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Journal of Histochemistry and Cytochemistry, Vol. 49, 463-474, April 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Immunolocalization of Anion Exchanger AE2, Na+/H+ Exchangers NHE1 and NHE4, and Vacuolar Type H+-ATPase in Rat Pancreas

Eleni Roussaa,c, Seth L. Alperd, and Frank Thévenodb,c
a Departments of Anatomy, Medical Faculty, University of Saarland, Homburg/Saar, Germany
b Physiology, Medical Faculty, University of Saarland, Homburg/Saar, Germany
c School of Biological Sciences, University of Manchester, Manchester, United Kingdom
d Molecular Medicine and Renal Units, Beth Israel Deaconess Medical Center and Departments of Medicine and Cell Biology, Harvard Medical School, Boston, Massachusetts

Correspondence to: Frank Thévenod, School of Biological Sciences, U. of Manchester, G.38 Stopford Bldg., Oxford Road, Manchester M13 9PT, UK. E-mail: frank.thevenod@man.ac.uk

We have studied the expression and localization of several H+ and HCO3- transporters, whose presence in the rat pancreas is still unclear. The Cl-/HCO3- exchanger AE2, the Na+/H+ exchangers NHE1 and NHE4, and the 31-kD and 70-kD vacuolar H+-ATPase (V-ATPase) subunits were detected by immunoblotting and immunocytochemical techniques. Immunoblotting of plasma membranes with transporter-specific antibodies revealed protein bands at {approx}160 kD for AE2, at {approx}90 kD and {approx}103 kD for NHE1 and NHE4, respectively, and at 31 kD and 70 kD for V-ATPase. NHE1 and NHE4 were further identified by amplification of isoform-specific cDNA using RT-PCR. Immunohistochemistry revealed a basolateral location of AE2, NHE1, and NHE4 in acinar cells. In ducts, NHE1 and NHE4 were basolaterally located but no AE2 expression was detected. V-ATPase was detected in zymogen granules (ZGs) by immunogold labeling, and basolaterally in duct cells by immunohistochemistry. The data indicate that NHE1 and NHE4 are co-expressed in rat pancreatic acini and ducts. Basolateral acinar AE2 could contribute to Cl- uptake and/or pH regulation. V-ATPase may be involved in ZG fusion/exocytosis and ductal HCO3- secretion. The molecular identity of the ductal Cl-/HCO3- exchanger remains unclear.

(J Histochem Cytochem 49:463–474, 2001)

Key Words: bicarbonate transport, secretion, proton pump, exocrine glands


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