MOLECULAR CLONING OF BLOOD/BRAIN BARRIER NUTRIENT TRANSPORTERS

血液/脑屏障营养转运蛋白的分子克隆

• 批准号: 6112291
• 负责人: ERNEST M WRIGHT
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6112291
• 关键词: Xenopus oocyte; aminoacid transport; blood brain barrier; brain metabolism; capillary; cerebrospinal fluid; choroid plexus; gene expression; glucose transport; immunocytochemistry; in situ hybridization; laboratory rabbit; membrane transport proteins; molecular cloning; northern blottings; transport proteins; vascular endothelium; western blottings

The long term goal of this project is to clone, sequence, express and map the distribution of nutrient transporters in the blood-brain and blood- cerebrospinal-fluid barriers. The endothelial cells of the brain capillaries regulate the uptake of nutrients into the brain, and the epithelial cells of the choroid plexus clear the cerebrospinal fluid of many metabolites and drugs. Relatively little is known about the transporters involved in these vital processes. We will clone blood- brain barrier transporters using two different strategies. The first is to determine if the transporters already cloned from other tissues are expressed at the blood-brain-barrier. A candidate clone for brain capillaries is the renal Na+/myo-mositol cotransporter, since isolated brain microvessels transport this important phospholipid precursor, and a candidate for the choroid plexus is the renal Na+/nucleoside cotransporter. Northern and Western blots, in situ hybridization, and immunocytochemical techniques will be used to map the distribution of candidate clones at the blood-brain interface. The second strategy is to use expression cloning techniques to isolate specific brain capillary and choroid plexus transporters, e.g. The L-type amino acid and Na+- dependent proline transporters. The objective is to use the tools of cell and molecular biology to obtain a detailed understanding of nutrient and metabolite transport between the blood and brain.

该项目的长期目标是克隆，序列，表达和映射 营养转运蛋白在血脑和血液中的分布 脑脊液流体屏障。 大脑的内皮细胞 毛细血管调节养分吸收大脑，并调节 脉络丛的上皮细胞清除脑脊液的脑脊液 许多代谢产物和药物。 关于 参与这些重要过程的转运蛋白。 我们将克隆血液 - 脑屏障转运蛋白使用两种不同的策略。 第一个是 确定转运蛋白是否已经从其他组织克隆了 在血脑屏障上表达。 大脑的候选克隆 毛细血管是肾脏Na+/肌醇共转运蛋白，因为 大脑微血管运输这一重要的磷脂前体，并 脉络丛的候选者是肾脏Na+/核苷 共转运者。 北部和韦斯特印迹，原位杂交，以及 免疫细胞化学技术将用于绘制 血脑界面处的候选克隆。 第二个策略是 使用表达克隆技术隔离特定的脑毛细管 和脉络丛转运蛋白，例如L型氨基酸和Na+ - 依赖性脯氨酸转运蛋白。 目的是使用 细胞和分子生物学以获得对养分的详细了解 以及血液和大脑之间的代谢产物转运。