MOLEC GENETIC & BIOCHEM INVESTIGATION OF THIAMINE TRANS

莫莱克基因公司

• 批准号: 2894272
• 负责人: CHARLES K SINGLETON
• 金额: $ 18.54万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2894272
• 关键词: RNase protection assay; antibody formation; brain metabolism; complementary DNA; ethanol; immunofluorescence technique; in situ hybridization; inbreeding; intracellular transport; laboratory rat; membrane transport proteins; molecular cloning; nutrient drug interaction; other phosphotransferase; thiamine; thiamine deficiency; thiamine pyrophosphate; vitamin metabolism

DESCRIPTION: Thiamine deficiency [TD], a frequent complication of alcoholism, contributes significantly to the development of damage in various organ systems, including the brain, of alcoholics and malnourished individuals. Previous work by a number of investigations has focused on reductions in the activities of thiamine-utilizing enzymes as being involved mechanistically and sensitivity to TD. The understanding of the molecular events leading to TD-induced brain damage and to variable tissue- and individual-sensitivity cannot ignore the dietary and intracellular availability of thiamine. The interrelationship between and the relative contribution of alcohol and TD to neural damage remains to be clarified. Our recent work has indicated a complex, cell-type specific regulation of intracellular pools of thiamine and its phosphorylated derivatives, had led to the cloning of the yeast thiamine transporter, and has indicated some of the consequence of having a defective thiamine uptake system. Thiamine uptake is defined here as the transport of thiamine uptake system. Thiamine uptake is defined here as the transport of thiamine into cells and intracellular compartments and its conversion to thiamine pyrophosphate [TPP]. We would like to obtain a better understanding of thiamine uptake and of the interplay between uptake and alcohol consumption. Additionally, we are interested in how thiamine uptake and the concomitant alterations of uptake due to alcohol exposure contribute to tissue-specific and inter individual differences in sensitivity to TD. Thus, we propose a molecular genetic and biochemical study of thiamine uptake in rats exposed to acute and chronic alcohol, to TD, and to a combination of both rat conditions. We will clone and sequence cDNAs for the rat thiamine transporter and thiamine pyrophosphokinase [TPK]. Antibodies to the encoded proteins will be obtained. The clones and the antibodies will be used to examine the qualitative and quantitative tissue distribution and expression of the transporter and TPK in the exposed rats. The effects of alcohol on the biochemical properties of the transporter will be examined. These experiments will be done on outbred and inbred rat strains, the latter of which are differentially susceptible to TD or show different alcohol drinking preferences.

描述：硫胺素缺乏症[TD]，一种常见的并发症， 酒精中毒，大大有助于损害的发展， 酗酒者和营养不良者的各种器官系统，包括大脑 个体以前的一些调查工作集中在 降低硫胺素利用酶的活性， 涉及机械性和对TD的敏感性。的理解 分子事件导致TD诱导的脑损伤， 组织和个人的敏感性不能忽视饮食和 硫胺素的细胞内利用率。之间的相互关系， 酒精和TD对神经损伤的相对作用仍有待进一步研究。 明确了我们最近的研究表明， 细胞内硫胺素库及其磷酸化的调节 衍生物，导致克隆酵母硫胺素转运蛋白， 表明了硫胺素缺陷的一些后果 摄取系统硫胺素摄取在此定义为 硫胺素摄取系统硫胺素摄取在此定义为转运 硫胺素进入细胞和细胞内室及其转化 焦磷酸硫胺素[TPP]。我们希望获得更好的 了解硫胺素的吸收和吸收与 酒精消费。此外，我们感兴趣的是硫胺素 摄取以及由于酒精暴露引起的摄取的伴随变化 有助于组织特异性和个体间差异， 对TD的敏感性因此，我们提出了一种分子遗传学和生物化学的方法， 急性和慢性酒精暴露大鼠的硫胺素摄取研究， TD，以及两种大鼠条件的组合。我们将克隆， 大鼠硫胺素转运蛋白和硫胺素的cDNA序列 焦磷酸激酶[TPK]。编码蛋白的抗体将被 得到了克隆和抗体将用于检查 定性和定量的组织分布和表达， 转运蛋白和TPK。酒精对大脑的影响 将检查转运蛋白的生物化学性质。这些 实验将在远交和近交系大鼠品系上进行，后者是 它们对TD的敏感性不同， 饮酒偏好