How to build and maintain a 3-dimensional polarised epithelial sheet

We are interested in how cells maintain normal cell shape within an epithelium. Correct epithelial cell shape is essential to maintain tissue integrity and when deregulated can cause diseases such as malignant cancers, diabetes, inflammation, and aging. This project offers a PhD student the exciting opportunity to work with three collaborating research groups, using state-of-the-art techniques, and using in vivo and in vitro systems, to better understand the molecular mechanisms underlying cell and tissue morphology.

Our recent work, using the living fruit fly as a model system, has begun to unravel the complex intracellular molecular machinery required for cells to take their characteristic shape and to correctly form and position sub-cellular structures, such as cell-cell junctions and dynamic protrusions. The PhD student will take this research further by working with three research groups in three Schools within the University of Nottingham: Dr Georgiou (Life Sciences) has developed tools that enable the labelling and manipulation of individual epithelial cells within the living fly. Dr Ashworth (Veterinary Medicine) has developed an advanced 3D in vitro culture system, allowing independent control over extra-cellular matrix (ECM) stiffness and composition. Dr Mendonca (Engineering) has developed ‘OptoRheo’, an optical instrument that enables non-destructive measurement of ECM stiffness at the microscale while also capturing corresponding live cellular dynamics using 3D light-sheet fluorescence microscopy. By combining the expertise of these groups, we aim to: (i) apply advanced imaging to our in vivo model using a customised light-sheet fluorescence microscope, developed by Dr Mendonca, to image cell morphology and dynamic protrusions in high temporal and spatial resolution in the living animal; (ii) transfer technology to in vitro models, using advanced biomaterials, to understand how properties of human stroma affect epithelial architecture and behaviour. We plan to use these complementary technologies to investigate the roles of ECM composition and stiffness in determining epithelial cell shape and tissue architecture. This will allow the link between epithelial sheet development and intracellular signalling to be explored in a system allowing precise tuning of ECM properties.

This project offers the student a fantastic opportunity to learn state-of-the-art techniques, including:

·      Sophisticated fly genetics

·      Live high-resolution imaging using confocal and light-sheet microscopy

·      Fly pupal mounting and dissections

·      Immunohistochemistry

·      Advanced 3D cell culture

·      Use of specialised imaging software

·      A wide variety of molecular biology techniques

·      Data analysis

·      Statistical analysis and numeracy skills