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First published online September 16, 2005
Journal of Experimental Biology 208, 3665-3674 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01815
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Perfusion of the isolated trout heart coronary circulation with red blood cells: effects of oxygen supply and nitrite on coronary flow and myocardial oxygen consumption

F. B. Jensen1,* and C. Agnisola2

1 Institute of Biology, University of Southern Denmark, DK-5230 Odense M, Denmark
2 Department of Biological Sciences, University of Naples Federico II, Napoli, Italy

* Author for correspondence (e-mail: fbj{at}biology.sdu.dk)

Accepted 28 July 2005

A method for perfusion of the isolated trout heart coronary circulation with red blood cells (RBCs) was developed. The method was used to analyse the influence of RBC perfusion on myocardial O2 supply and O2 consumption and to test the hypothesis that nitrite is converted to vasoactive nitric oxide in the RBC-perfused coronary circulation. Perfusion with RBCs significantly increased myocardial O2 supply and O2 consumption by increasing the incoming O2 concentration and the O2 extraction. Coronary flow did not differ between RBC perfusion and saline perfusion, but RBC perfusion established a strong linear increase in myocardial O2 consumption with coronary flow. Nitric oxide was measured in the atrial effluent of the preparation. Perfusion with saline under hypoxic conditions was associated with NO production. The nitric oxide synthase inhibitor L-NA obliterated this NO production and significantly decreased coronary flow, showing that the NO was vasoactive and probably of endothelial origin. RBC perfusion at low PO2 similarly caused an L-NA-inhibitable NO production. The change in NO production upon subsequent nitrite addition, by contrast, was not inhibited by L-NA. Nitrite entered trout erythrocytes independent of degree of oxygenation, but the O2 saturation of RBCs showed a major decrease in the coronary circulation, and [NO2-] decreased while methaemoglobin rose, suggesting that deoxyHb-mediated reduction of nitrite to NO may have occurred. However, other possibilities (e.g. NO2-->NO conversion in myocardial cells) cannot be excluded. The NO formation associated with nitrite had no effect on coronary flow, possibly because NO was produced after the resistance vessels.

Key words: erythrocyte, haemoglobin, microcirculation, nitrite, nitric oxide, vasodilation


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© The Company of Biologists Ltd 2005