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

Molecular Biology of the Cell
Hieng Chiong TieLei Lu

Abstract

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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Citations

Oct 18, 2016·Alcoholism, Clinical and Experimental Research·Carol A CaseyArmen Petrosyan
Dec 12, 2019·The Journal of Cell Biology·Walaa AlshafiePeter S McPherson
Aug 28, 2019·International Journal of Molecular Sciences·Laurent M PaardekooperGeert van den Bogaart
Mar 13, 2019·The Journal of Cell Biology·Jason C CaslerBenjamin S Glick
Aug 27, 2019·Frontiers in Cell and Developmental Biology·Areti Pantazopoulou, Benjamin S Glick
Jan 18, 2020·Nature Communications·Kerrie L MariePravin J Mishra
Aug 23, 2020·The Journal of Biological Chemistry·Xiuping SunLei Lu
Oct 29, 2020·Molecular Biology of the Cell·Jason C CaslerBenjamin S Glick
Mar 12, 2021·Current Opinion in Cell Biology·Bulat R RamazanovJulia von Blume
May 27, 2021·Archives of Biochemistry and Biophysics·Pablo Lujan, Felix Campelo
May 26, 2021·Current Biology : CB·Rajarshi ChakrabartiHenry N Higgs
Sep 18, 2021·Journal of Cell Science·Xiuping SunLei Lu
Nov 6, 2021·The Journal of Cell Biology·Jason C CaslerBenjamin S Glick

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Datasets Mentioned

BETA
GM130

Methods Mentioned

BETA
light microscopy
fluorescence microscopy
superresolution microscopy
PCR
PCRs
transfection

Software Mentioned

Zen
ImageJ
OriginPro8
GLIM
MetaMorph
MATLAB

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