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Monitoring of ATP release from individual cells using a biosensor approach

, : Monitoring of ATP release from individual cells using a biosensor approach. Biologicheskie Membrany (Moscow) 24(4): 309-315

Adenosine triphosphate (ATP) is a neurotransmitter/neuromodulator in both central and peripheral nervous systems. Particularly in the taste bud, a peripheral taste organ, ATP serves as an afferent neurotransmitter. To examine the mechanism mediating ATP secretion in taste cells, here we elaborated an approach for assaying ATP in extracellular medium by employing a biosensor, that is, cells responsive to ATP. Two lines of ATP-sensitive cells, HEK-293 and COS-1, which both endogenously express P2Y receptors, were employed. In addition, HEK-293 cells transfected with P2X(3) receptors were also used. By most of relevant parameters (threshold response, inactivation kinetics of ATP responses, and refractory period) COS-1 cells were more suitable as an ATP sensor than HEK-293 cells, both original and transfected. For HEK-293 cells, one of pitfalls was that they were highly responsive to mechanical disturbances, e.g. solution flux elicited by application of a chemical stimulus, Owing to the expression of mechanosensitive Ca2+-permeable cation channels. In COS-1 cells, ATP-dependent Ca2+ transients were generated mostly due to Ca2+ release, the feature allowing one to control the activity of ATP-releasing cells electrophysiologically and to monitor their ATP secretion by Ca2+ responses of the ATP-biosensor. By using this technique, it was demonstrated that individual taste cells of the mouse released ATP in response to their depolarization.


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