Periodic Reporting for period 1 - SILIA (Solid Immersion Lens Microscopy to Study Cilia Assembly)
Periodo di rendicontazione: 2019-05-04 al 2021-05-03
The second aim involved studying biological processes in the ciliary tip domain. There are several outstanding questions associated with the ciliary tip, including its role in length regulation of the cilium. It has been known that cilium elongates exclusively at its distal tip. One of proposed hypotheses assumes that the protein complex formed at the tip of mature cilia is involved in its stabilization and prevents an indefinite growth by blocking addition of new tubulin dimers. To explore this hypothesis, an in vitro reconstitution system based on flagellar skeletons was established. Using wild-type and knock-down mutant cell lines it was shown that the tip of mature cilia is indeed incompetent of stimulating microtubule growth. Subsequently, a role of various tip complex constituents in this process was tested using skeletons prepared of depletion mutants.
In living cells, all building blocks are delivered to the tip by intraflagellar transport. However, it remains unclear how are the intraflagellar transport trains turned around at the tip to return back to the cell body. Using the newly developed system, the intraflagellar transport events at the tip were described with unprecedented accuracy. The scientific results have been communicated in a forthcoming methodical research paper and during an invited conference talk.
Besides the research goals, an important part of the project was to broaden researcher’s expertise in various fields of research and technology. Originally trained as a cell biologist, the researcher benefited tremendously from aspects of this project that included optics and engineering (design and construction of the microscope) and software development and machine learning (analysis of the microscopic data). Together with acquired experience in project planning and management, this action provided the researcher with a valuable set of skills needed for his own independent research career.
In order to present this project to broader audience and promote general interest in cell biology and microscopy, the researcher participated in the The European Researchers' Night and gave a talk aimed at high school students interested in microscopy.