Microelectrode Biosensor for Real-Time Measurement of ATP in Biological Tissue

Llaudet, Enrique; Hatz, Sonja; Droniou, Magali; Dale, Nicholas

doi:10.1021/ac048106q

Cited by 254 publications

(238 citation statements)

References 38 publications

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“…It is based on an ATP biosensor built by coating a platinum microelectrode with an ultra thin layer of various enzymes [49,50]. Different enzymes and coating protocols have successfully been used to produce functional, robust and fast-acting ATP electrodes [49,50].…”

Section: The Amperometric Atp Biosensor Microelectrodementioning

confidence: 99%

“…In the presence of glycerol, ATP leads to GK-dependent formation of glycerol-3-phosphate, which together with O 2 is converted to H 2 O 2 and glycerone phosphate by G3P. Subsequently, H 2 O 2 is oxidised at the platinum electrode (+500 mV) which provides a current signal directly proportional to the amount of consumed ATP [50]. ATP-induced current signals are linear over the physiologically relevant ATP concentration range (200 nM to 50 µM) and the microelectrode appears to be very sensitive:~250 mA M −1 cm −2 .…”

Section: The Amperometric Atp Biosensor Microelectrodementioning

confidence: 99%

“…It has been used to detect ATP release during locomotor activity of the spinal cord of Xenopus embryos. It does not detect other nucleotides but may perceive disturbances from other electro-active compounds like 5HT or ascorbate [50]. Using appropriate controls with the uncoated non-sensor electrode or without glycerol, it is possible to verify that the current signals are ATP-specific.…”

Section: The Amperometric Atp Biosensor Microelectrodementioning

confidence: 99%

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ATP release from non-excitable cells

Praetorius

Leipziger

2009

Purinergic Signalling

207

229

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All cells release nucleotides and are in one way or another involved in local autocrine and paracrine regulation of organ function via stimulation of purinergic receptors. Significant technical advances have been made in recent years to quantify more precisely resting and stimulated adenosine triphosphate (ATP) concentrations in close proximity to the plasma membrane. These technical advances are reviewed here. However, the mechanisms by which cells release ATP continue to be enigmatic. The current state of knowledge on different suggested mechanisms is also reviewed. Current evidence suggests that two separate regulated modes of ATP release co-exist in nonexcitable cells: (1) a conductive pore which in several systems has been found to be the channel pannexin 1 and (2) vesicular release. Modes of stimulation of ATP release are reviewed and indicate that both subtle mechanical stimulation and agonist-triggered release play pivotal roles. The mechano-sensor for ATP release is not yet defined.

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Section: The Amperometric Atp Biosensor Microelectrodementioning

confidence: 99%

Section: The Amperometric Atp Biosensor Microelectrodementioning

confidence: 99%

Section: The Amperometric Atp Biosensor Microelectrodementioning

confidence: 99%

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ATP release from non-excitable cells

Praetorius

Leipziger

2009

Purinergic Signalling

207

229

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“…Our particular approach has been to develop microelectrode biosensors for the purines [35,36]. While there are many ways biosensors can operate (indeed luciferase, patch sniffing, and recombinant receptors are all biosensing approaches for the detection of purines), we have developed electrochemical biosensors based around oxidases to provide a real-time signal for both ATP and adenosine and downstream purines.…”

Section: Microelectrode Biosensorsmentioning

confidence: 99%

Measurement of purine release with microelectrode biosensors

Dale

Frenguelli

2011

Purinergic Signalling

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Purinergic signalling departs from traditional paradigms of neurotransmission in the variety of release mechanisms and routes of production of extracellular ATP and adenosine. Direct real-time measurements of these purinergic agents have been of great value in understanding the functional roles of this signalling system in a number of diverse contexts. Here, we review the methods for measuring purine release, introduce the concept of microelectrode biosensors for ATP and adenosine and explain how these have been used to provide new mechanistic insight in respiratory chemoreception, synaptic physiology, eye development and purine salvage. We finish by considering the association of purine release with pathological conditions and examine the possibilities that biosensors for purines may one day be a standard part of the clinical diagnostic tool chest.Keywords Biosensor . Real-time measurement . Epilepsy . Stroke . Chemosensory mechanisms . DevelopmentThe need for real-time purine measurementThe traditional paradigm of synaptic transmission involves exocytotic release of transmitter from a defined presynaptic structure into the narrow gap of the synaptic cleft where it diffuses to activate receptors on the closely apposed postsynaptic membrane. Release is time-locked to the presynaptic action potential and results in the predictable occurrence of postsynaptic events on defined time scales. While capturing the essence of synaptic transmission, this paradigm is an oversimplification. For example, transmitters such as GABA [1] and glutamate [2] can spill out of the synaptic cleft to have more diffuse and widespread actions than this classical model suggests. Under pathological conditions, transmitters and neurotransmitters, such as glutamate [3], can be released by the reversal of Na + -dependent concentrative transporters.The purines, ATP and adenosine, depart from this paradigm even more comprehensively. ATP can indeed be released at synapses [4,5]. However, in the CNS, it seems that it acts mainly as a cotransmitter [6,7], and there are very few synapses where it acts as the principal transmitter. ATP is also released by glial cells [8,9] for the most part via exocytosis [10,11] but see [12]. However, ATP can also be released via a variety of channel-mediated mechanisms (for example, via gap junction hemichannels [13][14][15] and volume-activated channels [16,17]). Receptors for ATP are widespread throughout the CNS; yet, the cellular sources of ATP release to activate these receptors are not immediately obvious. In addition, although ATP can be broken down to ADP and further to adenosine, these breakdown products are themselves active at particular receptor subtypes.The routes for adenosine release and the cellular sources seem to be even more mysterious than those for ATP. For example, there is abundant evidence that adenosine can arise from the breakdown of previously released ATP [18].

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“…Dale's group reported on a microelectrode biosensor for in vivo measurement of ATP in biological tissues. 151 Amperometric biosensors for ATP are of great promise due to the experimentally simplicity. However, most of ATP biosensors with hexokinase and glucose oxidase suffer from the interference of ambient glucose and can only detect ATP in a low and narrow concentration range (200-300 nM) below the physiological concentration in the nervous system and other tissues.…”

Section: Other Enzyme Biosensorsmentioning

confidence: 99%