Purinergic regulation of CFTR and Ca(2+)-activated Cl(-) channels and K(+) channels in human pancreatic duct epithelium.

Abstract

Purinergic agonists have been considered for the treatment of respiratory epithelia in cystic fibrosis (CF) patients. The pancreas, one of the most seriously affected organs in CF, expresses various purinergic receptors. Studies on the rodent pancreas show that purinergic signaling regulates pancreatic secretion. In the present study we aim to identify Cl(-) and K(+) channels in human pancreatic ducts and their regulation by purinergic receptors. Human pancreatic duct epithelia formed by Capan-1 or CFPAC-1 cells were studied in open-circuit Ussing chambers. In Capan-1 cells, ATP/UTP effects were dependent on intracellular Ca(2+). Apically applied ATP/UTP stimulated CF transmembrane conductance regulator (CFTR) and Ca(2+)-activated Cl(-) (CaCC) channels, which were inhibited by CFTRinh-172 and niflumic acid, respectively. The basolaterally applied ATP stimulated CFTR. In CFPAC-1 cells, which have mutated CFTR, basolateral ATP and UTP had negligible effects. In addition to Cl(-) transport in Capan-1 cells, the effects of 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one (DC-EBIO) and clotrimazole indicated functional expression of the intermediate conductance K(+) channels (IK, KCa3.1). The apical effects of ATP/UTP were greatly potentiated by the IK channel opener DC-EBIO. Determination of RNA and protein levels revealed that Capan-1 cells have high expression of TMEM16A (ANO1), a likely CaCC candidate. We conclude that in human pancreatic duct cells ATP/UTP regulates via purinergic receptors both Cl(-) channels (TMEM16A/ANO1 and CFTR) and K(+) channels (IK). The K(+) channels provide the driving force for Cl(-)-channel-dependent secretion, and luminal ATP provided locally or secreted from acini may potentiate secretory processes. Future strategies in augmenting pancreatic duct function should consider sidedness of purinergic signaling and the essential role of K(+) channels.

• Authors: Jing Wang, Kristian A Haanes, Ivana Novak
• Journal: American journal of physiology. Cell physiology (Am J Physiol Cell Physiol) Vol. 304 Issue 7 Pg. C673-84 (Apr 01 2013) ISSN: 1522-1563 [Electronic] United States
• PMID: 23364268 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Chloride Channels
• Potassium Channels, Calcium-Activated
• Receptors, Purinergic P2X
• Receptors, Purinergic P2Y
• Cystic Fibrosis Transmembrane Conductance Regulator
• Adenosine Triphosphate
• Calcium
• Uridine Triphosphate

Topics

• Adenosine Triphosphate (pharmacology)
• Calcium (metabolism)
• Cell Line
• Chloride Channels (genetics, metabolism)
• Cystic Fibrosis Transmembrane Conductance Regulator (genetics, metabolism)
• Epithelial Cells (drug effects, metabolism)
• Gene Expression Regulation
• Humans
• Pancreatic Ducts (cytology, metabolism)
• Potassium Channels, Calcium-Activated (genetics, metabolism)
• Receptors, Purinergic P2X
• Receptors, Purinergic P2Y
• Uridine Triphosphate (pharmacology)