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Ultrastructural Study of Golgi Duplication in Trypanosoma brucei
Authors:Jordan T. Yelinek  Cynthia Y. He  Graham Warren
Affiliation:Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA;
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore;
Max F. Perutz Laboratories, Dr Bohr Gasse 9, A-1030 Vienna, Austria
Abstract:Golgi duplication in the protozoan parasite Trypanosoma brucei has been tracked using serial thin section three-dimensional reconstructions of transmission electron micrographs. The old Golgi maintains a constant size (∼0.060 μm3) throughout the cell cycle. A morphologically identifiable new Golgi appears at ∼0.20 of the cell cycle (defined by the size of the nucleus and lasting about 9 h) and grows from ∼0.018 μm3 until it is the same size as the old Golgi (by ∼0.55 of the cell cycle). Morphologically identifiable late Golgi appear at ∼0.58 of the cell cycle, but their volume (∼0.036 μm3) did not change significantly. Cryoimmunoelectron microscopy was used to identify candidates for the earliest new Golgi structures, and these comprised clusters of vesicles containing Golgi reassembly stacking protein (GRASP) near an endoplasmic reticulum exit site. These results, combined with earlier fluorescence data, suggest that the new Golgi begins functioning before cisternal stacks are formed.
Keywords:3D reconstruction    electron microscopy    ER exit site    Golgi    Trypanosoma brucei
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