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Purinergic Signalling

, Volume 10, Issue 3, pp 431–440 | Cite as

Role of vesicular nucleotide transporter VNUT (SLC17A9) in release of ATP from AR42J cells and mouse pancreatic acinar cells

  • K. A. Haanes
  • J. M. Kowal
  • G. Arpino
  • S. C. Lange
  • Y. Moriyama
  • P. A. Pedersen
  • I. NovakEmail author
Original Article

Abstract

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.

Keywords

Pancreas ATP release VNUT SLC17A9 AR42J Pancreatitis Mechanical stress 

Abbreviations

ZG

zymogen granule

VNUT

vesicular nucleotide transporter

Notes

Acknowledgments

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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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • K. A. Haanes
    • 1
  • J. M. Kowal
    • 1
  • G. Arpino
    • 1
  • S. C. Lange
    • 1
  • Y. Moriyama
    • 2
  • P. A. Pedersen
    • 1
  • I. Novak
    • 1
    Email author
  1. 1.Department of Biology, Section Molecular Integrative PhysiologyUniversity of CopenhagenCopenhagenDenmark
  2. 2.Department of Membrane Biochemistry, Graduate School for Medicine, Dentistry and Pharmaceutical SciencesOkayama UniversityOkayamaJapan

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