TRANSPORT IN ISOLATED MEMBRANES OF BLOOD BRAIN BARRIER

血脑屏障隔离膜中的运输

• 批准号: 2416316
• 负责人: RICHARD A HAWKINS
• 金额: $ 15.76万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2416316
• 关键词: aminoacid transport; apical membrane; blood brain barrier; chemical kinetics; glucose transport; glucose transporter; membrane structure; membrane transport proteins; tissue /cell culture; vascular endothelium; vesicle /vacuole

DESCRIPTION (From applicant's abstract) The purpose of this proposal is to determine the characteristics of glucose and amino acid carriers in the constituent membranes of the blood-brain barrier, using a new technique that we have developed. Both the luminal and abluminal membranes of endothelial cells that constitute the blood-brain barrier will be isolated as vesicles, and measurements of glucose and amino acid transport will be made in vitro. The investigators hypothesize that the distribution, ion dependency, and kinetics of these transport proteins in the luminal and abluminal membranes favor facilitated delivery of glucose and essential amino acids to the brain, but prevent accumulation of nonessential amino acids that serve as neurotransmitters. Furthermore, the investigators propose that the disproportionate stimulation of phenylalanine and tryptophan transport across the blood-brain barrier during hyperammonemia is caused by exchange with glutamine, which has accumulated in the extracellular fluid of the brain. Thus, the following specific aims are proposed: 1) Determine the distribution and the transport kinetics of glucose carriers in the luminal and abluminal membranes of endothelial cells comprising the blood-brain barrier. This will be done by probing the respective membranes with cytochalasin B to quantify the number of carriers, and to use specific antibodies to determine their identity. The kinetic characteristics of the transporter will be determined by measuring glucose uptake in both luminal and abluminal vesicles. Membranes will be derived from bovine cerebral microvessels. 2) Determine the distribution and transport kinetics of neutral amino acid carriers in the individual membranes of the blood-brain barrier. The polar arrangement, characteristics, and classification of neutral amino acid transport proteins will be determined in bovine membrane vesicles by kinetic measurements of transport. 3) Determine if trans-stimulation of phenylalanine, tryptophan, and leucine transport by glutamine can account for their enhanced uptake across the blood-brain barrier during hyperammonemia. Transport kinetics of phenylalanine, tryptophan, and leucine will be measured in membrane vesicles that have been pre-loaded with glutamine, to determine if exchange with glutamine enhances their uptake. The exchange mechanism will be characterized.

描述（摘自申请人摘要）

项目成果

Glutamate permeability at the blood-brain barrier in insulinopenic and insulin-resistant rats.

• DOI: 10.1016/j.metabol.2009.07.022
• 发表时间: 2010-02
• 期刊: METABOLISM-CLINICAL AND EXPERIMENTAL
• 影响因子: 9.8
• 作者: Hawkins, Richard A.;Mokashi, Ashwini;DeJoseph, Mary R.;Vina, Juan R.;Fernstrom, John D.
• 通讯作者: Fernstrom, John D.

Penetration of Glutamate into Brain of 7-day-old Rats

谷氨酸对 7 日龄大鼠大脑的渗透

• DOI: 10.1023/b:mebr.0000007102.78696.e3
• 发表时间: 1997
• 期刊: Metabolic Brain Disease
• 影响因子: 3.6
• 作者: J. Viña;M. DeJoseph;P. A. Hawkins;R. Hawkins
• 通讯作者: R. Hawkins

Neutral amino acid transport by the blood-brain barrier. Membrane vesicle studies.

• DOI: 10.1016/s0021-9258(18)35701-6
• 发表时间: 1992-12
• 期刊: The Journal of biological chemistry
• 作者: M. S. D. Pino;R. Hawkins;D. Peterson

Transport studies using membrane vesicles.

使用膜囊泡的运输研究。

• DOI: 10.1385/1-59259-419-0:233
• 发表时间: 2003
• 期刊: Methods in molecular medicine.
• 作者: Peterson,DarrylR;Hawkins,RichardA
• 通讯作者: Hawkins,RichardA