Identification and regulation of the gastric H+/K+-ATPase in the rat heart
V Beisvag1, G Falck, J P Loennechen
1Department of Circulation and Medical Imaging, Faculty of Medicine, The Norwegian University of Science and Technology, Trondheim, Norway.
Unlabelled:
Previous reports indicate that H+/K+-adenosine triphosphatase (ATPase) might be expressed in the heart.
Aims:
The objectives of the present study were to explore the presence of H+/K+-ATPase protein and gene expression in the rat heart and to investigate whether the enzyme could contribute to potassium transport across the sarcolemma.
Methods And Results:
We performed reverse transcription-polymerase chain reaction (RT-PCR) on mRNA from myocardium and isolated cardiomyocytes using primers specific for the gastric H+/K+-ATPase alpha-subunit. The PCR products were sequenced and the predicted gastric H+/K+-ATPase sequence was verified. Western blots from myocardium detected a 34-kDa band and a 94-kDa band, indicating the beta-subunit and alpha-subunit of the gastric H+/K+-ATPase, respectively. Immunocytochemistry detected significant immunoreactivity of the beta-subunit in cardiomyocytes. H+/K+-ATPase-dependent potassium transport was assessed by 86Rb+-uptake in isolated cardiomyocytes. Both ouabain and the selective H+/K+-ATPase inhibitor Schering 28080 reduced 86Rb+-uptake at maximum specific inhibition, by 70 and 25%, respectively; the effects were additive. Competitive RT-PCR analysis indicated a significant upregulation of the myocardial H+/K+-ATPase in heart failure after myocardial infarction.
Conclusion:
The gastric isoform of H+/K+-ATPase is expressed in rat cardiac myocytes, both at transcript and protein levels. Functional studies indicate that the enzyme could contribute to potassium and pHi regulation in cardiomyocytes.
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