Molecular cytogenetic study of the myotonic dystrophy by in situ hybrdization (ISH)

原位杂交（ISH）对强直性肌营养不良的分子细胞遗传学研究

• 批准号: 09670644
• 负责人: SAITO Fumiko
• 金额: $ 1.98万
• 依托单位: Tokyo Medical & Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670644/
• 关键词: triplet repeat; myotonic dystrophy; ISH method; DM protein kinase; RNA dynamics

Myotonic dystrophy (DM) is an autosomal dominant disease, whose responsible gene encodes DM protein kinase (DMPK) and is located on the chromosome 19q 13. Though DM is caused by abnormal expansion of triplet (CTG) repeat at 3'UTR of DMPK gene, the molecular mechanism of the causing disease have been still unknown.In this study, we have gained the following results by using in situ hybridization (ISH) method to search the molecular pathogenesis of DM.It shows that (1) DMPK transcripts from mutant allele were accumulated in the nuclei of the LCL obtained from the patients, (2) the accumulated transcripts were mainly mRNA rather than pre-mRNA from mutant allele. Moreover, the amount of mRNA in the nuclei of the patients was found to be significantly higher than those in the nuclei of the normal individuals by the molecular biological study, which supports the result by ISH method described above.As the result that the processed mIRNA transcribed from mutant allele are accumulated in the nuclei of the patients may provide a key point to study the pathogenesis of the patients with various clinical features, we will make a further study to resolve the problem.

强直性肌营养不良（DM）是一种常染色体显性遗传疾病，其致病基因编码DM蛋白激酶（DMPK），位于染色体19q 13。虽然DM是由DMPK基因3'UTR处三联体（CTG）重复异常扩增引起的，但其致病的分子机制仍不清楚。在本研究中，我们通过使用 原位杂交（ISH）方法探索DM的分子发病机制。结果显示，（1）突变等位基因的DMPK转录本在患者LCL的细胞核中积累，（2）积累的转录本主要是突变等位基因的mRNA而不是前mRNA。而且，通过分子生物学研究发现，患者细胞核中的mRNA量明显高于正常个体的细胞核中的mRNA量，这支持了上述ISH方法的结果。由于突变等位基因转录的加工后的mRNA在患者细胞核中积累，可能为研究具有各种临床特征的患者的发病机制提供关键点，我们将进一步研究以解决该问题。

项目成果

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Song J：“KNSL4 和 Maz 的人类基因在染色体 16p11.2 上彼此靠近。”

F.Saito-Ohara: "Assignment of the mouse heme oxygenase genes:heme oxygenase-1(Hmox1)to chromosome 16 band B1" Cylogenetics Cell Genetics. 77・1. 180-181 (1997)

F.Saito-Ohara：“小鼠血红素加氧酶基因：血红素加氧酶-1(Hmox1)至染色体16带B1的分配”细胞遗传学77·1(1997)。

X.Tang: "Assignment of the human gene for KBP2/RBP-Jk to chromosome 9p12-13 and 9p13 by fluorescence in situ hybridization" Japanese Journal Human Genetics. 42・2. 337-341 (1997)

X.Tang：“通过荧光原位杂交将 KBP2/RBP-Jk 的人类基因分配到染色体 9p12-13 和 9p13”，日本《人类遗传学》杂志 42·2（1997）。

Orii K O: "Assignment of the gene for very-long-chain acyl-CoA dehydrogenase (Acadv1) to mouse chromosome band 11B2-B5 by in situ hybridization." Cytogenet.Cell Genet.78. 25-26 (1997)

Orii K O：“通过原位杂交将超长链酰基辅酶 A 脱氢酶 (Acadv1) 基因分配至小鼠染色体带 11B2-B5。”

Saito-Ohara F., Ikeuchi T., Matsumoto M.and Kurata S.: "Assignment of the mouse heme oxygenase genes : ^-heme oxygenase-1 (Hmox1) to chromosome 10 band C1 and heme oxygenase-2 (Hmox2)to chromosome 16 band B1." Cytogenet. Cell Genet.77. 180-181 (1997)

Saito-Ohara F.、Ikeuchi T.、Matsumoto M.和 Kurata S.：“小鼠血红素加氧酶基因的分配：^-血红素加氧酶-1 (Hmox1) 至染色体 10 带 C1 和血红素加氧酶-2 (Hmox2) 至