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 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.
Duke Scholars
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- trans-Golgi Network
- Viral Envelope Proteins
- Time-Lapse Imaging
- Reproducibility of Results
- Proteins
- Protein Transport
- Nocodazole
- Microscopy, Confocal
- Membrane Glycoproteins
- Humans
Citation
Published In
DOI
EISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- trans-Golgi Network
- Viral Envelope Proteins
- Time-Lapse Imaging
- Reproducibility of Results
- Proteins
- Protein Transport
- Nocodazole
- Microscopy, Confocal
- Membrane Glycoproteins
- Humans