An efficient autometallography approach to localize lead at ultra-structural levels of cultured cells.

Autor: Song H; Department of Health Service, PLA General Hospital, Beijing 100853, China.; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China., Zheng G; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China., Shen XF; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China., Zhao ZH; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China., Liu Y; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China., Liu Y; Department of Neurology, Xijing hospital, Fourth Military Medical University, Xi'an 710032, China., Liu YY; Institute of Neurosciences, Fourth Military Medical University, Xi'an 710032, China., Kang JJ; Institute of Neurosciences, Fourth Military Medical University, Xi'an 710032, China., Chen JY; Department of Health Service, PLA General Hospital, Beijing 100853, China.; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China., Luo WJ; Department of Occupational and Environmental Health and the Ministry-of-Education's Key Laboratory of Hazard Assessment and Control in Special Operational Environment, School of Public Health, Fourth Military Medical University, Xi'an 710032, China.
Jazyk: angličtina
Zdroj: Biophysics reports [Biophys Rep] 2020 Oct 31; Vol. 6 (5), pp. 211-221.
DOI: 10.1007/s41048-020-00116-9
Abstrakt: Understanding the precise intracellular localization of lead (Pb) is a key in deciphering processes in Pb-induced toxicology. However, it is a great challenge to trace Pb in vitro , especially in cultured cells. We aimed to find an innovative and efficient approach to investigate distribution of Pb in cells and to validate it through determining the subcellular Pb content. We identified its ultra-structural distribution with autometallography under electron microscopy in a choroidal epithelial Z310 cell line. Electron microscopy in combination with energy-dispersive X-ray spectroscope (EDS) was employed to provide further evidence of Pb location. In addition, Pb content was determined in the cytosol, membrane/organelle, nucleus and cytoskeleton fractions with atomic absorption spectroscopy. Pb was found predominantly inside the nuclear membranes and some was distributed in the cytoplasm under low-concentration exposure. Nuclear existence of Pb was verified by EDS under electron microscopy. Once standardized for protein content, Pb percentage in the nucleus fraction reached the highest level (76%). Our results indicate that Pb is accumulated mainly in the nucleus of choroid plexus. This method is sensitive and precise in providing optimal means to study the ultra-structural localization of Pb for in vitro models. In addition, it offers the possibility of localization of other metals in cultured cells. Some procedures may also be adopted to detect target proteins via immunoreactions.
Competing Interests: Han Song, Gang Zheng, Xue-Feng Shen, Zai-Hua Zhao, Yang Liu, Yang Liu, Ying-Ying Liu, Jun-Jun Kang, Jing-Yuan Chen and Wen-Jing Luo declare that they have no conflict of interest.
(Copyright 2020 Biophysics Reports.)
Databáze: MEDLINE
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