title: Modulation of monocarboxylic acid transporter-1 kinetic function by the cAMP signaling pathway in rat brain endothelial cells.
abstract: MCT1 (monocarboxylic acid transporter 1) facilitates bidirectional monocarboxylic acid transport across membranes. MCT1 function and regulation have not been characterized previously in cerebral endothelial cells but may be important during normal cerebral energy metabolism and during brain diseases such as stroke. Here, by using the cytoplasmic pH indicator 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein-acetoxymethyl ester, the initial rates of monocarboxylate-dependent cytoplasmic acidification were measured as an indication of MCT1 kinetic function in vitro using the rat brain endothelial cell (RBE4) model of blood-brain transport. The initial rate of L-lactate-dependent acidification was significantly inhibited by 5-10-min incubations with agonists of intracellular cAMP-dependent cell signaling pathways as follows: dibutyryl cAMP, forskolin, and isoproterenol. Isoproterenol reduced V(max) but did not affect K(m) values. The effects of forskolin were completely reversed by the protein kinase A inhibitor H89, whereas H89 alone increased transport rates. Cytoplasmic cAMP levels, measured by radioimmunoassay, were increased by forskolin or isoproterenol, and the effect of isoproterenol was inhibited by propranolol. MCT1-independent intracellular pH control mechanisms did not contribute to the forskolin or H89 effects on MCT1 kinetic function as determined with amiloride, monocarboxylate-independent acid loading, or the transport inhibitor alpha-cyano-4-hydroxycinnamate. The data demonstrate the direct modulation of MCT1 kinetic function in cerebral endothelial cells by agents known to affect the beta-adrenergic receptor/adenylyl cyclase/cAMP/protein kinase A intracellular signaling pathway.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Minnesota Medical School, 1035 University Drive, Duluth, MN 55812, USA.
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Smith
authors.forename: Lester R
authors.initials: LR
authors.lastname: Drewes
mesh_terms: D000818:Animals; D001921:Brain / Q000473:pathology*; D002545:Brain Ischemia / Q000473:pathology; D002138:Calibration; D005576:Colforsin / Q000494:pharmacology; D003373:Coumaric Acids / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D007545:Isoproterenol / Q000494:pharmacology; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D027501:Monocarboxylic Acid Transporters / Q000737:chemistry* / Q000378:metabolism; D011863:Radioimmunoassay; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013449:Sulfonamides / Q000494:pharmacology; D027981:Symporters / Q000737:chemistry* / Q000378:metabolism; D000099233:Monocarboxylate Transport Protein 1
chemical_list: D005576:Colforsin; D003373:Coumaric Acids; D000242:Cyclic AMP; D004791:Enzyme Inhibitors; D007545:Isoproterenol; D007546:Isoquinolines; D027501:Monocarboxylic Acid Transporters; D013449:Sulfonamides; D027981:Symporters; D000099233:Monocarboxylate Transport Protein 1; C007175:alpha-cyano-4-hydroxycinnamate; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide