Application of magnetically isolated rat retinal vascular endothelial cells for the determination of transporter gene expression levels at the inner blood–retinal barrier
Application of magnetically isolated rat retinal vascular endothelial cells for the determination of transporter gene expression levels at the inner blood–retinal barrier
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DOI：
10.1111/j.1471-4159.2004.02842.x
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发表时间：
2004-12
影响因子：
4.7
通讯作者：
M. Tomi;K. Hosoya
中科院分区：
文献类型：
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作者：
M. Tomi;K. Hosoya
本研究的目的是使用大鼠视网膜血管内皮细胞（RVEC）的磁分离方法和真实的实时定量PCR分析相结合来定量内血视网膜屏障（BRB）的转运蛋白基因水平。RVEC组分中CD 31、Tie-2、claudin-5、occludin、Jam-1、mdr 1a、oatp 2和oatp 14的转录水平比非RVEC组分中的转录水平高100倍以上，表明这些基因主要在内部BRB表达。RVEC组分中GLUT 1和MCT 1的转录水平在各自的转运蛋白家族中最丰富，表明GLUT 1和MCT 1分别在内部BRB的d-葡萄糖和单羧酸转运中发挥主要作用。总之，磁隔离RVEC的应用能够确定在内部BRB的转运蛋白基因水平，从而增加我们的内部BRB功能的分子水平的理解。
The purpose of the present study was to quantify transporter gene levels at the inner blood–retinal barrier (inner BRB) using a combination of magnetic isolation method for rat retinal vascular endothelial cells (RVEC) and real‐time quantitative PCR analysis. The transcript levels of CD31, Tie‐2, claudin‐5, occludin, Jam‐1, mdr1a, oatp2, and oatp14 in the RVEC fraction were more than 100‐fold greater than those in the non‐RVEC fraction, suggesting that these genes are predominantly expressed at the inner BRB. The transcript levels of GLUT1 and MCT1 in the RVEC fraction were the most abundant in the respective transporter family, suggesting that GLUT1 and MCT1 play a predominant role in d‐glucose and monocarboxylate transport, respectively, at the inner BRB. In conclusion, application of magnetically isolated RVEC is able to determine transporter gene levels at the inner BRB thereby increasing our understanding of inner BRB functions at a molecular level.