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Advanced mass spectrometry analysis of metabolite flux through the purine salvage pathway in brain

Principal Supervisor: Professor Bruno Frenguelli, School of Life Sciences

Co-supervisor: Dr Alex Jones (SLS) & Dr Andrew Bottrill (Proteomics RTP)

PhD project title: Advanced mass spectrometry analysis of metabolite flux through the purine salvage pathway in brain

University of Registration: University of Warwick

Project outline:

All cells require ATP as an energy source to survive. This is especially true for brain cells where there is little by way of energy stores such as glycogen. ATP is primarily generated from ADP from mitochondria, but in brain cells, the primary source of the ADP arises from the purine salvage pathway, a cytosolic biochemical pathway that converts ATP metabolites initially back into AMP from which ADP is synthesised. This is a problem for the brain when it is injured as these metabolites are lost from the brain and thus unavailable for salvage.

We have shown that providing the salvage pathway with exogenous substrates (ribose and adenine; “RibAde”) allows injured brain tissue to restore ATP levels to normal levels. We have also shown that this increased reservoir of ATP translates to great release of adenosine during physiological and pathological conditions where it exerts powerful and important protective actions on brain electrical activity. Indeed, RibAde protected greater brain tissue and accelerated recovery in an animal model of stroke.

In this project we will use advanced mass spectrometry techniques to identify the fate of ribose and adenine as they cycle through the purine salvage pathway. By using stable heavy isotopes of carbon (for ribose) and nitrogen (for adenine) we will be able to conclusively identify the intracellular molecules into which they are incorporated. In addition, and in collaboration with Dr Martin Lochner (University of Bern), we will utilise novel chemotype approaches to the elevation of cellular ATP, which in the long-run may be beneficial in the treatment of acute brain injuries.

References:

  1. Frenguelli BG. (2017) The Purine Salvage Pathway and the Restoration of Cerebral ATP: Implications for Brain Slice Physiology and Brain Injury. Neurochem Res. doi: 10.1007/s11064-017-2386-6.
  2. Faller et al., (2017) Proof of concept and feasibility studies examining the influence of combination ribose, adenine and allopurinol treatment on stroke outcome in the rat Brain & Neurosci Adv doi.org/10.1177/2398212817717112
  3. zur Nedden S, et al., (2014) Modulation of intracellular ATP determines adenosine release and functional outcome in response to metabolic stress in rat hippocampal slices and cerebellar granule cells. J Neurochem. 128: 111-24 doi:10.1111/jnc.12397
  4. zur Nedden S et al., (2011). Intracellular ATP influences synaptic plasticity in area CA1 of rat hippocampus via metabolism to adenosine and activity-dependent activation of adenosine A1 receptors. J Neurosci. 31: 6221-34 doi:10.1523/JNEUROSCI.4039-10

BBSRC Strategic Research Priority: Integrated Understanding of Health: Ageing & Understanding the Rules of Life:Neuroscience and behaviour & Pharmaceuticals

Techniques that will be undertaken during the project:

  • Primary neuronal culture and hippocampal slice culture
  • Brain slice preparation
  • Electrophysiology
  • Microelectrode biosensors
  • Mass spectrometry
  • Quantitative biology
  • Organic chemistry and the synthesis of novel compounds

Contact: Professor Bruno Frenguelli, University of Warwick