Imaging the dynamics of centriole biogenesis beyond the diffraction limit of light.

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Poster Session Two
Dr Alan Wainman (1), Professor Jordan Raff (1)
1. University of Oxford

live imaging, dynamics.

Abstract text

Centrosomes are cellular organelles that constitute the major microtubule organising centre in animal cells and play important roles in the regulation of the cell cycle. Each centrosome is formed of two centrioles surrounded by the pericentriolar material, from which microtubules are nucleated. To ensure that, upon cell division, each daughter cell inherits one centrosome with two centrioles, during S phase the two centrioles separate (disengagement) and, each centriole becomes a ‘mother’ centriole that templates the formation of a ‘daughter’ centriole (duplication). Daughter centrioles then grow, perpendicular to the mother, during interphase. This pair of centrioles in a centrosome, moves to opposite sides of the cell during mitosis in order to form the bipolar mitotic spindle. After cell division, each resulting cell inherits one centrosome containing two centrioles.


Although centrioles have been studied for many years, two important questions remain unanswered a) what are the dynamics of the centriole disengagement process and b) when and exactly where are the various molecular components that form the centrioles dynamically incorporated as centrioles duplicate. As individual centrioles are just below the diffraction limit, traditional imaging techniques had, until now, been insufficient to tackle these questions. 


Here, we describe a series of imaging studies investigating these two questions. We use the Drosophila embryo, as this system offers a series of advantages for quantitative dynamic imaging experiments, including short cell cycles, synchronously dividing centrosomes, and their cortical localisation close to the coverslip. We combine the fruit fly system with quantitative and super resolution techniques to provide new insights into the nature of centriole growth.