BIOMAT Database Logo BIOMAT Database Logo

Detecting ATP release by a biosensor method. 2004

Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Department of Cell Physiology, National Institute for Physiological Sciences, Okazaki 444-8585, Japan.

Cells release adenosine 5'-triphosphate (ATP) into the extracellular space in response to various stimuli. This released ATP plays an important physiological role in cell-to-cell signal transduction. The bulk ATP concentration can be detected using a conventional luciferin-luciferase assay. However, the ATP concentration in the vicinity of the cell surface is often different from the bulk concentration because of its rapid degradation by ecto-ATPases and because of delayed diffusion due to unstirred layer effects. Here, we describe a simple biosensor method to measure the local ATP concentration on the cell surface in real time. The method is based on the ATP-dependent opening of ligand-gated cation channels of purinergic P2X receptors expressed in undifferentiated pheochromocytoma (PC12) cells or in human embryonic kidney 293 (HEK293) cells stably transfected with recombinant P2X2 purinergic receptors. Under the whole-cell configuration of patch-clamp, a sensor PC12 cell or HEK293 is positioned within the proximity of a target cell, and the P2X-mediated currents induced by ATP released from a given site on the target cell surface is measured. The ATP release is quantified by a calibration procedure utilizing local puff applications of ATP at preset concentrations.

UI MeSH Term Description Entries
D007668 Kidney Body organ that filters blood for the secretion of URINE and that regulates ion concentrations. Kidneys
D010673 Pheochromocytoma A usually benign, well-encapsulated, lobular, vascular tumor of chromaffin tissue of the ADRENAL MEDULLA or sympathetic paraganglia. The cardinal symptom, reflecting the increased secretion of EPINEPHRINE and NOREPINEPHRINE, is HYPERTENSION, which may be persistent or intermittent. During severe attacks, there may be HEADACHE; SWEATING, palpitation, apprehension, TREMOR; PALLOR or FLUSHING of the face, NAUSEA and VOMITING, pain in the CHEST and ABDOMEN, and paresthesias of the extremities. The incidence of malignancy is as low as 5% but the pathologic distinction between benign and malignant pheochromocytomas is not clear. (Dorland, 27th ed; DeVita Jr et al., Cancer: Principles & Practice of Oncology, 3d ed, p1298) Pheochromocytoma, Extra-Adrenal,Extra-Adrenal Pheochromocytoma,Extra-Adrenal Pheochromocytomas,Pheochromocytoma, Extra Adrenal,Pheochromocytomas,Pheochromocytomas, Extra-Adrenal
D002138 Calibration Determination, by measurement or comparison with a standard, of the correct value of each scale reading on a meter or other measuring instrument; or determination of the settings of a control device that correspond to particular values of voltage, current, frequency or other output. Calibrations
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000255 Adenosine Triphosphate An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter. ATP,Adenosine Triphosphate, Calcium Salt,Adenosine Triphosphate, Chromium Salt,Adenosine Triphosphate, Magnesium Salt,Adenosine Triphosphate, Manganese Salt,Adenylpyrophosphate,CaATP,CrATP,Manganese Adenosine Triphosphate,MgATP,MnATP,ATP-MgCl2,Adenosine Triphosphate, Chromium Ammonium Salt,Adenosine Triphosphate, Magnesium Chloride,Atriphos,Chromium Adenosine Triphosphate,Cr(H2O)4 ATP,Magnesium Adenosine Triphosphate,Striadyne,ATP MgCl2
D000310 Adrenal Gland Neoplasms Tumors or cancer of the ADRENAL GLANDS. Adrenal Cancer,Adrenal Gland Cancer,Adrenal Neoplasm,Cancer of the Adrenal Gland,Neoplasms, Adrenal Gland,Adrenal Cancers,Adrenal Gland Cancers,Adrenal Gland Neoplasm,Adrenal Neoplasms,Cancer, Adrenal,Cancer, Adrenal Gland,Cancers, Adrenal,Cancers, Adrenal Gland,Neoplasm, Adrenal,Neoplasm, Adrenal Gland,Neoplasms, Adrenal
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D014162 Transfection The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES. Transfections
D015374 Biosensing Techniques Any of a variety of procedures which use biomolecular probes to measure the presence or concentration of biological molecules, biological structures, microorganisms, etc., by translating a biochemical interaction at the probe surface into a quantifiable physical signal. Bioprobes,Biosensors,Electrodes, Enzyme,Biosensing Technics,Bioprobe,Biosensing Technic,Biosensing Technique,Biosensor,Electrode, Enzyme,Enzyme Electrode,Enzyme Electrodes,Technic, Biosensing,Technics, Biosensing,Technique, Biosensing,Techniques, Biosensing

Related Publications

Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
A new method for detecting histamine release.
April 1984, Agents and actions,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Detecting Anastasis In Vivo by CaspaseTracker Biosensor.
February 2018, Journal of visualized experiments : JoVE,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Optical ATP biosensor for extracellular ATP measurement.
May 2013, Biosensors & bioelectronics,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Evaluation of a method for detecting metal release from gold; cysteine enhances release.
September 2013, Acta dermato-venereologica,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Microglia release ATP by exocytosis.
August 2013, Glia,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
A Protein-Based Biosensor for Detecting Calcium by Magnetic Resonance Imaging.
September 2021, ACS sensors,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
ATP-stimulated release of ATP by human endothelial cells.
June 1996, Journal of cardiovascular pharmacology,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
K+ depolarization evokes ATP, adenosine and glutamate release from glia in rat hippocampus: a microelectrode biosensor study.
November 2012, British journal of pharmacology,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Detecting Redox Potentials Using Porous Boron Nitride/ATP-DNA Aptamer/Methylene Blue Biosensor to Monitor Microbial Activities.
January 2022, Micromachines,
Seiji Hayashi, and Akihiro Hazama, and Amal K Dutta, and Ravshan Z Sabirov, and Yasunobu Okada
Microglial migration mediated by ATP-induced ATP release from lysosomes.
June 2012, Cell research,
Copied contents to your clipboard!