CFTR and Anoctamin 1 (ANO1) contribute to cAMP amplified exocytosis and insulin secretion in human and murine pancreatic beta-cells

BMC Medicine

Lena Eliasson²  Malin Flodström-Tullberg¹  Anna Wendt²  Jonathan LS Esguerra²  Anna Edlund² 

[1] Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden;Unit of Islet Cell Exocytosis, Lund University Diabetes Centre, Department Clinical Sciences in Malmö, Lund University, Clinical Research Centre, SUS Malmö, Jan Waldenströms gata 35, SE 205 02 Malmö, Sweden

关键词: Exocytosis;    Beta-cell;    Islet of Langerhans;    Insulin secretion;    Diabetes;    Cystic Fibrosis;    CFTR;   

Others  :  854872
DOI  :  10.1186/1741-7015-12-87

 received in 2013-12-09, accepted in 2014-04-10,  发布年份 2014

【 摘 要 】

Background

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene lead to the disease cystic fibrosis (CF). Although patients with CF often have disturbances in glucose metabolism including impaired insulin release, no previous studies have tested the hypothesis that CFTR has a biological function in pancreatic beta-cells.

Methods

Experiments were performed on islets and single beta-cells from human donors and NMRI-mice. Detection of CFTR was investigated using PCR and confocal microscopy. Effects on insulin secretion were measured with radioimmunoassay (RIA). The patch-clamp technique was used to measure ion channel currents and calcium-dependent exocytosis (as changes in membrane capacitance) on single cells with high temporal resolution. Analysis of ultrastructure was done on transmission electron microscopy (TEM) images.

Results

We detected the presence of CFTR and measured a small CFTR conductance in both human and mouse beta-cells. The augmentation of insulin secretion at 16.7 mM glucose by activation of CFTR by cAMP (forskolin (FSK) or GLP-1) was significantly inhibited when CFTR antagonists (GlyH-101 and/or CFTRinh-172) were added. Likewise, capacitance measurements demonstrated reduced cAMP-dependent exocytosis upon CFTR-inhibition, concomitant with a decreased number of docked insulin granules. Finally, our studies demonstrate that CFTR act upstream of the chloride channel Anoctamin 1 (ANO1; TMEM16A) in the regulation of cAMP- and glucose-stimulated insulin secretion.

Conclusion

Our work demonstrates a novel function for CFTR as a regulator of pancreatic beta-cell insulin secretion and exocytosis, and put forward a role for CFTR as regulator of ANO1 and downstream priming of insulin granules prior to fusion and release of insulin. The pronounced regulatory effect of CFTR on insulin secretion is consistent with impaired insulin secretion in patients with CF.

【 授权许可】

   
2014 Edlund et al.; licensee BioMed Central Ltd.

【 图 表 】

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