Cargando…
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...
Autores principales: | , , , , , |
---|---|
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 |
_version_ | 1782422864176087040 |
---|---|
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. |
format | Online Article Text |
id | pubmed-4803310 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT tiehiengchiong anovelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT mahajandivyanshu anovelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT chenbing anovelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT chengli anovelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT vandongenantoniusmj anovelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT lulei anovelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT tiehiengchiong novelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT mahajandivyanshu novelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT chenbing novelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT chengli novelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT vandongenantoniusmj novelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes AT lulei novelimagingmethodforquantitativegolgilocalizationrevealsdifferentialintragolgitraffickingofsecretorycargoes |