title: A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes.
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.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore 637551.
authors.forename: Hieng Chiong
authors.initials: HC
authors.lastname: Tie
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore 637551.
authors.forename: Divyanshu
authors.initials: D
authors.lastname: Mahajan
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore 637551.
authors.forename: Bing
authors.initials: B
authors.lastname: Chen
authors.affiliation: Bioinformatics Institute, Singapore 138671 School of Computing, National University of Singapore, Singapore 117417.
authors.forename: Li
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Program in Neuroscience and Behavioral Disorders, Duke-NUS Graduate Medical School, Singapore 169857.
authors.forename: Antonius M J
authors.initials: AM
authors.lastname: VanDongen
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore 637551 lulei@ntu.edu.sg.
authors.forename: Lei
authors.initials: L
authors.lastname: Lu
mesh_terms: D002460:Cell Line; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells / Q000187:drug effects; D006801:Humans; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D015739:Nocodazole / Q000494:pharmacology; D021381:Protein Transport; D011506:Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D059008:Time-Lapse Imaging / Q000379:methods*; D014759:Viral Envelope Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D011506:Proteins; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins; D015739:Nocodazole