Role of vesicular nucleotide transporter VNUT (SLC17A9) in release of ATP from AR42J cells and mouse pancreatic acinar cells
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ATP is released from cells in response to various stimuli. Our previous studies on pancreas indicated that pancreatic acini could be major stores of secreted ATP. In the present study, our aim was to establish the role of the vesicular nucleotide transporter (VNUT), SLC17A9, in storage and release of ATP. Freshly prepared acini from mice and AR42J rat acinar cells were used in this study. We illustrate that in AR42J cells, quinacrine (an ATP store marker) and Bodipy ATP (a fluorescent ATP analog) co-localized with VNUT-mCherry to vesicles/granules. Furthermore, in acini and AR42J cells, a marker of the zymogen granule membranes, Rab3D, and VNUT co-localized. Dexamethasone treatment of AR42J cells promoted formation of acinar structures, paralleled by increased amylase and VNUT expression, and increased ATP release in response to cholinergic stimulation. Mechanical stimulus (pressure) and cell swelling also induced ATP release, but this was not influenced by dexamethasone, most likely indicating different non-zymogen-related release mechanism. In conclusion, we propose that VNUT-dependent ATP release pathway is associated with agonist-induced secretion process and downstream purinergic signalling in pancreatic ducts.
KeywordsPancreas ATP release VNUT SLC17A9 AR42J Pancreatitis Mechanical stress
vesicular nucleotide transporter
Imaging was performed at the Center for Advanced Bioimaging (CAB) Denmark, University of Copenhagen, and support from N.M. Christensen is greatly appreciated. The technical assistance of Pernille Roshof and David Sørensen is greatly acknowledged.
This project was supported by The Danish Council for Independent Research | Natural Sciences. Part of instrumentation was supported by The Lundbeck Foundation, The Novo Nordisk Foundation, and The Carlsberg Foundation.
Conflict of interest
There are no conflicts of interest, financial or otherwise.
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