Endothelial adenosine transporter characterization in perfused guinea pig hearts.
Adenosine (Ado), a smooth muscle vasodilator and modulator of cardiac function, is taken up by many cell types via a saturable transporter, blockable by dipyridamole. To quantitate the influences of endothelial cells in governing the blood-tissue exchange of Ado and its concentration in the interstitial fluid, one must define the permeability-surface area products (PS) for Ado via passive transport through interendothelial gaps [PS(g)(Ado)] and across the endothelial cell luminal membrane (PS(ecl)) in their normal in vivo setting. With the use of the multiple-indicator dilution (MID) technique in Krebs-Ringer perfused, isolated guinea pig hearts (preserving endothelial myocyte geometry) and by separating Ado metabolites by HPLC, we found permeability-surface area products for an extracellular solute, sucrose, via passive transport through interendothelial gaps [PS(g)(Suc)] to be 1.9 +/- 0.6 ml. g(-1). min(-1) (n = 16 MID curves in 4 hearts) and took PS(g)(Ado) to be 1. 2 times PS(g)(Suc). MID curves were obtained with background nontracer Ado concentrations up to 800 micrometer, partially saturating the transporter and reducing its effective PS(ecl) for Ado. The estimated maximum value for PS(ecl) in the absence of background adenosine was 1.1 +/- 0.1 ml. g(-1). min(-1) [maximum rate of transporter conformational change to move the substrate from one side of the membrane to the other (maximal velocity; V(max)) times surface area of 125 +/- 11 nmol. g(-1). min(-1)], and the Michaelis-Menten constant (K(m)) was 114 +/- 12 microM, where +/- indicates 95% confidence limits. Physiologically, only high Ado release with hypoxia or ischemia will partially saturate the transporter.
Schwartz
L M
LM
Department of Bioengineering, University of Washington, Seattle, Washington 98195-7962, USA.
Bukowski
T R
TR
Ploger
J D
JD
Bassingthwaighte
J B
JB
D000241:Adenosine / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D003331:Coronary Vessels / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007201:Indicator Dilution Techniques; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D033703:Nucleoside Transport Proteins; D010477:Perfusion
D002352:Carrier Proteins; D008565:Membrane Proteins; D033703:Nucleoside Transport Proteins; D005947:Glucose; D000241:Adenosine