Research objectives and content
Through the application of a new electron microcopic procedure, high pressure freezing, we will obtain new detailed information about the structure of polarized MDCK cells. This procedure will give a better represention of the actual situation inside living cells. The combined use of this technique with immuno labelling and tracing procedures, will able us to characterise different types of exocytotic vesicles that pinch of from the trans-Golgi network and that are destined for the basolateral and apical surface of the MDCK cell. With the aid of markers of these vesicles and markers for the endocytotic pathway we will determine whether the exocytotic vesicles intermingle with the endocytotic pathway as they are transported to he surface.
Training content (objective, benefit and expected impact)
The use of high pressure freezing is a relatively new technique. Since it better represents the situation in living cells, the use and demand for this technique will increase over the next years and will be an important tool for ultrastructural investigations. Techniques from different disciplines such as cell biology, molecular biology and biochemistry will be combined in this project, and such a multidisciplinary approach will be a fruitfull base for research.
Links with industry / industrial relevance (22)