Identification and cell biological analysis of the gene for benign adult familial myoclonic epilepsy

良性成人家族性肌阵挛癫痫基因的鉴定和细胞生物学分析

• 批准号: 14570954
• 负责人: EBIHARA Mitsuru
• 金额: $ 2.24万
• 依托单位: University of Kyoto (2003)The Institute of Physical and Chemical Research (2002)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570954/
• 关键词: Epilepsy; BAFME; Potassium channel; Channelopathy; Whole genome scan; Linkage analysis; Luciferase assay; channellopathy; Channellopathy; Founder effect

Benign adult familial myoclonic epilepsy (BAFME) is an autosomal dominant idiopathic epilepsy characterized by adult-onset, high penetration rate, myoclonus, tremulous finger movement and infrequent epileptic seizure. BAFME was recognized only in Japan.Linkage analysis was carried out in a Japanese family using ABI Linkage Map Set. Chromosome 2, 7, 8, 10 and 17 were not excluded by linkage analysis, but it was strongly suggested that BAFME susceptibility locus is within 4.4cM on the human chromosome 8q23-24 by using extra markers. We have isolated more than 40 BAC clones covering one third of BAFME susceptibility region. We also had YAC contig spanning this region. By using these BAC and YAC clones, exon trapping was performed, resulted in the isolation of two novel exonic sequences. The isolation of the longest cDNAs clone from cDNA library revealed that they have neither homology with the other gene, nor ORF in their sequences. We also found three different potassium channel genes (KCNQ3, Kv8.1 and Kv9.2) on 8q23-24. Because epilepsies are thought to be channelopathy, these are candidate genes for BAFME. Mutation search was performed on these three potassium channel genes in a Japanese family. We couldn't find any mutation both in the KCNQ3 and Kv8.1 gene, but we found an extra 5' exon and possible alternative 3' end in the Kv8.1 gene. No mutation was found in the Kv9.2 gene either. Analysis of promoter activity was performed by using luciferase assay to determine possible promoter region of these potassium channel genes, resulted in the finding of new essential elements in these promoters. The possibility of founder effect in Japanese families is also excluded by genotyping of the other families with BAFME.

良性成人家族性肌阵挛性癫痫（Benignadulfamilialmyoclonicepilepsy，BAFME）是一种常染色体显性遗传的特发性癫痫，以成人发病、高穿透率、肌阵挛、手指震颤和罕见癫痫发作为特征。利用ABI连锁图谱集对一个日本家系进行了连锁分析。连锁分析未排除2、7、8、10和17号染色体，但利用额外标记，有力地表明BAFME易感位点位于人类染色体8 q23 -24上的4.4cM范围内。我们已经分离出超过40个BAC克隆，覆盖了三分之一的BAFME易感区域。我们也有跨越该区域的YAC重叠群。利用这些BAC和YAC克隆进行外显子捕获，分离出两个新的外显子序列。从cDNA文库中分离出最长的cDNA克隆，发现它们与另一个基因既无同源性，也无ORF。在8 q23 -24上还发现了三个不同的钾通道基因（KCNQ 3、Kv8.1和Kv9.2）。因为癫痫被认为是通道病，这些是BAFME的候选基因。在一个日本家族中对这三个钾通道基因进行突变搜索。在KCNQ 3和Kv8.1基因中均未发现突变，但在Kv8.1基因中发现了一个额外的5'外显子和可能的替代3'末端。Kv9.2基因也未发现突变。通过荧光素酶活性分析确定了这些钾通道基因的可能启动子区域，发现了这些启动子中新的必需元件。通过对其他BAFME家系进行基因分型，也排除了日本家系中创始人效应的可能性。

项目成果

Ebihara, M., Ohba, H., Hattori, E., Yamada, K., Takeo Yoshikawa, T.: "Transcriptional activities of cholecystokinin promoter haplotypes and their relevance to panic disorder susceptibility."Am.J.Med.Genet. Neuropsychiatric Genet. 118B(1). 32-35 (2003)

Ebihara, M.、Ohba, H.、Hattori, E.、Yamada, K.、Takeo Yoshikawa, T.：“胆囊收缩素启动子单倍型的转录活性及其与恐慌症易感性的相关性。”Am.J.Med.Genet。

Itokawa, Masanari, Yamada, Kazuo, Yoshitsugu, Kiyoshi, Toyota, Tomoko, Suga, Toshiro, Ohba, Hisako, Watanabe, Akiko, Hattori, Eiji, Shimizu, Hiromitsu, Kumakura, Tetsuo, Ebihara, Mitsuru, Meerabux, Joanne MA, Toru Michio, Yoshikawa, Takeo: "A microsatelli

糸川、雅成、山田、一夫、吉继、清、丰田、智子、菅、敏郎、大场、久子、渡边、明子、服部、英二、清水、弘光、熊仓、哲夫、海老原、充、Meerabux、Joanne MA、Toru

Hattori E., Yamada K., Ebihara M., Toyota T., Nankai M., Shibuya H., Yoshikawa T.: "Association study of the short tandem repeat in the 5' upstream region of the cholecystokinin gene with mood disorders in the Japanese population"American Journal of Medic

Hattori E.、Yamada K.、Ebihara M.、Toyota T.、Nankai M.、Shibuya H.、Yoshikawa T.：“胆囊收缩素基因 5 上游区域的短串联重复序列与情绪障碍的关联研究

Ebihara, M., Ohba, H., Ohno, S., Yoshikawa, T.: "Genomic organization and promoter analysis of the human nicotinic acetylcholine receptor a6 subunit (CHNRA6) gene : Alu and other elements direct transcriptional repression"Gene. 298. 101-108 (2002)

Ebihara, M.、Ohba, H.、Ohno, S.、Yoshikawa, T.：“人烟碱乙酰胆碱受体 a6 亚基 (CHNRA6) 基因的基因组组织和启动子分析：Alu 和其他元件直接转录抑制”基因。

海老原充, 大羽尚子, Joanne Meerabux, 大久保善朗, 加藤昌明, 豊田倫子, 数藤由美子, 山田和男, 吉川武男: "良性成人型家族性ミオクローヌスてんかん原因遺伝子の探索"てんかん治療研究振興財団研究年報. 15巻. 57-64 (2003)

Mitsuru Ebihara、Naoko Ohba、Joanne Meerabux、Yoshiro Okubo、Masaaki Kato、Rinko Toyota、Yumiko Kazuto、Kazuo Yamada、Takeo Yoshikawa：“寻找导致良性成人家族性肌阵挛癫痫的基因”癫痫治疗研究基金会年度报告 15。卷 57-64 (2003)