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A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes

Cellular functions of the Golgi are determined by the unique distribution of its resident proteins. Currently, electron microscopy is required for the localization of a Golgi protein at the sub-Golgi level. We developed a quantitative sub-Golgi localization method based on centers of fluorescence ma...

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Detalles Bibliográficos
Autores principales: Tie, Hieng Chiong, Mahajan, Divyanshu, Chen, Bing, Cheng, Li, VanDongen, Antonius M. J., Lu, Lei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4803310/
https://www.ncbi.nlm.nih.gov/pubmed/26764092
http://dx.doi.org/10.1091/mbc.E15-09-0664
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author Tie, Hieng Chiong
Mahajan, Divyanshu
Chen, Bing
Cheng, Li
VanDongen, Antonius M. J.
Lu, Lei
author_facet Tie, Hieng Chiong
Mahajan, Divyanshu
Chen, Bing
Cheng, Li
VanDongen, Antonius M. J.
Lu, Lei
author_sort Tie, Hieng Chiong
collection PubMed
description Cellular functions of the Golgi are determined by the unique distribution of its resident proteins. Currently, electron microscopy is required for the localization of a Golgi protein at the sub-Golgi level. We developed a quantitative sub-Golgi localization method based on centers of fluorescence masses of nocodazole-induced Golgi ministacks under conventional optical microscopy. Our method is rapid, convenient, and quantitative, and it yields a practical localization resolution of ∼30 nm. The method was validated by the previous electron microscopy data. We quantitatively studied the intra-Golgi trafficking of synchronized secretory membrane cargoes and directly demonstrated the cisternal progression of cargoes from the cis- to the trans-Golgi. Our data suggest that the constitutive efflux of secretory cargoes could be restricted at the Golgi stack, and the entry of the trans-Golgi network in secretory pathway could be signal dependent.
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spelling pubmed-48033102016-05-16 A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes Tie, Hieng Chiong Mahajan, Divyanshu Chen, Bing Cheng, Li VanDongen, Antonius M. J. Lu, Lei Mol Biol Cell Articles Cellular functions of the Golgi are determined by the unique distribution of its resident proteins. Currently, electron microscopy is required for the localization of a Golgi protein at the sub-Golgi level. We developed a quantitative sub-Golgi localization method based on centers of fluorescence masses of nocodazole-induced Golgi ministacks under conventional optical microscopy. Our method is rapid, convenient, and quantitative, and it yields a practical localization resolution of ∼30 nm. The method was validated by the previous electron microscopy data. We quantitatively studied the intra-Golgi trafficking of synchronized secretory membrane cargoes and directly demonstrated the cisternal progression of cargoes from the cis- to the trans-Golgi. Our data suggest that the constitutive efflux of secretory cargoes could be restricted at the Golgi stack, and the entry of the trans-Golgi network in secretory pathway could be signal dependent. The American Society for Cell Biology 2016-03-01 /pmc/articles/PMC4803310/ /pubmed/26764092 http://dx.doi.org/10.1091/mbc.E15-09-0664 Text en © 2016 Tie et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Tie, Hieng Chiong
Mahajan, Divyanshu
Chen, Bing
Cheng, Li
VanDongen, Antonius M. J.
Lu, Lei
A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes
title A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes
title_full A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes
title_fullStr A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes
title_full_unstemmed A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes
title_short A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes
title_sort novel imaging method for quantitative golgi localization reveals differential intra-golgi trafficking of secretory cargoes
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4803310/
https://www.ncbi.nlm.nih.gov/pubmed/26764092
http://dx.doi.org/10.1091/mbc.E15-09-0664
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