DNA diagnosis of congenital myotonic dystrophy

先天性强直性肌营养不良的DNA诊断

• 批准号: 05670685
• 负责人: TACHI Nobutada
• 金额: $ 1.22万
• 依托单位: Sapporo Mecial University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670685/
• 关键词: Congenital myotonic dystrophy; DNA diagnosis; Myotonic dystrophy protein kinase; meiotic instability; 遺伝子; RT-PCR; MT-PK mRNA; DM Kinase; 合成ポリペプチド; 神経筋接合部; 筋紡錘; 筋細胞膜; 培養筋; 先天性筋緊張性ジストロフィー症; 遺伝子診断; unstable DNA

We have analyzed CTG repeat expansion in 55 myotonic dystrophy (DM) patients, including 25 congenital DM by Southern hybridization using DM gene specific probe (p5B1.4). The CTG repeat was expanded in DM patients when transmitted from parent to child severity. DM families showed genetic anticipation, an increase in disease severity and earlier age of onset in successive generations. The transmission of congenital DM was considered exclusively maternal. We present the first congenital DM patients transmitted from asymptomatic father on the basis of molecular analysis. Based on analysis of CTG repeat in various tissues from a patient of congenital DM,the presence of somatic instability was disclosed.We have analyzed the size of the CTG repeat of DNAs extracted from skeletal muscle and lymphocytes in congenital DM patients. The size from skeletal muscle showed an increase of about 1.5kb to 3.5kb larger than that from lymphocytes in all patients.We present expression of DM-PK on biopsied muscles and cultured muscles by immunocytochemistry using antibody aganist synthetic DM-PK peptide antigen. DM-PK was observed in neuromuscular junctions, muscle spindle, and sarcolemma on biopsied muscles. On cultured muscles, DM-PK was expressed in cytoplasma of myoblast and myotube.

应用DM基因特异性探针（p5B1.4）对55例强直性肌营养不良（DM）患者（包括25例先天性DM）的CTG重复序列扩增进行了Southern杂交分析。当从父母传播到孩子时，CTG重复在DM患者中扩大。DM家系表现出遗传预测、疾病严重程度增加和连续几代发病年龄提前。先天性糖尿病的传播被认为完全是母亲的。我们提出了第一个先天性糖尿病患者传播的无症状的父亲的分子分析的基础上。通过对一例先天性DM患者不同组织中CTG重复序列的分析，揭示了体细胞不稳定性的存在，并分析了先天性DM患者骨骼肌和淋巴细胞DNA中CTG重复序列的大小。所有患者骨骼肌中DM-PK的大小均比淋巴细胞中的大1.5 ~ 3.5kb。在活检肌肉的神经肌肉接头、肌梭和肌膜中观察到DM-PK。在培养的肌肉中，DM-PK在成肌细胞和肌管的胞浆中表达。

项目成果

Tachi N,Watanabe Y,Kozuka N,Ohya K,Chiba S.: "Immunocytochemical localization of myotonic dystrophy protein kinase in cultured muscle." Acta Histochem Cytochem. 28. 37-39 (1995)

Tachi N，Watanabe Y，Kozuka N，Ohya K，Chiba S.：“强直性肌营养不良蛋白激酶在培养肌肉中的免疫细胞化学定位。”

Ohya K,Tachi,N: "Congenital myotonic dystrophy transmitted from an asymptomatic father with myotonic dystrophy specific gene" Neurology. 44. 1958-1960 (1994)

Ohya K，Tachi，N：“先天性强直性肌营养不良症是由具有强直性肌营养不良症特异性基因的无症状父亲传播的”神经病学。

舘 延忠: "今日の小児治療指針" 医学書院, 766 (1993)

Nobutada Tate：《当今的儿科治疗指南》Igakushoin，766 (1993)

Tachi N et al: "Expression of myotonic dystrophy protein kinase in biopsied muscles" J Neurol Sci. (in press).

Tachi N 等人：“强直性肌营养不良蛋白激酶在活检肌肉中的表达”J Neurol Sci。

Tachi N et al: "Unstable DNA in a patient with a severe form of a congenital myotonic dystrophy" J Neurol Sci. 119. 180-182 (1993)

Tachi N 等人：“严重先天性强直性肌营养不良患者的 DNA 不稳定”J Neurol Sci。