Hypoxia directly increases serotonin transport by porcine pulmonary artery endothelial cell plasma membrane vesicles.
To determine whether hypoxia has a direct effect on the plasma membrane transport of serotonin (5-HT), we measured 5-HT transport activity: (1) in plasma membrane vesicles isolated from normoxic and hypoxic endothelial cells, (2) in endothelial cell plasma membrane vesicles that were exposed directly to normoxia or hypoxia, and (3) in endothelial cell monolayers incubated in the presence of 1 x 10(-7) M cycloheximide and exposed to normoxia or hypoxia. A 24-h exposure of endothelial cells to hypoxia resulted in a 40% increase (P less than 0.005) in specific 5-HT transport by plasma membrane vesicles derived from these cells. When plasma membrane vesicles were isolated and then directly exposed to normoxia or hypoxia for 1 h at 37 degrees C, a 31% increase (P less than 0.005) in specific 5-HT transport was observed in hypoxic vesicles. Hypoxia did not alter the Km of 5-HT transport (normoxia = 3.47 microM versus hypoxia = 3.76 microM) but markedly increased the maximal rate of transport (Vmax) (normoxia = 202.4 pmol/min/mg protein versus hypoxia = 317.9 pmol/min/mg protein). Cycloheximide alone had no effect on 5-HT transport by normoxic endothelial cells but did block hypoxia-induced increases in 5-HT uptake in endothelial cell monolayers exposed to 24-h hypoxia. These results indicate that hypoxia increases 5-HT transport in pulmonary artery endothelial cells by a direct effect on the plasma membrane, leading to an increase in the effective number of transporter molecules without alteration in transporter affinity for 5-HT, and possibly by an indirect effect involving de novo protein synthesis.
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Research Service, Veterans Affairs Medical Center, Gainesville, FL 32602.
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D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D003513:Cycloheximide / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism*; D000860:Hypoxia / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D011651:Pulmonary Artery / Q000378:metabolism*; D012701:Serotonin / Q000378:metabolism*; D013552:Swine
D012701:Serotonin; D003513:Cycloheximide