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Name of scholarship/program

Understanding the cellular and molecular basis for increased fluorescein interactions with epithelial cells after exposure to toxins or irritants (CASE Studentship)

Important description
Principal Supervisor: Curtis Dobson. Co-Supervisors: Carole Maldonado-Codina, Philip Morgan

This BBSRC industrial CASE studentship is a collaboration with Sauflon Pharmaceuticals Limited; it aims to increase our understanding of the cellular and molecular events underlying the interaction of the commonly-used stain fluorescein with cells, particularly after exposure to toxins.

Despite the widespread use of fluorescein in labelling compounds for basic research, its interactions with cells are surprisingly poorly understood. In contact with ocular tissues, fluorescein can produce irregular ‘punctate’ staining that is increased in the presence of toxins like benzalkonium chloride, anaesthetic eye drops, alkalis and contact lens multi-purpose solutions (MPS); the unknown significance of this is of concern to the ophthalmics industry. We have shown an appreciable subset of cells within fluorescein-treated cell cultures shows hyperfluorescence due to internalization of fluorescein, with MPS exposure increasing this. The presence of hyperfluoresent cells is temperature dependent, suggesting an active process. Moreover, hyperfluorescent cells are not necrotic, and release fluorescein more readily at 37˚C, and are capable of cell division. We conclude that fluorescein hyperfluorescence takes place in living cells through active processes, and does not reflect increased staining within dead cells.

The project has two aims. We will (a) gain greater insight into the cell biology underlying hyperfluorescence by visualizing fluorescein entry into diverse cell types, testing whether hyperfluorescent cells are undergoing apoptosis, investigating which organelles concentrate fluorescein and using FACS to separate and compare the biochemistry and cell biology of hyperfluorescent and non-hyperfluorescent cells; and (b) we will dissect which MPS components cause hyperfluorescence, and test how this is mediated by lens materials and solution components.

The successful candidate will join a multi-award winning research group with access to state of the art microscopy and cell biology techniques. The project tackles a fundamental biological question of significance to a multi-billion dollar industry with the direct involvement of a major company operating in this sector.

Eligibility and other criteria
This research project has funding attached. Funding for this project is available to citizens of a number of European countries (including the UK). In most cases this will include all EU nationals. However full funding may not be available to all applicants and you should read the full department and project details for further information.

Application deadline
*Applications accepted all year round

Additional information, and important URL
For full information on how to apply for this project, please visit www.manchester.ac.uk/phdprogrammes

Applications are invited from UK/EU students only. Applicants must have obtained, or be about to obtain, at least an upper second class honours degree in a relevant subject.

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