A protocol to measure lysosomal Zn2+ release through a genetically encoded Zn2+ indicator

Autor: Mingxue Gu, Meiqin Hu, Taylor Minckley, Prateeksunder Pinchi, Haoxing Xu, Yan Qin, Wanlu Du
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: STAR Protocols, Vol 3, Iss 2, Pp 101453- (2022)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2022.101453
Popis: Summary: Intracellular vesicles such as lysosomes contain micromolar to millimolar concentrations of Zn2+, and disturbing lysosomal Zn2+ homeostasis via lysosomal Zn2+ release leads to mitochondria damage and consequent lytic cell death. Methods have been developed to image cellular Zn2+ dynamics. Here, we present a protocol using GZnP3, a genetically encoded fluorescent Zn2+ indicator, to assess lysosomal Zn2+ 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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