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Zn2+-Depletion Enhances Lysosome Fission in Cultured Rat Embryonic Cortical Neurons Revealed by a Modified Epifluorescence Microscopic Technique.
Tsao, Hung-Chun; Liao, Yi-Feng; Pratiwi, Feby Wijaya; Mou, Chung-Yuan; Lin, Yi-Jhen; Pan, Chien-Yuan; Chen, Yit-Tsong.
Afiliación
  • Tsao HC; Department of Chemistry, National Taiwan University, Taipei10617, Taiwan.
  • Liao YF; Department of Life Science, National Taiwan University, Taipei10617, Taiwan.
  • Pratiwi FW; Institute of Physics, Academia Sinica, Taipei11529, Taiwan.
  • Mou CY; Department of Chemistry, National Taiwan University, Taipei10617, Taiwan.
  • Lin YJ; Department of Chemistry, National Taiwan University, Taipei10617, Taiwan.
  • Pan CY; Department of Life Science, National Taiwan University, Taipei10617, Taiwan.
  • Chen YT; Department of Life Science, National Taiwan University, Taipei10617, Taiwan.
Microsc Microanal ; 27(2): 420-424, 2021 04.
Article en En | MEDLINE | ID: mdl-33487212
Lysosomes are integration hubs for several signaling pathways, such as autophagy and endocytosis, and also crucial stores of ions, including Zn2+. Lysosomal dysfunction caused by changes in their morphology by fusion and fission processes can result in several pathological disorders. However, the role of Zn2+ in modulating the morphology of lysosomes is unclear. The resolution of conventional epifluorescence microscopy restricts accurate observation of morphological changes of subcellular fluorescence punctum. In this study, we used a modified epifluorescence microscopy to identify the center of a punctum from a series of z-stack images and calculate the morphological changes. We stained primary cultured rat embryonic cortical neurons with FluoZin3, a Zn2+-sensitive fluorescent dye, and Lysotracker, a lysosome-specific marker, to visualize the distribution of Zn2+-enriched vesicles and lysosomes, respectively. Our results revealed that treating neurons with N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine, a cell-permeable Zn2+ chelator, shrank Zn2+-enriched vesicles and lysosomes by up to 25% in an hour. Pretreating the neurons with YM201636, a blocker of lysosome fission, could suppress this shrinkage. These results demonstrate the usefulness of the modified epifluorescence microscopy for investigating the homeostasis of intracellular organelles and related disorders.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Lisosomas / Neuronas Límite: Animals Idioma: En Revista: Microsc Microanal Año: 2021 Tipo del documento: Article País de afiliación: Taiwán Pais de publicación: Reino Unido
Texto completo
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XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Lisosomas / Neuronas Límite: Animals Idioma: En Revista: Microsc Microanal Año: 2021 Tipo del documento: Article País de afiliación: Taiwán Pais de publicación: Reino Unido