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マウス個体老化の分子的把握:老化臓器の遺伝子発現レパートリィの作成

Research Project

Project/Area Number 11877021
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionShinshu University

Principal Investigator

樋口 京一  信州大学, 医学部, 教授 (20173156)

Co-Investigator(Kenkyū-buntansha) 森 政之  信州大学, 医学部, 講師 (60273190)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
Keywords遺伝子発現 / マウス / 精巣 / SAGE / cDNA / シークエンス / 老化 / レパートリー / SAGE法 / 塩基配列 / レパートリィ
Research Abstract

高齢マウスとしては最も広く研究に用いられている系統の一つであるBDF1マウスの雄3ヶ月齢および31ヶ月齢と促進老化のモデルマウスであるSAMP1の雄14ヶ月齢マウスを屠殺し精巣のRNAを分離した。老化の分子的把握を目指すため、精巣での遺伝子発現の差異の網羅的比較をSerial Analysis of Gene Expression (SAGE)法とSuppression Subtractive Hybridization (SSH)法で試みた。
より効率良い解析を目指して14塩基のsingle tagを利用する改変SAGEライブラリーを3種の精巣mRNAから作製した。無作為的に数十クローンのプラスミドの塩基配列を読み、ほぼ予測通りのTagの挿入を確認した。今後、各ライブラリーについて約1000クローン(=25,000 Tags)の塩基配列を読み、3つのライブラリーのデータを比較して老化に伴う精巣での遺伝子発現レパートリーの変化の分子的把握を目指す。
またSAMP1およびSAMR1マウスの精巣mRNAより二本鎖cDNAを合成した後、PCR-Select^<TM>cDNA Subtraction Kit (CLONTECH)をもちいてSAMP1-SAMR1およびSAMR1-SAMP1の二種類のサブトラクションライブラリーを作製した。ディファレンシャルスクリーニングによりボジティブクローンを選択し、さらにノザンブロットにより発現の差異を調べた。既知遺伝子3種類と未知遺伝子2種類に発現の違いが見られた。SAMR1において発現が高かったGsg1およびACTは成熟精子細胞において発現していると報告されている遺伝子であり、未知遺伝子の一つ(Tall)も3週齢以降に発現して来る。これら3つの遺伝子の発現は11ヶ月齢のSAMP1でさらに減少しており精巣の加齢との関連が示された。

Report

(2 results)
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Shoji Masato: "Related Articles Senile ocular amyloidosis in SAM and BALB/c strains of mice."Mech Ageing Dev. 120. 87-94 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Wang Jing: "Wild type ApoA-II gene does not rescue Senescence-accelerated mouse (SAMP1) from short life span and accelerated mortality."J Gerontol B Biol Sci. 55A. B432-439 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] ZhanJun Guo: "Genetic analysis of lifespan in hybrid progeny derived from the SAMP1 mouse strain with accelerated senescence."Mechanisms of Ageing & Development. 118. 435-444 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nishikawa Tomofumi: "Tubular aggregates in the skeletal muscle of the senescence-accelerated mouse ; SAM."Mech Ageing Dev. 114. 89-99 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Umezawa Makiko: "Serum lipid concentrations and mean life span are modulated by dietary polyunsaturated fatty acids in the Senescence-accelerated mouse"J Nutr. 130. 221-227 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yazaki Masahide: "Cardiac amyloid in patients with familial amyloid polyneuropathy consists of abundant wild-type transthyretin."Biochem Biophys Res Commun. 274. 702-706. (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kikuchi Takanobu: "Molecular cloning and characterization of a new neuron-specific homologue of rat polypyrimidine tract binding protein."J Biochem (Tokyo). 128. 811-821 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Xing Yanming: "Transmission of mouse senile amyloidosis. 2001"Lab Invest. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mori Masayuki: "Spontaneous loss-of-function mutations of the 8-oxoguanine DNA glycosylase gene in mice and exploration of the possible implication of the gene in senescence."Free Radical Biology & Medicine. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 樋口京一: "老化促進モデルマウス(SAM)"信州医誌. 48. 7-14 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 中村明宏: "老化モデル動物の性差"Geriatric Medicine. 38. 1833-1836 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mizutani Junichi: "Unique mutation in mitochondorial DNA of Senescence-Accelerated Mouse (SAM) strains"J Heredity. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Gilles Grateuau: "Les Amyloidoses"Flammarion Medicine Science. 580 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 永田和宏: "分子生物学・免疫学キーワード辞典(第2版)"医学書院(印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 永田和宏,森正敬,吉田賢右: "分子シャペロンによる細胞機能制御"シュプリンガー・フェアラーク東京(印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Chiba Takuya: "Mouse senile amyloid deposition is suppressed by adenovirus-mediated overexpression of amyloid-resistant apolipoprotein A-II."Am J Pathol 1999. 155・4. 1319-1326 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Fujisawa Hiromi: "Aminoguanidine supplementation delays the onset of senescence in vitro in dermal fibroblast-like cells from senescence-accelerated mice."J Gerontol A Biol Sci Med Sci. 54・7. B276-B282 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mori Masayuki: "Deletion in the beige gene of the beige rat owing to recombination between LlNE1s."Mamm Genome. 10・7. 692-695 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Higuchi Keiichi: "Senescence-accelerated mouse"Methods Enzymol. 309. 674-686 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Xia Chen: "Genetic typing of the senescence-accelerated mouse (SAM) strains with microsatellite markers ."Mamm Genome. 10・3. 235-238 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Shimizu Motoyuk: "ldentification of peak bone mass QTL in a spontaneously osteoporotic mouse strain."Mamm Genome. 10・2. 81-87 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 日本遺伝子治療学会: "遺伝子治療開発研究ハンドブック"エヌ・ティー・エス. 1061 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 折茂 肇: "新老年学(第2版)"東京大学出版会. 1595 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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