Using Amperometric, Enzyme-Based Biosensors for Performing Longitudinal Measurements of Extracellular Adenosine 5-Triphosphate in the Mouse
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 wordsMicroelectrode biosensors Amperometric detection Enzyme-based Adenosine-5-triphosphate In vivo electrochemistry
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.).