Coculture of astroglial and vascular endothelial cells as apposing layers enhances the transcellular transport of hypoxanthine.
In brain, astrocytes and endothelial cells are a major site of adenosine degradation. These two cell types, found in close apposition, constitute the wall of the brain's capillaries and serve as a site of hypoxanthine production and degradation. Both cell types possess the hypoxanthine salvage pathway and can incorporate hypoxanthine into nucleotides. This suggests that the endothelial-astrocyte anatomical complex might play an important role in the brain's purine homeostasis. To test this hypothesis, cocultures of monolayers of vascular endothelial cells and astrocytes were grown over a porous membrane, in close apposition to one another, and studies on hypoxanthine transport and metabolism to uric acid were performed. The flux of hypoxanthine across the cell layers was simultaneously determined and compared with the flux of sucrose, as a probe of passive diffusion. Our results show that in endothelial, glial, and endothelial-glial cell layers the hypoxanthine flux was greater than that of sucrose, and that the flux of hypoxanthine, but not of sucrose, was inhibited by adenine or by lowering the temperature. These results suggest that hypoxanthine moves across endothelial, glial, and endothelial-glial cell layers by a transport process. Furthermore, we found that hypoxanthine transport is enhanced when glial and endothelial cells are cocultured compared with that in glial or endothelial monolayers. In addition the coculture also resulted in a depression of xanthine oxidase activity.
Ceballos
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G
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville 22908.
Rubio
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R
D000225:Adenine / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000378:metabolism*; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D009457:Neuroglia / Q000378:metabolism*; D013696:Temperature
D007042:Hypoxanthines; D019271:Hypoxanthine; D000225:Adenine