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Building and breaking epithelial organs: an optogenetic approach

Supervisor: Clare Buckley

Co-supervisor: Magdalena Zernicka-Goetz

My lab uses cutting edge optogenetic and live confocal imaging approaches within the whole zebrafish neural tube to manipulate the polarity of single cells using light. In combination with CRISPR-mediated functional knock down experiments, this allows us to ask how cell polarity is involved in building organs during development and breaking them during diseases such as carcinoma.

Up until now it has been challenging to design experiments to test the sufficiency of polarity proteins to drive tissue organisation. To solve this problem, we recently published an in vivo optogenetic technique that allows the use of light to reversibly localise proteins and signalling to specific subcellular positions with unprecedented levels of spatiotemporal control. Critically, this approach allows us to test the importance of protein LOCATION, rather than just the necessity of protein function.

There are several projects available within the lab. For example:

1. How are cell polarity and division linked during development?

During epithelial establishment, cell division and polarity are known to be strongly linked but our recent work discovered a novel mechanism of division-independent de novo cell polarisation. We will further explore the interrelationship between cell polarity and cell division in the context of a whole organ by using optogenetics to move key polarity and division regulators and characterising the downstream cellular consequences via live imaging.

2. What is the role of polarity dysregulation in tissue disruption?

There is emerging evidence that polarity defects might be upstream of tissue disorganisation at the onset of diseases such as cancer. However, it is still unclear whether polarity defects are a cause or consequence of tissue disorganisation. We will test the role of polarity dysregulation in tissue disruption by optogenetically manipulating polarity-linked signalling pathways such as the cancer-linked PI3K pathway in individual cells within the whole neural tube.

 

References

Buckley et al. 2016. Dev Cell. Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo. https://www.ncbi.nlm.nih.gov/pubmed/26766447

Buckley et al. 2014. Semin Cell Dev Biol. Establishing the plane of symmetry for lumen formation and bilateral brain formation in the zebrafish neural rod. https://www.ncbi.nlm.nih.gov/pubmed/24721474

Buckley et al. 2013. EMBO J. Mirror-symmetric microtubule assembly and cell interactions drive lumen formation in the zebrafish neural rod. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3545300/.