Ross H J, Wright E M
Brain Res. 1984 Mar 12;295(1):155-60. doi: 10.1016/0006-8993(84)90826-6.
Uptakes of neutral L-amino acids into rabbit choroid plexus apical membrane vesicles were studied using L-[14C]amino acids. Apical membrane vesicles were prepared using Ca2+ precipitation and uptakes of 14C-labeled substrates were measured by a rapid mixing and filtration procedure. Na-dependent, concentrative uptake was observed for proline, histidine and methylaminoisobutyric acid (MeAIB). Phenylalanine and D-glucose uptakes showed no significant Na dependence. Proline uptake was a saturable function of the proline concentration with Jmax 3.5 nmol/(mg min) and Kt 0.3 mM. Competition experiments in the presence of Na indicated that proline and MeAIB are mutually competitive. Proline uptake was also inhibited 25% by phenylalanine, but MeAIB uptake was relatively unaffected. Neither proline nor MeAIB transport was significantly inhibited by glycine. We conclude that proline uptake across rabbit choroid plexus apical membrane vesicles in via an Na-dependent pathway which is shared by MeAIB and, to a minor extent, by phenylalanine, but from which glycine is excluded. Histidine uptake was inhibited by glycine, GABA, phenylalanine, proline and MeAIB. This suggests that histidine may utilize a different pathway in addition to that shared with proline and MeAIB. These transporters should play an active role in the regulation of amino acids in the CSF.
使用L-[¹⁴C]氨基酸研究了中性L-氨基酸进入兔脉络丛顶端膜囊泡的摄取情况。通过Ca²⁺沉淀制备顶端膜囊泡,并采用快速混合和过滤程序测量¹⁴C标记底物的摄取量。观察到脯氨酸、组氨酸和甲基氨基异丁酸(MeAIB)存在钠依赖性的浓缩摄取。苯丙氨酸和D-葡萄糖的摄取未显示出明显的钠依赖性。脯氨酸摄取是脯氨酸浓度的饱和函数,Jmax为3.5 nmol/(mg·min),Kt为0.3 mM。在有钠存在的情况下进行的竞争实验表明,脯氨酸和MeAIB相互竞争。脯氨酸摄取也受到苯丙氨酸25%的抑制,但MeAIB摄取相对不受影响。甘氨酸对脯氨酸和MeAIB的转运均无明显抑制作用。我们得出结论,脯氨酸通过钠依赖性途径进入兔脉络丛顶端膜囊泡,该途径为MeAIB所共有,苯丙氨酸在较小程度上也可利用,而甘氨酸则被排除在外。组氨酸摄取受到甘氨酸、γ-氨基丁酸、苯丙氨酸、脯氨酸和MeAIB的抑制。这表明组氨酸除了与脯氨酸和MeAIB共有的途径外,可能还利用了不同的途径。这些转运体在脑脊液中氨基酸的调节中应发挥积极作用。