DEVELOPMENT AND ANALYSIS OF A MODEL MOUSE FOR NONKETOTIC HYPERGLYCINEMIA

非酮症高甘氨酸血症模型小鼠的研制与分析

• 批准号: 08672593
• 负责人: KURE Shigeo
• 金额: $ 1.34万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672593/
• 关键词: glycine metabolism; ES cells; transgenic mouse; chromosomal location; CA repeat marker; 非ケトーシス型高グリシン血症; ノックアウトマウス; NMDA受容体; in sity hybridization; 相同組み換え

Nonketotic hyperglycinemia (NKH) is an inherited metabolic disease, characterized by marked elevation of the glycine level in body fluid and severe neurological abnormalities. NKH is caused by defect in the mitochondrial glycine cleavage system (GCS). The GCS consists of the four protein components referred to as P-, T-, H-, and L-proteins. In the central nervus system (CNS) the overall activity of the GCS was high in forebrain and cerebellum, whereas it was hardly detected in spinal cord and brain stem. In line with this observation the glycine content was high in spinal cord and cerebrum, but low in cerebral cortex and cerebellum. To elucidate the molecular pathogenesis of NKH we examines the GCS in mouse brain by using a transgenic technique. We mapped mouse P-protein gene was on chromosome 19C and isolated a intragenic CA repeat marker. We generated a transgenic (Tg) mouse overxpressing the human P-protein cDNA which was under control of potent CAG promoter. The Tg mouse expressed a large amount of human P-protein mRNA and the overall GCS activities in various regions of mouse brain. The glycine content in the spinal cord of the Tg mouse was markedly reduced compared with wild type mouse. These data suggested that the overall activities of the GCS in each region of CNS was closely correlated with the expression level of the P-protein, and the GCS plays a pivotal role in regulation of the glycine content in various regions of CNS.

非酮症高甘氨酸血症（NKH）是一种遗传性代谢疾病，其特征是体液中甘氨酸水平显著升高和严重的神经系统异常。NKH是由线粒体甘氨酸裂解系统（GCS）缺陷引起的。GCS由称为P-、T-、H-和L-蛋白的四种蛋白质组分组成。在中枢神经系统（CNS）中，GCS的总体活性在前脑和小脑中较高，而在脊髓和脑干中几乎检测不到。与此观察结果一致，脊髓和大脑中的甘氨酸含量较高，但大脑皮层和小脑中的甘氨酸含量较低。为了阐明NKH的分子发病机制，我们使用转基因技术检测了小鼠脑中的GCS。将小鼠P蛋白基因定位于19 C染色体上，并分离出一个基因内CA重复序列标记。我们产生了一个转基因（Tg）小鼠过表达的人P-蛋白的cDNA是在有效的CAG启动子的控制下。Tg小鼠表达大量的人P蛋白mRNA和整体GCS活动在小鼠脑的各个区域。与野生型小鼠相比，Tg小鼠脊髓中的甘氨酸含量显著降低。这些数据表明，GCS在中枢神经系统的各个区域的整体活动与P-蛋白的表达水平密切相关，GCS在中枢神经系统的各个区域的甘氨酸含量的调节中起着关键作用。

项目成果

Kure, S.et al.: "A missense mutation(His42Arg)in the T-protein gene from a large Israeli-Arab kindred with nonketotic hyperglycinemia." Human Genetics. in press (1998)

Kure, S.等人：“T 蛋白基因中存在错义突变（His42Arg），来自患有非酮症高血糖症的大型以色列-阿拉伯亲属。”

Takeuchi, K, Kure, S. et al.: "Association of a mutation in thiazide-sensitive Na-Cl cotransporter with familial Gitelmen's syndrome." J.Clin.Endocrin.Metab.81. 4496-4499 (1996)

Takeuchi, K, Kure, S. 等人：“噻嗪类敏感的 Na-Cl 协同转运蛋白突变与家族性 Gitelmen 综合征的关联。”

Kure, S. et al.: "Nonketotic hyperglycinemia:Biochemical,molecular,and neurological aspect." Jpn.J.Hum.Genet.42. 13-22 (1997)

Kure, S. 等人：“非酮症高甘氨酸血症：生化、分子和神经学方面。”

Kure,S.et al.: "Mutation and polymorphic marker analtes of 65k-and 67k-glutamate decarboxylase genes in two families with pyridoxine-dependent epilepsy" Journal of Human Genetics. (in press). (1998)

Kure,S.et al.：“两个吡哆醇依赖性癫痫家族中 65k-和 67k-谷氨酸脱羧酶基因的突变和多态性标记分析”人类遗传学杂志。

Kure,S.et al.: "A subtype of pyridoxine dependent epilepsy with normal CSF glutamate concentration." J.Inher.Metabol.Dis.(in press). (1998)

Kure,S.et al.：“吡哆醇依赖性癫痫的一种亚型，脑脊液谷氨酸浓度正常。”