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Using Amperometric, Enzyme-Based Biosensors for Performing Longitudinal Measurements of Extracellular Adenosine 5-Triphosphate in the Mouse

  • Edward BeamerEmail author
  • Tobias Engel
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2041)

Abstract

Adenosine 5-triphosphate (ATP) functions in the central nervous system as an extracellular signaling molecule. While much progress has been made in understanding the circumstances under which it is released, from in vitro preparations, in vivo has proven more challenging. Microdialysis followed by high-performance liquid chromatography has been employed to demonstrate a spike in extracellular concentrations under some pathological conditions including seizures, but this method lacks the sensitivity to detect extracellular ATP at concentrations present under normal physiological conditions. An alternative approach, the use of amperometric, enzyme-based microelectrode biosensors for measuring extracellular ATP in vivo have been employed in the rabbit. Here, we describe a protocol for measuring ATP concentrations using these biosensors in the mouse, simultaneously with electroencephalogram recordings. This approach is ideal for investigating the relationship between ATP release and seizures.

Key words

Microelectrode biosensors Amperometric detection Enzyme-based Adenosine-5-triphosphate In vivo electrochemistry 

Notes

Acknowledgments

This work was supported by funding from the Health Research Board (HRA-POR-2015-1243 to T.E.); Science Foundation Ireland (13/SIRG/2098 and 17/CDA/4708 to T.E.), from the H2020 Marie Skłowdowksa-Curie Actions Individual Fellowship (753527 to E.B.) and from the European Union’s Horizon 2020 research and innovation program under the Marie Sklowdowska-Curie grant agreement (No. 766124 to T.E.).

References

  1. 1.
    Langen P, Hucho F (2008) Karl Lohmann and the discovery of ATP. Angew Chem Int Ed Engl 47:1824–1827CrossRefGoogle Scholar
  2. 2.
    Holton FA, Holton P (1954) The capillary dilator substances in dry powders of spinal roots; a possible role of adenosine triphosphate in chemical transmission from nerve endings. J Physiol 126:124–140CrossRefGoogle Scholar
  3. 3.
    Burnstock G (1972) Purinergic nerves. Pharmacol Rev 24:509–581PubMedGoogle Scholar
  4. 4.
    Burnstock G, Krugel U, Abbracchio MP, Illes P (2011) Purinergic signalling: from normal behaviour to pathological brain function. Prog Neurobiol 95:229–274CrossRefGoogle Scholar
  5. 5.
    Dombrowski KE, Ke Y, Brewer KA, Kapp JA (1998) Ecto-ATPase: an activation marker necessary for effector cell function. Immunol Rev 161:111–118CrossRefGoogle Scholar
  6. 6.
    Franke H, Grummich B, Hartig W, Grosche J, Regenthal R, Edwards RH, Illes P, Krugel U (2006) Changes in purinergic signaling after cerebral injury — involvement of glutamatergic mechanisms? Int J Dev Neurosci 24:123–132CrossRefGoogle Scholar
  7. 7.
    Melani A, Turchi D, Vannucchi MG, Cipriani S, Gianfriddo M, Pedata F (2005) ATP extracellular concentrations are increased in the rat striatum during in vivo ischemia. Neurochem Int 47:442–448CrossRefGoogle Scholar
  8. 8.
    Frenguelli BG, Wigmore G, Llaudet E, Dale N (2007) Temporal and mechanistic dissociation of ATP and adenosine release during ischaemia in the mammalian hippocampus. J Neurochem 101:1400–1413CrossRefGoogle Scholar
  9. 9.
    Wu PH, Phillis JW (1978) Distribution and release of adenosine triphosphate in rat brain. Neurochem Res 3:563–571CrossRefGoogle Scholar
  10. 10.
    Frenguelli BG, Wall MJ (2016) Combined electrophysiological and biosensor approaches to study purinergic regulation of epileptiform activity in cortical tissue. J Neurosci Methods 260:202–214CrossRefGoogle Scholar
  11. 11.
    Lietsche J, Imran I, Klein J (2016) Extracellular levels of ATP and acetylcholine during lithium-pilocarpine induced status epilepticus in rats. Neurosci Lett 611:69–73CrossRefGoogle Scholar
  12. 12.
    Dona F, Conceicao IM, Ulrich H, Ribeiro EB, Freitas TA, Nencioni AL, Da Silva Fernandes MJ (2016) Variations of ATP and its metabolites in the hippocampus of rats subjected to pilocarpine-induced temporal lobe epilepsy. Purinergic Signal 12:295–302CrossRefGoogle Scholar
  13. 13.
    Picher M, Burch LH, Boucher RC (2004) Metabolism of P2 receptor agonists in human airways: implications for mucociliary clearance and cystic fibrosis. J Biol Chem 279:20234–20241CrossRefGoogle Scholar
  14. 14.
    Beamer E, Goloncser F, Horvath G, Beko K, Otrokocsi L, Kovanyi B, Sperlagh B (2016) Purinergic mechanisms in neuroinflammation: an update from molecules to behavior. Neuropharmacology 104:94–104CrossRefGoogle Scholar
  15. 15.
    Masino SA, Kawamura M Jr, Ruskin DN (2014) Adenosine receptors and epilepsy: current evidence and future potential. Int Rev Neurobiol 119:233–255CrossRefGoogle Scholar
  16. 16.
    Pangrsic T, Potokar M, Stenovec M, Kreft M, Fabbretti E, Nistri A, Pryazhnikov E, Khiroug L, Giniatullin R, Zorec R (2007) Exocytotic release of ATP from cultured astrocytes. J Biol Chem 282:28749–28758CrossRefGoogle Scholar
  17. 17.
    Brown P, Dale N (2002) Spike-independent release of ATP from Xenopus spinal neurons evoked by activation of glutamate receptors. J Physiol 540:851–860CrossRefGoogle Scholar
  18. 18.
    Gourine AV, Dale N, Llaudet E, Poputnikov DM, Spyer KM, Gourine VN (2007) Release of ATP in the central nervous system during systemic inflammation: real-time measurement in the hypothalamus of conscious rabbits. J Physiol 585:305–316CrossRefGoogle Scholar
  19. 19.
    Dale N, Frenguelli BG (2009) Release of adenosine and ATP during ischemia and epilepsy. Curr Neuropharmacol 7:160–179CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Physiology and Medical PhysicsRoyal College of Surgeons in IrelandDublin 2Ireland

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