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ヒト側頭骨病理標本からレーザーキャプチャー・ダイセクションによる難聴遺伝子の抽出

Research Project

Project/Area Number 16659462
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field Otorhinolaryngology
Research InstitutionTokyo Medical and Dental University

Principal Investigator

喜多村 健  東京医科歯科大学, 大学院医歯学総合研究科, 教授 (90010470)

Co-Investigator(Kenkyū-buntansha) 古宇田 寛子  東京医科歯科大学, 大学院医歯学総合研究科, 助手 (80334423)
八島 隆敏  東京医科歯科大学, 医学部附属病院, 助手 (50372438)
Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywords遺伝子 / ゲノム / 細胞・組織 / 神経科学 / 脳・神経 / バイオテクノロジー
Research Abstract

難聴の病態を解析するために、難聴発症の原因が明瞭である難聴遺伝子変異の定量解析と内耳組織病理の相関を検討した。本年度に対象とした症例は、非症候群性遺伝性難聴であるミトコンドリアDNA7511変異である。対象症例のセロイジン包埋側頭骨標本の切片上でレーザーを用いて、蝸牛有毛細胞、血管条、ラセン神経節細胞を数個ずつ採取した。採取した細胞からpure gene kitにてDNAを抽出しTaqMan PCRにて定量的PCRを施行した。当該遺伝子変異は、TaqMan PCRにて増幅可能なプライナー設計が不能と判明した。ミトコンドリアDNA3243変異であるMELAS症例の側頭骨病理においては、遺伝子変異の定量解析と病理変化の相関を既に報告している(Laryngoscope,2003)。本年度は、同じミトコンドリアDNA3243変異であるMELAS症例の側頭骨病理を新たに採取・DNAを抽出し、定量的PCRを施行して、遺伝子変異量と病理変化の相関を解析中である。
ヒト側頭骨を、病理解剖時に採取・凍結保存し、RNA later溶液中にて骨迷路を削り、蝸牛・前庭の膜迷路を採取して、RNAを抽出した。TaqMan probeを用いたりアルタイムPCR法により、COCH、SLC26A4、CDH23、ATP2B2のmRNAの定量を行った。ヒト側頭骨を前期高齢者(65歳以上75歳未満)と超高齢者(90歳以上)の2群に分類して、両者間のmRNAの差を解析中である。

Report

(3 results)
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (16 results)

All 2007 2006 2005 2004 Other

All Journal Article (16 results)

  • [Journal Article] Cross-coupling in a body-translating reaction : interaural optokinetic stimulation reflects a gravitational cue.2007

    • Author(s)
      Tsutsumi T
    • Journal Title

      Acta Oto-Laryngologica 127

      Pages: 273-279

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Vlgrl is required for proper stereocilia maturation of cochlear hair cells.2007

    • Author(s)
      Yagi H
    • Journal Title

      Genes Cells 12

      Pages: 235-250

    • Related Report
      2006 Annual Research Report
  • [Journal Article] SIX1 mutation associated with enlargement of the vestibular aqueduct in a patient with Branchio-Oto syndrome.2006

    • Author(s)
      Ito T
    • Journal Title

      Laryngoscope 116

      Pages: 796-799

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Acute sensorineural hearing loss and vertigo in a young adult with congenital plasminogen disorder.2006

    • Author(s)
      Ishida T
    • Journal Title

      Auris Nasus Larynx 33

      Pages: 187-190

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Multiple quantitative trait loci modify cochlear hair cell degeneration in the Beethoven (Tmcl^<Bth>)mouse model of progressive hearing loss DFNA36.2006

    • Author(s)
      Noguchi Y
    • Journal Title

      Genetics 173

      Pages: 2111-2119

    • Related Report
      2006 Annual Research Report
  • [Journal Article] 総論 遺伝子解析の現状と展望.2006

    • Author(s)
      喜多村 健
    • Journal Title

      JOHNS 22

      Pages: 1677-1680

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Detection of mitochondrial DNA from human inner ear using real-time polymerase chain reaction and laser microdissection.2005

    • Author(s)
      Kimura Y, Kouda H, Eishi Y, Kobayashi D Suzuki Y, Ishige I, Iino Y, Kitamura K
    • Journal Title

      Acta Oto-laryngol 125

      Pages: 697-701

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A mutation in Wolfram syndrome type 1 gene in a Japanese family with autosomal dominant low-frequency sensorineural hearing loss.2005

    • Author(s)
      Noguchi Y, Yashima T, Hatanaka A, Uzawa M, Yasunami M, Kimura A, Kitamura K
    • Journal Title

      Acta Otolaryngol 125

      Pages: 1189-1194

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A clinical, genetic, and neuropathologic study in a family with 16q-linked ADCA type III.2005

    • Author(s)
      Owada K, Ishikawa K, Toru S, Ishida G, Gomyoda M, Tao O, Noguchi Y, Kitamura K, Kondo I, Noguchi E, Arinami T, Mizusawa H
    • Journal Title

      Neurology 65

      Pages: 629-632

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Detection of mitochondrial DNA from human inner ear using real-time polymerase chain reaction and laser microdissection.2005

    • Author(s)
      Kimura Y, Kouda H, Eishi Y, Kobayashi D, Suzuki Y, Ishige I, Iino Y, Kitamura K
    • Journal Title

      Acta Otolaryngologica 125

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Six1 controls patterning of the mouse otic vesicle.2004

    • Author(s)
      Ozaki H, Nakamura K, et al.
    • Journal Title

      Development 131

      Pages: 551-562

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A clinical and genetic study in a large cohort of patients with spinocerebellar ataxia type 6.2004

    • Author(s)
      Takahashi H, Ishikawa K, Tsutsumi T, Fujigasaki H, et al.
    • Journal Title

      J Hum Genet 49

      Pages: 256-264

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A comparison of acute low-tone sensorineural hearing loss versus Meniere's disease in electrocochleography.2004

    • Author(s)
      Noguchi Y, Nishida H, Tokano H, Kawashima Y, Kitamura K
    • Journal Title

      Ann Otol Rhinol Laryngol 113

      Pages: 194-199

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Audiovestibular findings in patients with mitochondrial A1555G mutation.2004

    • Author(s)
      Noguchi Y, Yashima T, Ito T, Sumi T, Tsuzuku T, Kitamura K
    • Journal Title

      Laryngoscope 114

      Pages: 344-348

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Molecular analysis of the temporal bone with laser capture microdissection (LCM) and TaqMan PCR.2004

    • Author(s)
      Koda H, Kimura Y, Takahashi K, Iino Y, Kitamura K
    • Journal Title

      Association for Research in Otolaryngolgoy Abstract

      Pages: 145-146

    • Related Report
      2004 Annual Research Report
  • [Journal Article] SIX1 mutation associated with enlargement of the vestibular aqueduct in a patient with Branchio-Oto syndrome.

    • Author(s)
      Ito T, Noguchi Y, Yashima T, Kitamura K
    • Journal Title

      Laryngoscope (in press)

    • Related Report
      2005 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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