This project expands the scope of our studies in a cell biology direction. We investigate fusion/docking and sorting/budding in early endosomes (using material mainly from the neuroendocrine cell line PC12). We perform both live-cell and purified organelle assays. The assays are complemented by an investigation of protein domains on endosomes via STED and electron microscopy. Pursuing this project in the direction of investigating endosome-to-Golgi trafficking (as well as Golgi trafficking itself) is envisaged.
The project relies heavily on the use of the photoconversion (or photo-oxidation) technique, in which fluorescent compounds are rendered into electron microscopy markers. Thus, the recycling of synaptic vesicles can be followed in several different preparations, offering an ultrastructural counterpart for the STED studies.
The project was initiated in December 2007, and focuses on the application of super-resolution techniques in living preparations. In line with the direction of the laboratory, the project focuses on the investigation of synaptic vesicle dynamics.
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In this project, which has started in April 2014, we generate a super-resolution image – a nanomap – of presynaptic nerve terminals, which describes the localization of ~100-200 synaptic proteins using STED microscopy. This long-term project consists of a number of shorter-term (6-18 months) sets of experiments, targeting the function and localization of specific protein sets.