Abstract
The function of alpha-cells in patients with type 2 diabetes is often disturbed; glucagon secretion is increased at hyperglycaemia, yet fails to respond to hypoglycaemia. A crucial mechanism behind the fine-tuned release of glucagon relies in the exocytotic machinery including SNARE proteins. Here, we aimed to investigate the temporal role of syntaxin 1A and SNAP-25 in mouse alpha-cell exocytosis. First, we used confocal imaging to investigate glucose dependency in the localisation of SNAP-25 and syntaxin 1A. SNAP-25 was mainly distributed in the plasma membrane at 2.8 mM glucose, whereas the syntaxin 1A distribution in the plasma membrane, as compared to the cytosolic fraction, was highest at 8.3 mM glucose. Furthermore, following inclusion of an antibody against SNAP-25 or syntaxin 1A, exocytosis evoked by a train of ten depolarisations and measured as an increase in membrane capacitance was reduced by ~50%. Closer inspection revealed a reduction in the refilling of granules from the reserve pool (RP), but also showed a decreased size of the readily releasable pool (RRP) by ~45%. Disparate from the situation in pancreatic beta-cells, the voltage-dependent Ca2+ current was not reduced, but the Ca2+ sensitivity of exocytosis decreased by the antibody against syntaxin 1A. Finally, ultrastructural analysis revealed that the number of docked granules was >2-fold higher at 16.7 mM than at 1 mM glucose. We conclude that syntaxin 1A and SNAP-25 are necessary for alpha-cell exocytosis and regulate fusion of granules belonging to both the RRP and RP without affecting the Ca2+ current.
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Acknowledgements
We thank Britt-Marie Nilsson, Anna-Maria V Ramsay and Kristina Borglid for technical assistance. This work was supported by the Swedish Research Council, The Novo Nordisk Foundation, The Swedish Diabetes Association, Knut and Alice Wallenberg Foundation, The Albert Påhlsson Foundation, UMAS Foundation and the EU through a Marie Curie Intra-European Fellowship (MGP). LE is a senior researcher at the Swedish Research Council.
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Andersson, S.A., Pedersen, M.G., Vikman, J. et al. Glucose-dependent docking and SNARE protein-mediated exocytosis in mouse pancreatic alpha-cell. Pflugers Arch - Eur J Physiol 462, 443–454 (2011). https://doi.org/10.1007/s00424-011-0979-5
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DOI: https://doi.org/10.1007/s00424-011-0979-5