The 4F2hc/LAT1 complex transports L-DOPA across the blood-brain barrier

被引:219
作者
Kageyama, T
Nakamura, M
Matsuo, A
Yamasaki, Y
Takakura, Y
Hashida, M
Kanai, Y
Naito, M
Tsuruo, T
Minato, N
Shimohama, S
机构
[1] Kyoto Univ, Grad Sch Med, Dept Neurol, Sakyo Ku, Kyoto 6068507, Japan
[2] Kyoto Univ, Grad Sch Pharmaceut Sci, Dept Drug Delivery Res, Kyoto, Japan
[3] Kyorin Univ, Sch Med, Dept Pharmacol & Toxicol, Tokyo, Japan
[4] Japan Sci & Technol Corp, PRESTO, Tokyo, Japan
[5] Univ Tokyo, Inst Mol & Cellular Biosci, Tokyo, Japan
[6] Kyoto Univ, Grad Sch Med, Dept Immunol & Cell Biol, Kyoto, Japan
关键词
L-DOPA; blood-brain barrier; system L; 4F2hc; LAT1;
D O I
10.1016/S0006-8993(00)02758-X
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
L-DOPA is transported across the blood-brain barrier (BBB) by an amino acid transporter, system L. Recently, it has been demonstrated that system L consists of two subunits, 4F2hc and either LAT1 or LAT2. 4F2hc/LAT1 and 4F2hc/LAT2 show different transport characteristics, while their distribution in the brain has not been determined. To clarify whether 4F2hc/LAT1 participates in L-DOPA transport across the BBB, we first examined the expression of 4F2hc/LAT1 in the mouse brain capillary endothelial cell line, MBEC4, as an in vitro BBB model. Northern hybridization and immunoblotting revealed that both 4F2hc and LAT1 are expressed and form a heterodimer in MBEC4 cells. To confirm whether 4F2hc/LAT1 acts as system L to transport L-DOPA, we characterized L-DOPA uptake into the cells. The uptake process was time-dependent, temperature-sensitive, and Na+-independent. Neutral amino acids with bulky side chains and a bicyclic amino acid, 2-aminobicyclo-[2,2,1]-heptane-2-carboxylic acid (BCH), inhibited L-DOPA uptake into MBEC4 cells to a great extent, while an acidic amino acid, basic amino acids, and glycine had no effect. Other neutral amino acids, such as alanine, asparagine, glutamine, serine, and threonine inhibited L-DOPA uptake by 40-70% at most. These characteristics are more compatible with those of 4F2hc/LAT1, rather than those of 4F2hc/LAT2. Finally, immunohistochemistry;with anti-LAT1 antibody demonstrated that LAT1 is predominantly expressed in the microvessels of the central nervous system. This is the first report showing that the 4F2hc/LAT1 complex participates in L-DOPA transport across the BBB. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
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页码:115 / 121
页数:7
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