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Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada 89557-0046
The molecular identification of
cardiac chloride channels has provided probes to investigate their
distribution and abundance in heart. In this study, the molecular
expression and distribution of volume-regulated chloride channels ClC-2
and ClC-3 in cardiac tissues were analyzed and quantified. Total RNA
was isolated from atria and ventricles of several species (dog, guinea
pig, and rat) and subjected to a quantitative RT-PCR strategy. ClC-2
and ClC-3 mRNA expression were calculated relative to 
-actin
expression within these same tissues. The transcriptional levels of
ClC-3 mRNA were between 1.8 and 10.2% of -actin expression in atria and between 3.4 and 8.6% of -actin in ventricles (n = 3 for each tissue). The levels of ClC-2 in both atria and ventricles
were significantly less than those measured for ClC-3
(n = 3; P < 0.05). ClC-2 mRNA levels
were between 0.04-0.08% and 0.03-0.18% of -actin expression in atria and ventricles, respectively (n = 3 for each tissue). Immunoblots of atrial and ventricular wall protein
extracts demonstrated ClC-2- and ClC-3-specific immunoreactivity at 97 and 85 kDa, respectively. Immunohistochemical localization in guinea
pig cardiac muscle demonstrates a ubiquitous distribution of ClC-2 and
ClC-3 channels in the atrial and ventricular wall. Confocal analysis
detected colocalization of ClC-2 and ClC-3 in sarcolemmal membranes and
distinct ClC-3 immunoreactivity in cytoplasmic regions. The molecular
expression of ClC-2 and ClC-3 in cardiac tissue is consistent with the
proposed role of these chloride channels in the regulation of cardiac
cell volume and the modulation of cardiac electrical activity.
heart; anion channel; immunohistochemistry; cDNA
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