Isolation and molecular characterisation of primary cilia from neural stem cells

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The ability to exit the cell cycle and undergo quiescence is key to protect the genetic information and maintain the function of adult stem cells. Therefore, the understanding of molecular and cel-lular mechanisms underlying the regulation of stem cell quiescence is a fundamental issue in the field. In this context, primary cilia, organelles on the surface of stem cells, are of special interest due to their essential role in signal transduction and in the regulation of cell cycle progression. The main aim of the current research project is to purify and fully characterize signalling components in primary cilia from neural stem cells (NSCs). To this end we will take advantage of mechanical, chemical and genetic approaches to detach and label primary cilia from neural precursors and purify primary cilia from the cell body of NSCs using flow cytometry. Defining the molecular composition of primary cilia in NSCs will provide new tools for the analysis of cilia function and diversity. It may also have clinical relevance, as it will provide new insights into the molecular mechanisms underlying the regulation of NSC quiescence essential to release their regenerative potential.