 is a specific lysosomal vesicular Zn²⁺ probe and affords uniform measurement of resting Zn²⁺ levels at the single cell level with proper calibration. In contrast, FluoZin-3 AM produces highly variable fluorescence intensities and non-specifically localizes in the cytosol and multiple vesicular compartments. We further applied SpiroZin2 to lactating mouse mammary epithelial cells and detected a transient increase of lysosomal free Zn²⁺ at 24-hour after lactation hormone treatment, which implies that lysosomes play a role in the regulation of Zn²⁺ homeostasis during lactation. This study demonstrates the need for critical characterization of small-molecule fluorescent probes to define the concentration and localization of analytes in different cell populations, and reveals SpiroZin2 to be capable of reporting diverse perturbations to lysosomal Zn²⁺. National Institutes of Health (U.S.) (Grant GM065519) National Institutes of Health (U.S.) (Grant F32 GM-109516) 2019-06-24T20:43:13Z 2019-06-24T20:43:13Z 2018-09 2018-02 2019-03-25T15:33:07Z Article http://purl.org/eprint/type/JournalArticle 2045-2322 https://hdl.handle.net/1721.1/121402 Han, Yu, Jacob M. Goldberg, Stephen J. Lippard, and Amy E. Palmer. “Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools.” Scientific Reports 8, 1 (October 2018): 15034 © 2018 The Author(s) http://dx.doi.org/10.1038/S41598-018-33102-W Scientific Reports Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Springer Nature Scientific Reports |
| spellingShingle | Han, Yu Goldberg, Jacob Michael Lippard, Stephen J. Palmer, Amy E. Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools |
| title | Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools |
| title_full | Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools |
| title_fullStr | Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools |
| title_full_unstemmed | Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools |
| title_short | Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn²⁺ allows tracking of lysosomal Zn²⁺ pools |
| title_sort | superiority of spirozin2 versus fluozin 3 for monitoring vesicular zn²⁺ allows tracking of lysosomal zn²⁺ pools |
| url | https://hdl.handle.net/1721.1/121402 |
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