Voltage-gated ion channels are highly conserved from worms to man, yet the detail of the neurophysiological functions of many of these channels remains mysterious. Various members of this broad family have been linked to oxygen sensing, but their roles and mechanism of action are not well understood.
The aim of this project is to achieve a mechanistic understanding of the channels’ roles in regulating cellular oxygen homeostasis. For this we can take advantage of several novel and distinct phenotypes of conserved ion channels in C. elegans that we discovered, which alter the function of the O2-sensing neural circuit and behavioural responses. We have also identified novel genetic interactions of the channels with signalling factors that tune sensory responses. We aim to approach the question on several fronts, investigating the biophysical properties of target channels, their regulation and molecular interactions, and how these properties enable their role in regulating oxygen responses.
We will shed light on the mechanisms of channel activity by:
1. performing a structure-function analysis of how these channels and their domains sculpt oxygen responses
2. investigating their functional interaction with other signalling proteins in tuning and sustaining O2 responses
3. identifying and dissecting new protein interactors and the pharmacology of the channels in a medium-throughput assay.
We will express full-length and truncated channels in human cultured cell lines and carry out a biophysical characterisation of O2-modulated currents with electrophysiology. We will also co-express interacting proteins and conduct a biophysical investigation of the interaction, determine interaction motifs and test co-localisation and direct binding of the proteins by co-immunoprecipitation. We will assess novel interactors/regulators and their mechanism of interaction with the channels, based on the outcome of an interactor screen in C. elegans conducted by other lab members. We will also explore channel pharmacology in microplate reader assays. We will complement these studies with interrogation of the role of the proteins in C. elegans neural circuits in vivo, using microfluidics chambers to assay O2-evoked behaviour, functional neuronal imaging and confocal microscopy to assess channel function and localisation.
The project gives the opportunity to advance a mechanistic understanding of the physiology and regulation of these medically relevant channels and shed light on O2-sensing mechanisms. There is an unmet need to provide better treatment for diseases and disorders linked to these channels, such as epilepsy, schizophrenia or cancer. Our fundamental research will inform therapeutic approaches dealing with these channels in brain disorders in the longer term. You will benefit from training in widely applicable state-of-the-art techniques in neuroscience, such as electrophysiological recordings and functional interrogation of C. elegans neurons, and the exposure to the cutting-edge research of other Edinburgh neuroscientists.
Prof Mike Shipston
Dr Iain Rowe (Robert Gordon University, Aberdeen)