A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes |
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| Authors: | Hieng Chiong Tie Divyanshu Mahajan Bing Chen Li Cheng Antonius M J VanDongen Lei Lu |
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| Institution: | Carnegie Mellon University;aSchool of Biological Sciences, Nanyang Technological University, Singapore 637551;bBioinformatics Institute, Singapore 138671;cSchool of Computing, National University of Singapore, Singapore 117417;dProgram in Neuroscience and Behavioral Disorders, Duke-NUS Graduate Medical School, Singapore 169857 |
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| 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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