Dr E Rosato, Prof C P Kyriacou
Applications accepted all year round
Self-Funded PhD Students Only
About the Project
The circadian clock controls every facet of behaviour and physiology, and is therefore a very important factor in human and animal health and well-being. In fruit flies and mammals, the same genes appear to play similar roles in determining how the 24 h clock works [reviewed in 1]. However, the ease of genetic analysis and the molecular tools available in the fruit fly Drosophila melanogaster, means that progress is particularly rapid with this organism. We are interested in a blue-light sensitive protein called CRYPTOCHROME (CRY), which employs light energy for signalling. The main function of CRY in the fly brain, is to bring light information directly into the clock mechanism, resetting it. However, our work and that of others point to additional roles for CRY [2]. One of these results in a light-dependent increase in neuronal firing due to an indirect interaction with Shaker, a potassium channel [3]. Additionally, we have unpublished evidence that CRY is important during development to set the “connectivity” among several clock neurons, namely to determine their level of interaction. This role is light-independent and we currently do not understand how it is achieved. We have generated transgenic fly lines carrying mutations in the C-terminus of CRY, which is important for signalling. We have measured behavioural differences among those lines and we want to identify the molecular and cellular mechanisms responsible. We will use a combination of approaches including genetics (crossing of mutants and transgenic lines), molecular biology (generation of specific mutants using gene editing), biochemistry (immunoprecipitation and mass spectrometry), behaviour (locomotor activity rhythms) and imaging (measurements of cell activation and connectivity) to investigate how CRY modulates the interaction among neurons and which additional molecules take part in the process. This project does not require previous experience with Drosophila but strong commitment and confidence in using different techniques (training provided) are essential.
Funding Notes
This is a self-funded project.
References
. Ozkaya & Rosato (2012) The circadian clock of the fly: a neurogenetics journey through time. Advances in Genetics, 77: 79-123.
2. Dissel S, Hansen CN, Ozkaya O, Hemsley M, Kyriacou CP, Rosato E. (2014) The logic of circadian organization in Drosophila, Curr Biol, 24: 2257-2266
3. Fogle KJ, Baik LS, Houl JH, Tran TT, Roberts L, Dahm NA, Cao Y, Zhou M, Holmes TC (2015). CRYPTOCHROME-mediated phototransduction by modulation of the potassium ion channel β-subunit redox sensor. PNAS,112: 2245-50.