Expression of Membrane-bound Carbonic Anhydrases IV, IX, and XIV in the Mouse Heart

Renate J. Scheibe ¹, Gerolf Gros ¹, Seppo Parkkila ¹, Abdul Waheed ¹, Jeffrey H. Grubb ¹, Gul N. Shah ¹, William S. Sly ¹ and Petra Wetzel ^1*

¹ Zentrum Physiologie, Medizinische Hochschule Hannover, Hannover, Germany (RJS,GG, PW); Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, Missouri (AW,JHG,GNS,WSS); and Institute of Medical Technology, University of Tampere and Tampere University Hospital, Tampere, Finland (SP)

^* To whom correspondence should be addressed. E-mail: wetzel.petra{at}mh-hannover.de.

Submitted on April 27, 2006
Accepted on 26 July 2006

	Abstract

The expression of membrane-bound carbonic anhydrases (CAs), of carbonic anhydrase (CA) IV, CA IX, CA XII, and CA XIV, has been investigated in the mouse heart. Western blots using microsomal membranes of wild-type hearts demonstrate a 39 kDa, a 43 kDa and a 54 kDa band representing CA IV, CA IX and CA XIV, respectively, but CA XII could not be detected. The expression of CA IX in the CA IV/CA XIV knockout animals was further confirmed using MOLDI-TOF MS/MS. Cardiac cells were immunostained using anti-CA/FITC and anti--actinin/TRITC as well as anti-CA/FITC and anti-SERCA2/TRITC. The subcellular CA localization was investigated by confocal laser scanning microscopy. The CA localization in the sarcolemmal (SL) membrane was examined by double immunostaining using anti-CA/FITC and anti-MCT-1/TRITC. The CAs showed a distinct distribution pattern in the sarcoplasmic reticulum (SR) membrane. CA XIV is predominantly localized in the longitudinal SR, whereas CA IX is mainly expressed in the terminal SR/t-tubular region. CA IV is present in both SR regions, while CA XII is not found in the SR. In the SL membrane, only CA IV and CA XIV are present. We conclude that CA IV and CA XIV are associated with the SR as well as with the SL membrane, CA IX is located in the terminal SR/t-tubular region, and CA XII is not present in the mouse heart. Therefore, the unique subcellular localization of CA IX and CA XIV in cardiac myocytes suggests different functions of both enzymes in excitation-contraction coupling.

Key Words: carbonic anhydrase, cardiomyocytes, sarcoplasmic reticulum, sarcolemma

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