Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn2+ allows tracking of lysosomal Zn2+ pools

Abstract 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 i...

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Autores principales:	Yu Han, Jacob M. Goldberg, Stephen J. Lippard, Amy E. Palmer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Lactating Mouse Mammary Epithelial Cells Small-molecule Fluorescent Probes Lactational Hormones Transient Receptor Potential Mucolipin (TRPML1) HC11 Cells Medicine R Science Q
Acceso en línea:	https://doaj.org/article/852635636b5e4591bccbb61e1037cb31
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spelling	oai:doaj.org-article:852635636b5e4591bccbb61e1037cb312021-12-02T15:08:18ZSuperiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn2+ allows tracking of lysosomal Zn2+ pools10.1038/s41598-018-33102-w2045-2322https://doaj.org/article/852635636b5e4591bccbb61e1037cb312018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-33102-whttps://doaj.org/toc/2045-2322Abstract 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+.Yu HanJacob M. GoldbergStephen J. LippardAmy E. PalmerNature PortfolioarticleLactating Mouse Mammary Epithelial CellsSmall-molecule Fluorescent ProbesLactational HormonesTransient Receptor Potential Mucolipin (TRPML1)HC11 CellsMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-15 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Lactating Mouse Mammary Epithelial Cells Small-molecule Fluorescent Probes Lactational Hormones Transient Receptor Potential Mucolipin (TRPML1) HC11 Cells Medicine R Science Q
spellingShingle	Lactating Mouse Mammary Epithelial Cells Small-molecule Fluorescent Probes Lactational Hormones Transient Receptor Potential Mucolipin (TRPML1) HC11 Cells Medicine R Science Q Yu Han Jacob M. Goldberg Stephen J. Lippard Amy E. Palmer Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn2+ allows tracking of lysosomal Zn2+ pools
description	Abstract 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+.
format	article
author	Yu Han Jacob M. Goldberg Stephen J. Lippard Amy E. Palmer
author_facet	Yu Han Jacob M. Goldberg Stephen J. Lippard Amy E. Palmer
author_sort	Yu Han
title	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_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_sort	superiority of spirozin2 versus fluozin-3 for monitoring vesicular zn2+ allows tracking of lysosomal zn2+ pools
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/852635636b5e4591bccbb61e1037cb31
work_keys_str_mv	AT yuhan superiorityofspirozin2versusfluozin3formonitoringvesicularzn2allowstrackingoflysosomalzn2pools AT jacobmgoldberg superiorityofspirozin2versusfluozin3formonitoringvesicularzn2allowstrackingoflysosomalzn2pools AT stephenjlippard superiorityofspirozin2versusfluozin3formonitoringvesicularzn2allowstrackingoflysosomalzn2pools AT amyepalmer superiorityofspirozin2versusfluozin3formonitoringvesicularzn2allowstrackingoflysosomalzn2pools
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Superiority of SpiroZin2 Versus FluoZin-3 for monitoring vesicular Zn2+ allows tracking of lysosomal Zn2+ pools

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