Functional characterization of rat brain-specific organic anion transporter (Oatp14) at the blood-brain barrier: high affinity transporter for thyroxine.
Oatp14/blood-brain barrier-specific anion transporter 1 (Slc21a14) is a novel member of the organic anion transporting polypeptide (Oatp/OATP) family. Northern blot analysis revealed predominant expression of Oatp14 in the brain, and Western blot analysis revealed its expression in the brain capillary and choroid plexus. Immunohistochemical staining indicated that Oatp14 is expressed in the border of the brain capillary endothelial cells. When expressed in human embryonic kidney 293 cells, Oatp14 transports thyroxine (T4; prothyroid hormone) (Km = 0.18 mum), as well as amphipathic organic anions such as 17beta estradiol-d-17beta-glucuronide (Km = 10 mum), cerivastatin (Km = 1.3 mum), and troglitazone sulfate (Km = 0.76 mum). The uptake of triiodothyronine (T3), an active form produced from T4, was significantly greater in Oatp14-expressed cells than in vector-transfected cells, but the transport activity for T3 was approximately 6-fold lower that for T4. The efflux of T4, preloaded into the cells, from Oatp14-expressed cells was more rapid than that from vector-transfected cells (0.032 versus 0.006 min-1). Therefore, Oatp14 can mediate a bidirectional transport of T4. Sulfobromophthalein, taurocholate, and estrone sulfate were potent inhibitors for Oatp14, whereas digoxin, p-aminohippurate, or leukotriene C4, or organic cations such as tetraetheylammonium or cimetidine had no effect. The expression levels of Oatp14 mRNA and protein were up- and down-regulated under hypo- and hyperthyroid conditions, respectively. Therefore, it may be speculated that Oatp14 plays a role in maintaining the concentration of T4 and, ultimately, T3 in the brain by transporting T4 from the circulating blood to the brain.
Sugiyama
Daisuke
D
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. sugiyama@mol.f.u-tokyo.ac.jp
Kusuhara
Hiroyuki
H
Taniguchi
Hirokazu
H
Ishikawa
Shumpei
S
Nozaki
Yoshitane
Y
Aburatani
Hiroyuki
H
Sugiyama
Yuichi
Y
D000818:Animals; D000838:Anions; D001692:Biological Transport; D001812:Blood-Brain Barrier; D015152:Blotting, Northern; D015153:Blotting, Western; D001921:Brain / Q000196:embryology / Q000378:metabolism*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry*; D002412:Cations; D002460:Cell Line; D002490:Central Nervous System / Q000196:embryology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D015536:Down-Regulation; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D008297:Male; D027361:Organic Anion Transporters / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D027701:Organic Cation Transport Proteins; D011485:Protein Binding; D011725:Pyridines / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D013974:Thyroxine / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D014284:Triiodothyronine / Q000378:metabolism
D000838:Anions; D002352:Carrier Proteins; D002412:Cations; D018076:DNA, Complementary; D027361:Organic Anion Transporters; D027701:Organic Cation Transport Proteins; D011725:Pyridines; C451262:Slco1c1 protein, rat; D014284:Triiodothyronine; C086276:cerivastatin; D013974:Thyroxine