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Journal of Histochemistry and Cytochemistry, Vol. 49, 1045-1054, August 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Carbonic Anhydrase II Associated with Plasma Membrane in a Human Pancreatic Duct Cell Line (CAPAN-1)

Laetitia Alvarezb, Marjorie Fanjulb, Nicholas Carterc, and Etienne Hollandeb
a Laboratoire de Biologie Cellulaire et Moléculaire des Epithéliums
b Université Paul Sabatier, Toulouse, France
c Department of Child Health, St George's Hospital, Medical School, University of London, London, United Kingdom

Correspondence to: Etienne Hollande, Laboratoire de Biologie Cellulaire et Moléculaire des Epithéliums, Université Paul Sabatier, 38 rue des 36 Ponts, 31400 Toulouse, France. E-mail: hollande@lmtg.ups-tlse.fr

The subcellular distribution of carbonic anhydrase II, either throughout the cytosol or in the cytoplasm close to the apical plasma membrane or vesicular compartments, suggests that this enzyme may have different roles in the regulation of pH in intra- or extracellular compartments. To throw more light on the role of pancreatic carbonic anhydrase II, we examined its expression and subcellular distribution in Capan-1 cells. Immunocytochemical analysis by light, confocal, and electron microscopy, as well as immunoblotting of cell homogenates or purified plasma membranes, was performed. A carbonic anhydrase II of 29 kD associated by weak bonds to the inner leaflet of apical plasma membranes of polarized cells was detected. This enzyme was co-localized with markers of Golgi compartments. Moreover, the defect of its targeting to apical plasma membranes in cells treated with brefeldin A was indicative of its transport by the Golgi apparatus. We show here that a carbonic anhydrase II is associated with the inner leaflet of apical plasma membranes and with the cytosolic side of the endomembranes of human cancerous pancreatic duct cells (Capan-1). These observations point to a role for this enzyme in the regulation of intra- and extracellular pH. (J Histochem Cytochem 49:1045–1053, 2001)

Key Words: carbonic anhydrase II, HCO3- secretion, Golgi apparatus, intracellular trafficking, pancreatic duct cells, cell culture


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