Molecular characterization of plant endocytosis.

• Other Authors: Lam, Sheung Kwan.
• Format: Others
• Language: English; Chinese
• Published: 2008
• Subjects: Endocytosis; Plant cells and tissues; Plant translocation; Rice > Molecular genetics; Oryza > genetics; Plants > cytology
• Online Access: http://library.cuhk.edu.hk/record=b6074629; http://repository.lib.cuhk.edu.hk/en/item/cuhk-344262

Endocytosis is essential for eukaryotic cells. However, relatively little is known about the endocytic pathway and its molecular machinery in plant cells. In this research, a highly conserved membrane protein called secretory carrier membrane protein 1 (SCAMP) from rice (Oryza sativa) (OsSCAMP1) was employed as a tool to study the plant endocytosis. Toward this goal, I have generated polyclonal antibodies specific to SCAMP and transgenic tobacco BY-2 cell lines expressing yellow fluorescence protein (YFP)-SCAMP fusion. Confocal microscopy study showed that SCAMP localized to both plasma membrane (PM) and motile organelles. Further drug treatment and uptake studies demonstrated that these organelles are early endosomes distinct from Golgi and prevacuolar compartment (PVC), because they colocalized with the endosomal marker FM4-64. Immunogold electron microscopy study with SCAMP antibodies has identified the early endosome (EE) as a vesicular tubular membrane organelle, which resembles the structure of trans-Golgi network (TGN). These results indicate that the secretory and endocytic pathways are merged at the TGN which may serve as the sorting station for both pathways. === Since brefeldin A (BFA) induced both TGN and Golgi to form similar aggregates or BFA compartments in tobacco BY-2 cells, studies were also performed to sort out these BFA-induced compartments. Here I have demonstrated that the BFA-induced compartments derived from Golgi and TGN are physically distinct where the TGN aggregates were usually found to be surrounded by the Golgi aggregates in the same cells in both confocal immunofluorescent and immunogold EM studies. Furthermore, the internalized endosomal marker FM4-64 was found to colocalize with the TGN-derived BFA compartments but separated from the Golgi aggregates, whereas the endocytosis inhibitor tyrphostin A23 prevented TGN but not Golgi from forming BFA compartments. Therefore, the secretory Golgi organelle is functinally distinct from the endocytic TGN/EE in their responses to BFA treatment in plant cells. === The possible roles of SCAMPs in cytokinesis were also investigated. In transgenic tobacco BY-2 cells expressing the TGN/EE marker SCAMP-YFP, SCAMPs were found to be concentrated in the developing cell plate together with the internalized endosomal marker FM4-64 under confocal microscopy and this was further confirmed by immunogold electron microscopy studies with SCAMP antibodies. These results have demonstrated that SCAMPs, TGN and endocytosis are all involved in the cell plate formation during cytokinesis in plant cells. === Lam, Sheung Kwan. === Adviser: Jiang Liwen. === Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3266. === Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. === Includes bibliographical references (leaves 176-191). === Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. === Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. === Abstracts in English and Chinese. === School code: 1307.