Ultrastructural study of Golgi duplication in Trypanosoma brucei

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 (approximately 0.060 microm(3)) throughout the cell cycle. A morphologically identifiable new Golgi appears at approximately 0.20 of the cell cycle (defined by the size of the nucleus and lasting about 9 h) and grows from approximately 0.018 microm(3) until it is the same size as the old Golgi (by approximately 0.55 of the cell cycle). Morphologically identifiable late Golgi appear at approximately 0.58 of the cell cycle, but their volume ( approximately 0.036 microm(3)) 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.