Machado-Joseph病患者脳におけるCAGリピートの体細胞モザイクの解析-レーザーキャプチャーマイクロダイセクション法を用いて-

马查多-约瑟夫病患者大脑中 CAG 重复体的体细胞嵌合分析 - 使用激光捕获显微切割 -

• 批准号: 12770322
• 负责人: 嶋崎 晴雄
• 金额: $ 1.28万
• 依托单位: Jichi Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12770322/
• 关键词: Machado-Joseph disease; CAG repeat; somatic mosaicism; laser capture microdissection

Machado-Joseph病患者2例(体細胞のリピート数:患者A;70,患者B;81)の剖検脳より,小脳の凍結切片を作成した.Laser capture micro dissection法により小脳の細胞層(分子層,Purkinje細胞層,顆粒層,白質)毎に細胞群を分離し,DNAを抽出し,PCRを行い,リピート数を決定した.その結果,リピート数は,患者Aの分子層:68.97±0.36(n=33),Purkinje細胞層:68.50±0.49(n=36),顆粒層:69.50±0.23(n=36),白質:70.10±0.37(n=30)であった.分子層・Purkinje細胞層・顆粒層と白質の間で,各々リピート数に有意差を認めた.(Mann-Whitney test, P<0.05)患者Bの分子層:81.04±0.23(n=23),Purkinje細胞層:80.52±0.40(n=27),顆粒層:81.29±0.19(n=38),白質:81.80±0.12(n=25)であった.分子層・Purkinje細胞層・顆粒層と白質の間で,各々リピート数に有意差を認めた.(Mann-Whitney test, P<0.05)今回の実験より,分子層・Purkinje細胞層・顆粒層のリピート数が,白質のリピート数より有意に短いという結果が得られた.これらの結果は,小脳皮質のリピート数が,白質より短いという従来の報告と一致する.同じCAG repeat病であるDRPLAでは,顆粒層が小脳皮質の中では最も短いリピート数を示している.一方Machado-Joseph病では,今回の実験結果より,小脳皮質の層別にはリピート数は有意差がなかったが,顆粒層>分子層>Purkinje細胞層の順にrepeat数が短くなる傾向にあった.

2 cases of Machado-Joseph disease (number of somatic cells: patient A;70; patient B;81) were dissected and frozen sections of small cells were prepared.Laser capture micro dissection method was used to separate small cell layers (molecular layer,Purkinje cell layer, granular layer, straight line). Each cell group was separated,DNA was extracted,PCR was performed, and the number of somatic cells was determined. The results were as follows: molecular layer:68.97±0.36(n=33),Purkinje cell layer:68.50±0.49(n=36), granular layer:69.50±0.23(n= 36), straight line:70.10±0.37(n=30). Molecular layer, Purkinje cell layer, Granular layer and straight line between, each of which is different in number. (Mann-Whitney test, P<0.05) Molecular layer:81.04±0.23(n=23),Purkinje cell layer:80.52±0.40(n=27), Granular layer:81.29±0.19(n=38), Straight line:81.80±0.12(n=25). Molecular layer, Purkinje cell layer, Granular layer and straight line between, each of which is different in number. (Mann-Whitney test, P<0.05) The results of this study were as follows: molecular layer, Purkinje cell layer, granular layer, linear layer, intentionally short middle layer. The result is that the number of cortical cells in the cortex is small, and the number of linear cells in the cortex is small. With CAG repeat disease, DRPLA is the shortest layer in the granular layer, which is located in the middle of the small cortex. Machado-Joseph disease, this time the results of the study, small cortical layer, reverse, reverse,

项目成果

Shimazaki, H., Takiyama, Y., Sakoe, K., Amaike, M.et al.: "Meiotic instability of the CAG repeats in the SCA6/CACNA1A gene in two Japanese SCA6 families"Journal of the Neurological Sciences. 185(2). 101-107 (2001)

Shimazaki, H.、Takiyama, Y.、Sakoe, K.、Amaike, M.等人：“两个日本 SCA6 家族中 SCA6/CACNA1A 基因中 CAG 重复的减数分裂不稳定性”《神经科学杂志》。

Namekawa,M.Takiyama,Y.Sakoe,K.Shimazaki,H. et al.: "A large Japanese SPG4 family with a novel insertion mutation of the SPG4 gene : a clinical and genetic study."Journal of the Neurological Sciences. 185(1). 63-68 (2001)

滑川，M.Takiyama，Y.Sakoe，K.Shimazaki，H.

Namekawa, M., Takiyama, Y., Sakoe, K., Shimazaki, H.et al.: "A large Japanese SPG4 family with a novel insertion mutation of the SPG4 gene : a clinical and genetic study"Journal of the Neurological Sciences. 185(1). 63-68 (2001)

Namekawa, M.、Takiyama, Y.、Sakoe, K.、Shimazaki, H.等人：“具有 SPG4 基因新插入突变的日本 SPG4 大家族：临床和遗传学研究”神经科学杂志

Shimazaki,H.Takiyama,Y.Sakoe,K.Amaike,M. et al.: "Meiotic instability of the CAG repeats in the SCA6/CACNA1A gene in two Japanese SCA6 families."Journal of the Neurological Sciences. (in press). (2001)

岛崎,H.Takiyama,Y.Sakoe,K.Amaike,M.