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Dr Shevchuk and his team have established a method that allows them to study the effects of mutations in the dynamin 2 gene that are linked to centronuclear myopathy (CNM) and Charcot-Marie-Tooth disease(CMT)on endocytosis. In endocytosis the membrane curves in to the cell and is pinched off to form very small (approximately one ten thousandth of a millimetre) ball shaped structures called vesicles, with the cargo inside. The team are currently investigating two types of endocytosis: clathrin-mediated and caveolin-dependent. They are named for the key proteins stabilizing the shape of vesicles and take up different types of cargo. Both require dynamin 2 to function.
Along with important laboratory techniques the PhD student has learnt how to operate state of the art correlative scanning ion conductance microscope and fluorescence confocal microscope (SICM-FCM) setup. The student uses this to look at how single endocytic vesicles form in skin cells from people with centronuclear myopathy and CMT. The researchers found that mutations in different parts of dynamin 2 have different damaging effects on how clathrin-coated vesicles form. Some of the team’s findings are different from results from other researchers. Dr Shevchuk and his student are currently using more techniques to investigate in greater detail how the mutations in dynamin 2 affect endocytosis.
This year the student has also learnt how to follow the formation of individual caveoli vesicles using correlative SICM-FCM. The researchers have also used SICM techniques to measure the stiffness of skin cells because this may be regulated partly by caveolin and clathrin.
Next the team will continue their in-depth analysis of clathrin- dependent endocytosis mechanisms in cells with dynamin 2 mutations. The researchers will look to see if the caveolin-dependent vesicles are affected in skin cells with the dynamin 2 mutations with SICM-FCM and if the stiffness of the cells with the dynamin 2 mutations is different from healthy cells.
If you would like further details about this research project, please email MDUK’s Research team at firstname.lastname@example.org