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大鼠血脑屏障阳离子氨基酸转运体(y+系统)的鉴定

Identification of the cationic amino acid transporter (System y+) of the rat blood-brain barrier.

作者信息

Stoll J, Wadhwani K C, Smith Q R

机构信息

Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, Bethesda, Maryland 20892.

出版信息

J Neurochem. 1993 May;60(5):1956-9. doi: 10.1111/j.1471-4159.1993.tb13428.x.

Abstract

Cationic amino acids are transported from blood into brain by a saturable carrier at the blood-brain barrier (BBB). The transport properties of this carrier were examined in the rat using an in situ brain perfusion technique. Influx into brain via this system was found to be sodium independent and followed Michaelis-Menten kinetics with half-saturation constants (Km) of 50-100 microM and maximal transport rates of 22-26 nmol/min/g for L-lysine, L-arginine, and L-ornithine. The kinetic properties matched that of System y+, the sodium-independent cationic amino acid transporter, the cDNA for which has been cloned from the mouse. To determine if the cloned receptor is expressed at the BBB, we assayed RNA from rat cerebral microvessels and choroid plexus for the presence of the cloned transporter mRNA by RNase protection. The mRNA was present in both cerebral microvessels and choroid plexus and was enriched in microvessels 38-fold as compared with whole brain. The results indicate that System y+ is present at the BBB and that its mRNA is more densely expressed at cerebral microvessels than in whole brain.

摘要

阳离子氨基酸通过血脑屏障(BBB)上一种可饱和的载体从血液转运至脑内。利用原位脑灌注技术在大鼠身上研究了该载体的转运特性。发现通过该系统进入脑内的过程不依赖于钠,并且遵循米氏动力学,L-赖氨酸、L-精氨酸和L-鸟氨酸的半饱和常数(Km)为50 - 100微摩尔,最大转运速率为22 - 26纳摩尔/分钟/克。其动力学特性与y+系统(不依赖钠的阳离子氨基酸转运体)相匹配,该转运体的cDNA已从小鼠中克隆出来。为了确定克隆的受体是否在血脑屏障处表达,我们通过核糖核酸酶保护法检测大鼠脑微血管和脉络丛的RNA中是否存在克隆的转运体mRNA。该mRNA在脑微血管和脉络丛中均存在,与全脑相比,在微血管中富集了38倍。结果表明,y+系统存在于血脑屏障处,并且其mRNA在脑微血管中的表达比在全脑中更为密集。

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