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Am J Physiol Gastrointest Liver Physiol (September 4, 2003). doi:10.1152/ajpgi.00158.2003
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Submitted on April 4, 2003
Accepted on August 30, 2003

Cl-HCO3- Exchange is Azetazolamide-sensitive and Activated by a Muscarinic Receptor-induced [Ca2+]i Increase in Salivary Acinar Cells

Ha-Van Nguyen1, Alan Stuart-Tilley2, Seth L. Alper3, and James E. Melvin1*

1 Center for Oral Biology in the Aab Institute of Biomedical Sciences, University of Rochester Medical Center, Rochester, NY, USA
2 Molecular Medicine and Renal Units, Beth Israel Deaconess Medical Center, Boston, MA, USA
3 Department of Medicine, Harvard Medical School, Boston, MA, USA

* To whom correspondence should be addressed. E-mail: james_melvin{at}urmc.rochester.edu.

Large volumes of saliva are generated by transepithelial Cl-movement during parasympathetic, muscarinic receptor stimulation. To gain further insight into a major Cl- uptake mechanism involved in this process we have characterized the anion exchanger (AE) activity in mouse serous parotid and mucous sublingual salivary gland acinar cells. The AE activity in acinar cells was Na+-independent, electroneutral and sensitive to the anion exchange inhibitor DIDS, properties consistent with the AE members of the SLC4A gene family. Localization studies using a specific antibody to the ubiquitously expressed AE2 isoform labeled acini in both parotid and sublingual glands. Western blot analysis detected an approximately 170 kDa protein that was more highly expressed in the plasma membranes of sublingual than in parotid glands. Correspondingly, the DIDS-sensitive Cl-/HCO3- exchanger activity was significantly greater in sublingual acinar cells. The carbonic anhydrase antagonist acetazolamide markedly inhibited, while muscarinic receptor stimulation enhanced the Cl-/HCO3- exchanger activity in acinar cells from both glands. Intracellular Ca2+ chelation prevented muscarinic receptor-induced up-regulation of the anion exchanger, whereas raising the intracellular [Ca2+] with the Ca2+-ATPase inhibitor thapsigargin mimicked the effects of muscarinic receptor stimulation. In summary, carbonic anhydrase activity was essential for regulating Cl-/HCO3- exchange in salivary gland acinar cells. Moreover, muscarinic receptor stimulation enhanced anion exchanger activity through a Ca2+-dependent mechanism. Such forms of regulation may play important roles in modulating fluid and electrolyte secretion by salivary gland acinar cells.




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