During development, the genesis of daughter cells with different fates is established by asymmetric cell division, a highly conserved mechanism for the segregation of specific cell-fate determinants.
A central issue in the field of cell biology is to identify specifically the mechanisms that govern cell diversity through asymmetrical partitioning of these determinants. In this project we propose to follow in real time the path, movements and interaction of two key proteins involved in the asymmetric division of Drosophila neuroblasts.
Using single molecule fluorescence microscopy, targeted semiconductor nanocrystals (quantum dots) and various genetic neuroblast mutants, we propose to follow the intracellular routes and determine the spatial and temporal requirements of the cargo protein Miranda and its associated molecular motor Myosin VI.
Through a multidisciplinary approach, we will stoichiometrically label Miranda to quantum dots and develop a GFP-based high affinity molecular marker for the targeting of t he nanoprobes to Myosin VI in the cytoplasm of neuroblasts. Following cell injection, we will perform high speed and multicolour imaging to track the diffusing single proteins in three dimensions.
The developments proposed in this project will not only provide new information on the mechanisms of asymmetric division, but will also offer new approaches and new tools to the fields of optical microscopy, cell biology, and nanomaterials.
Fields of science
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