Controlled release of ATP can be used for understanding extracellular purinergic signaling. While coarse mechanical forces and hypotonic stimulation have been utilized in the past to initiate ATP release from cells, these methods are neither spatially accurate nor temporally precise. Further, these methods cannot be utilized in a highly effective cell-specific manner. To mitigate the uncertainties regarding cellular-specificity and spatio-temporal release of ATP, we herein demonstrate use of optogenetics for ATP release. ATP release in response to optogenetic stimulation was monitored by Luciferin-Luciferase assay (North American firefly, photinus pyralis) using luminometer as well as mesoscopic bioluminescence imaging. Our result demonstrates repetitive release of ATP subsequent to optogenetic stimulation. It is thus feasible that purinergic signaling can be directly detected via imaging if the stimulus can be confined to single cell or in a spatially-defined group of cells. This study opens up new avenue to interrogate the mechanisms of purinergic signaling.

Date Published: 8 March 2013
PDF: 7 pages
Proc. SPIE 8586, Optogenetics: Optical Methods for Cellular Control, 85860L (8 March 2013); doi: 10.1117/12.2008234

Published in SPIE Proceedings Vol. 8586:
Optogenetics: Optical Methods for Cellular Control
Samarendra K. Mohanty; Nitish V. Thakor, Editor(s)