Axonal degeneration of rat culured nerve introduced mutant aprata in cDNA

大鼠培养神经的轴突变性在cDNA中引入突变体aprata

• 批准号: 15591119
• 负责人: TACHI Nobutada
• 金额: $ 2.43万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591119/
• 关键词: aprataxin; 689 ins T c DNA; cultured rat peripherl nerve; axonal degeneration; 変異aprataxin cDFNA導入; 培養ラツト末鞘神経; 軸策変性; aprataxin cDNA; 689insT

Koenig and we identified a responsible gene "aprataxin" for early-onset cerebellar ataxia with hypoalbuminemia(EOCA-HA) in Japan and cerebellar ataxia with ocular apraxia(AOA) in Portugal. The mutation of aprataxin gene in most EOCA-HA patients showed 689 insertion T(689 ins T) homozygously. First, we made a 689 ins T cDNA by site-directed mutagenesis bases on PCR. Second, we made a constructed adeno virus vector contained both wild and 689 ins T aprataxins. Third, we made a cultured rat peripheral nerve using dorsal root of fetal rat. Those cultured nerves were infected by constructed adeno virus vectors, Myelination and axonal sprouting of cultured nerves infected constructed adeno virus vector were observed by immunohistochenistory using anti-Po antibody and electron microscopy. No differences of myelination and axonal changes were observed in cultured nerves infected constructed adeno virus vector containing both a wild and 689 ins T aprataxin gene.Further more, transgenic mouse containing 689 ins T should be obtained. Matured nerve of this mouse should be analyzed.

Koenig和我们鉴定了日本早发性小脑性共济失调伴低白蛋白血症（EOCA-HA）和葡萄牙小脑性共济失调伴眼用不能（AOA）的相关基因“aprataxin”。绝大多数EOCA-HA患者的aprataxin基因突变均为689插入T（689 ins T）。首先，我们通过PCR定点突变的方法获得了一个689 ins的T cDNA。其次，我们构建了一个同时含有野生型和689 ins T aprataxins的腺病毒载体。第三，以胎鼠背根为材料，建立了体外培养的大鼠周围神经。用构建的腺病毒载体感染培养的神经，用抗Po抗体荧光染色和电镜观察感染腺病毒载体的神经的髓鞘形成和轴突出芽。构建的含野生型和689 ins T aprataxin基因的腺病毒载体感染培养神经后，神经髓鞘和轴突的变化无明显差异，应进一步获得含689 ins T的转基因小鼠。应分析这只小鼠的成熟神经。

项目成果

A new mutation of IGHMBP2 gene in spinal muscular atrophy with

IGHMBP2基因新突变与脊髓性肌萎缩症的关系

• 发表时间: 2005
• 期刊: Pediatr Neurol 32
• 作者: Tachi N;Kikuchi S;Kozuka N
• 通讯作者: Kozuka N

Study of correlation between CTG repeat lengths and muscle power in myotonic dystrophy patients

强直性肌营养不良患者CTG重复长度与肌力的相关性研究

• 发表时间: 2005
• 期刊: Sougou Rhiha 33
• 作者: Kikuchi S;Tachi N;Kozuka N et al.
• 通讯作者: Kozuka N et al.

筋緊張性ジストロフィー3塩基反復配列(CTG repeat)と数と筋力との関係

强直性肌营养不良症三核苷酸重复序列（CTG重复序列）及其数量及其与肌肉力量的关系

• 发表时间: 2005
• 期刊: 総合リハ 33
• 作者: 菊池 真;舘 延忠;小塚直樹 他
• 通讯作者: 小塚直樹 他

A new mutation of IGHMBP2 In spinal muscular atrophy with respiratory disgtress

脊髓性肌萎缩症伴呼吸窘迫的IGHMBP2新突变

• 发表时间: 2005
• 期刊: Pediatr Neural 32
• 作者: Tachi N;Kikuchi S;Kozuka N;Nogami A.
• 通讯作者: Nogami A.

A female case of specific epilepsy after encephalitis

女性脑炎后特异性癫痫1例

• 发表时间: 2003
• 期刊: Rinsyou Syouni Igaku 51
• 作者: Suzuki S;Tachi N;Sakurai N et al.
• 通讯作者: Sakurai N et al.