Cargando…
A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator
Intracellular vesicles such as lysosomes contain micromolar to millimolar concentrations of Zn(2+), and disturbing lysosomal Zn(2+) homeostasis via lysosomal Zn(2+) release leads to mitochondria damage and consequent lytic cell death. Methods have been developed to image cellular Zn(2+) dynamics. He...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189625/ https://www.ncbi.nlm.nih.gov/pubmed/35707683 http://dx.doi.org/10.1016/j.xpro.2022.101453 |
_version_ | 1784725627418443776 |
---|---|
author | Gu, Mingxue Hu, Meiqin Minckley, Taylor Pinchi, Prateeksunder Xu, Haoxing Qin, Yan Du, Wanlu |
author_facet | Gu, Mingxue Hu, Meiqin Minckley, Taylor Pinchi, Prateeksunder Xu, Haoxing Qin, Yan Du, Wanlu |
author_sort | Gu, Mingxue |
collection | PubMed |
description | Intracellular vesicles such as lysosomes contain micromolar to millimolar concentrations of Zn(2+), and disturbing lysosomal Zn(2+) homeostasis via lysosomal Zn(2+) release leads to mitochondria damage and consequent lytic cell death. Methods have been developed to image cellular Zn(2+) dynamics. Here, we present a protocol using GZnP3, a genetically encoded fluorescent Zn(2+) indicator, to assess lysosomal Zn(2+) release in cultured cells by fluorescence microscopy imaging. For complete details on the use and execution of this protocol, please refer to Du et al. (2021) or Minckley et al. (2019). |
format | Online Article Text |
id | pubmed-9189625 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-91896252022-06-14 A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator Gu, Mingxue Hu, Meiqin Minckley, Taylor Pinchi, Prateeksunder Xu, Haoxing Qin, Yan Du, Wanlu STAR Protoc Protocol Intracellular vesicles such as lysosomes contain micromolar to millimolar concentrations of Zn(2+), and disturbing lysosomal Zn(2+) homeostasis via lysosomal Zn(2+) release leads to mitochondria damage and consequent lytic cell death. Methods have been developed to image cellular Zn(2+) dynamics. Here, we present a protocol using GZnP3, a genetically encoded fluorescent Zn(2+) indicator, to assess lysosomal Zn(2+) release in cultured cells by fluorescence microscopy imaging. For complete details on the use and execution of this protocol, please refer to Du et al. (2021) or Minckley et al. (2019). Elsevier 2022-06-10 /pmc/articles/PMC9189625/ /pubmed/35707683 http://dx.doi.org/10.1016/j.xpro.2022.101453 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Gu, Mingxue Hu, Meiqin Minckley, Taylor Pinchi, Prateeksunder Xu, Haoxing Qin, Yan Du, Wanlu A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator |
title | A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator |
title_full | A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator |
title_fullStr | A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator |
title_full_unstemmed | A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator |
title_short | A protocol to measure lysosomal Zn(2+) release through a genetically encoded Zn(2+) indicator |
title_sort | protocol to measure lysosomal zn(2+) release through a genetically encoded zn(2+) indicator |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189625/ https://www.ncbi.nlm.nih.gov/pubmed/35707683 http://dx.doi.org/10.1016/j.xpro.2022.101453 |
work_keys_str_mv | AT gumingxue aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT humeiqin aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT minckleytaylor aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT pinchiprateeksunder aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT xuhaoxing aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT qinyan aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT duwanlu aprotocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT gumingxue protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT humeiqin protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT minckleytaylor protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT pinchiprateeksunder protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT xuhaoxing protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT qinyan protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator AT duwanlu protocoltomeasurelysosomalzn2releasethroughageneticallyencodedzn2indicator |