2000 Fiscal Year Annual Research Report

変異Po cDNA導入培養ラツト末梢神経における髄鞘形成障害

Research Project

Project/Area Number	11670773
Research Institution	School of Health Sciences, Sapporo Medical University
Principal Investigator	舘延忠札幌医科大学, 保健医療学部, 助教授 (80136944)
Co-Investigator(Kenkyū-buntansha)	小塚直樹札幌医科大学, 保健医療学部, 助教授 (90225459) 二宮孝文札幌医科大学, 医学部, 講師 (80156140) 山下利春札幌医科大学, 医学部, 講師 (50167706)
Keywords	変異Po cDNA / ラツト末梢神経 / 遺伝子導入
Research Abstract	今までに、Charcot-Marie-Tooth病症例に新しいPo遺伝子変異であるneucleotide 433A-G、neucleotide 543G-C,neucleotide 603と604間にGGCAの4bpの遺伝子挿入を見いだした。この変異をそれぞれ含むPo cDNAをsite-directed mutagenesis法にて作製した。組み換えアデノウイルスの作製は、最終分化したアデノウイルスベクターを用い、組み換えアデノウイルスの作製はGrahamらの系を用いた。まず野生型と変異型PocDNAを組み換えアデノウイルスの作製のためのベクター(pAd-BglII)ヘサブクローンした。このプラスミドとE1を欠如したアデノウイルスゲノム(pJM17)を293細胞ヘリポフェクタミンを用いてcotransfectionして組み換えアデノウイルスを得た。培養神経細胞の作製は、16日の胎児ラツトの後根神経節をとりだしトリプシンで処理して培養した。最初の2日間は血清を含む培地にて、その後30日は無血清でMEM培地にて培養し、髄鞘形成は坑myelin basic protein抗体を用いてavidin-biotin-peroxidase法にて確認した。現在、野生型と変異型PocDNAを含む組み換えアデノウイルスにtransfectionされた293細胞はx-gal染色にて確認した後に、破壊され,遠沈しその培養上清を上述のラツトの培養神経細胞に感染させている。

Research Products
(6 results)

All Other

All Publications (6 results)

[Publications] Moreira MC,Tachi N,Koenig M: "Homozygosity mapping of Portuguese and Japanese forms of ataxia-oculomotor apraxia (AOA1) to 9p13 and evidence for genetic heterogeneity"American Journal of Human Genetics. (In press). (2001)
[Publications] Tachi N,Kozuka N,Ohya K: "Expression of peripheral myelin protein zero in sural nerve of patients with Charcot-Marie-Tooth disease 1B"Pediatric Neurology. (In press). (2001)
[Publications] Tachi N,Kozuka N,Miyaji M: "A double mutation in a patient with X-linked myotubular myopathy"Pediatric Neurology. (In press). (2001)
[Publications] Tachi N,Kozuka N,Ohya K: "Fukuyama-type congenital muscular dystrophy associated with a lack of C-terminal domain of dystrophin"Pediatric Neurology. (In press). (2001)
[Publications] Tachi N,Kozuka N,Ohya K: "Hereditary cerebellar ataxia with peripheral neuropathy and mental retardation"European Neurology. 43. 82-87 (2000)
[Publications] Tachi N,Ohya K,Chiba S: "Expression of DM locus-associated homeodomain protein in congenital myotonic dystropjy"Journal of Child Neurology. 14. 471-473 (1999)

2000 Fiscal Year Annual Research Report

変異Po cDNA導入培養ラツト末梢神経における髄鞘形成障害

Principal Investigator

舘 延忠 札幌医科大学, 保健医療学部, 助教授 (80136944)

Research Products

[Publications] Moreira MC,Tachi N,Koenig M: "Homozygosity mapping of Portuguese and Japanese forms of ataxia-oculomotor apraxia (AOA1) to 9p13 and evidence for genetic heterogeneity"American Journal of Human Genetics. (In press). (2001)

[Publications] Tachi N,Kozuka N,Ohya K: "Expression of peripheral myelin protein zero in sural nerve of patients with Charcot-Marie-Tooth disease 1B"Pediatric Neurology. (In press). (2001)

[Publications] Tachi N,Kozuka N,Miyaji M: "A double mutation in a patient with X-linked myotubular myopathy"Pediatric Neurology. (In press). (2001)

[Publications] Tachi N,Kozuka N,Ohya K: "Fukuyama-type congenital muscular dystrophy associated with a lack of C-terminal domain of dystrophin"Pediatric Neurology. (In press). (2001)

[Publications] Tachi N,Kozuka N,Ohya K: "Hereditary cerebellar ataxia with peripheral neuropathy and mental retardation"European Neurology. 43. 82-87 (2000)

[Publications] Tachi N,Ohya K,Chiba S: "Expression of DM locus-associated homeodomain protein in congenital myotonic dystropjy"Journal of Child Neurology. 14. 471-473 (1999)

舘延忠札幌医科大学, 保健医療学部, 助教授 (80136944)