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Generation of indoleamine 2,3-dioxygenase-dificient mouse

吲哚胺2,3-双加氧酶缺陷型小鼠的产生

基本信息

批准号：
14570098
负责人：
TAKIKAWA Osamu
金额：
$ 2.24万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Indoleamine 2,3-dioxygenase (IDO) is a rate-limiting enzyme in the kynurenine pathway of tryplophan metabolism in various tissues of mammals. IDO is induced dramatically (10-100-fold) during many types of infection or inflammation, resulting in a marked increase in tryptophan metabolism locally or systemically. The IDO-mediated tryptophan metabolism deprives the amino acid almost completely from the cells expressing IDO. Such tryptophan deprivation is shown to be one of the mechanisms by which the host cells inhibit growth of infectious pathogens including viruses and intracellular parasites. The IDO induction, however, produces a large amount of metabolites in the kynurenine pathway of tryptophan metabolism, and some of such metabolites have adverse effects on the cellular function by modification of proteins with them and by then agonistic effects on surface receptors on some cells. For example, the metabolites, kynurenine, 3-hydroxykynurenine, and 3-hydroxykynurenine glucoside, bind covalently to lens proteins and the amount of modified proteins increases with age, probably being responsible in part for a cause of age-related cataract. Another metabolite of tryptophan, quinolinic acid, which has a structural similarity to glutamic acid, is found to be an excitatory neurotoxin through an interaction a glutamic acid (NMDA) receptor in vitro and quinolinic acid thus seems to damage neurons dining infection in the brain. IDO deficient mouse helps us to examine these possible adverse effects in vivo. In this research project, we have constructed a targeting vector for IDO gene with neomycin resistant gene and diphtheria toxin gene. Mouse ES cells transfected with ES cells by electroporation and finally 10 ES clones with disrupted IDO gene were obtained.

吲哚胺2，3-双加氧酶(IDO)是哺乳动物多种组织中酪氨酚代谢的犬尿氨酸途径的限速酶。在许多类型的感染或炎症中，IDO被戏剧性地诱导(10-100倍)，导致局部或全身色氨酸代谢显著增加。IDO介导的色氨酸代谢几乎完全剥夺了表达IDO的细胞中的氨基酸。这种色氨酸缺乏被证明是宿主细胞抑制包括病毒和细胞内寄生虫在内的传染性病原体生长的机制之一。然而，IDO诱导会在色氨酸代谢的犬尿氨酸途径中产生大量代谢物，其中一些代谢物通过修饰蛋白质而对细胞功能产生不利影响，进而对某些细胞上的表面受体产生激动性作用。例如，代谢物犬尿氨酸、3-羟基尿氨酸和3-羟基尿氨酸葡萄糖苷与晶状体蛋白共价结合，修饰蛋白的数量随着年龄的增长而增加，这可能是老年性白内障的部分原因。另一种色氨酸的代谢物，喹啉酸，其结构与谷氨酸相似，在体外被发现是一种兴奋性神经毒素，通过谷氨酸(NMDA)受体与喹啉酸的相互作用，似乎可以损伤大脑中感染的神经元。IDO缺陷小鼠帮助我们在体内检测这些可能的不良影响。在本研究中，我们构建了含有新霉素抗性基因和白喉毒素基因的IDO基因的靶向载体。通过电穿孔方法将ES细胞导入小鼠ES细胞，获得10个IDO基因缺失的ES克隆。