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2009 Fiscal Year Annual Research Report

脂肪滴形成へのオートファジーの関与について

Research Project

Project/Area Number 21659050
Research InstitutionJuntendo University

Principal Investigator

内山 安男  Juntendo University, 医学部, 教授 (10049091)

Co-Investigator(Kenkyū-buntansha) 小池 正人  順天堂大学, 医学部, 准教授 (80347210)
佐々木 光穂  順天堂大学, 医学部, 助教 (20432536)
Keywords脂肪滴 / LC3 / オートファジー / 小胞体膜 / エネルギー代謝 / 培養細胞 / 3T3-L1細胞
Research Abstract

私達は、膜型LC3が脂肪滴の表層に局在し、細胞質型から膜型への変換ができないと、脂肪滴の形成が抑制されることを報告した。本年度は、培養がん細胞を用いて脂肪滴の形成と出芽酵母Atg8のほ乳類のオルソログの一つであるLC3との関係を調べた。用いた細胞は、PC12、HeLa、Cos、HepG2、3T3-L1細胞で、これらを70%の密度で培地に播き、12時間後に脂肪染色(BODIPY)すると、たくさんの脂肪滴が形成される。この状態でLC3の染色をすると、BODIPY陽性あるいはperilipin陽性の脂肪滴にLC3が局在すること、脂肪滴分画を採取すると膜型のLC3-IIがperilipinと共に存在することが分かった。そこで、これらの細胞のLC3 mRNAをノックダウンしたところ、LC3の発現が野生型の10から20%に抑制された細胞株を得ることができた。これらの細胞を70%の密度で培養し、12時間後に中性脂肪の量とBODIPY陽性の脂肪滴をを検討した。その結果、どの細胞株も野生型に比べ中性脂肪の量は半減し、BODIPY陽性の脂肪滴の形成は有意に抑制されていた。この現象は、脂肪細胞の前駆細胞である3T3-L1でもみられた。この細胞をサイトカイン(insulin, 3-isobutyl-1-methylxanthine, dexamethasone)存在下で6日間培養すると野生型では、PPAR-γなどの脂肪細胞特異的タンパク質のmRNAが上昇するが、これと共にLC3mRNAも増加することが分かった。この状態では、BODIPY/perilipin陽性の脂肪滴が形成される。しかし、ノックダウン細胞では脂肪滴の形成は有意に抑制されることが明らかとなった。この時、中性脂肪の量も半減していた。これらの結果は、脂肪滴形成に膜型LC3の存在が必要であることを示している。

  • Research Products

    (22 results)

All 2010 2009

All Journal Article (17 results) (of which Peer Reviewed: 15 results) Presentation (5 results)

  • [Journal Article] Atg9A protein, an autophagy-related membrane protein, is localized in the neurons of mouse brains2010

    • Author(s)
      Tamura H, Shibata M, Koike M, Sasaki M, Uchiyama Y
    • Journal Title

      J Histochem Cytochem

      Volume: 58 Pages: 443-453

    • Peer Reviewed
  • [Journal Article] JE Loss of ALS2/Alsin Exacerbates Motor Dysfunction in a SOD1^<H46R>-Expressing Mouse ALS Model by Disturbing Endolysosomal Trafficking2010

    • Author(s)
      Hadano S, Otomo A, Kunita R, Suzuki-Utsunomiya A, Akatsuka A, Koike M, Aoki M, Uchiyama Y, Itoyama Y, Ikeda
    • Journal Title

      PLoSONE (in press)

    • Peer Reviewed
  • [Journal Article] Cathepsin L in bone marrow-derived cells is required for retinal and choroidal neovascularization2010

    • Author(s)
      Shimada N, Ohno-Matsui K, Iseki S, Koike M, Uchiyama Y, Wang J, Yoshida T, Sato T, Peters C, Mochizuki M, Morita I
    • Journal Title

      Am J Pathol (in press)

    • Peer Reviewed
  • [Journal Article] LC3, a microtubule-associated protein1A/B light chain3, is involved in cytoplasmic lipid droplet formation2010

    • Author(s)
      Shibata M, Yoshimura K, Tamura H, Ueno T, Nishimura T, Inoue T, Sasaki M, Koike M, Arai H, Kominami E, Uchiyama Y
    • Journal Title

      Biophys Biochem Res Comm 393

      Pages: 274-279

    • Peer Reviewed
  • [Journal Article] Chondrogenesis during the ES cell differentiation in vitro2010

    • Author(s)
      Yagita Y, Horie K, Yamanaka I, Koinuma S, Shigeyoshi Y, Kawakami K, Takeda J, Uchiyama Y
    • Journal Title

      PNAS 107

      Pages: 3846-3851

    • Peer Reviewed
  • [Journal Article] Nesfatin-1-regulated paraventricular oxytocinergic signaling causes anorexia through a leptin-independent melanocortin pathway2009

    • Author(s)
      Maejima Y, Sedbazar U, Suyama S, Kohno D, Onaka T, Takano E, Yoshida N, Koike M, Uchiyama Y, Fujiwara K, Yashiro T, Horvath TL, Dietrich MO, Tanaka S, Dezaki K, Oh-I S, Hashimoto K, Shimizu H, Nakata M, Mori M, Yada T
    • Journal Title

      Cell Metabolism 10

      Pages: 355-365

    • Peer Reviewed
  • [Journal Article] Ablation of C/EBP beta alleviates ER stress and pancreatic beta cell failure through induction of GRP782009

    • Author(s)
      Matsuda, T., Kido, Y., Nakano-Hashimoto, N., Kaisho, T., Tanaka, T., Asahara, S., Shigeyama, Y., Takeda, A., Inoue, T., Shibutani, Y., Koyanagi, M., Hosooka, T., Matsumoto, M., Inoue, H., Uchida, T., Koike, M., Uchiyama, Y., Akira, S., Kasuga, M.
    • Journal Title

      J Clin Invest 120

      Pages: 115-126

    • Peer Reviewed
  • [Journal Article] Mini Screening of Kinase Inhibitors Affecting Period-length of Mammalian Cellular Circadian Clock2009

    • Author(s)
      Yagita K, Yamanaka I, Koinuma S, Shigeyoshi Y, Uchiyama Y.
    • Journal Title

      Acta Histochem Cytochem 42

      Pages: 89-93

    • Peer Reviewed
  • [Journal Article] Acquisition of T regulatory function in cathepsin L-inhibited T cells by eye-derived CTLA-2alpha during inflammatory conditions2009

    • Author(s)
      Sugita S, Horie S, Nakamura O, Maruyama K, Takase H, Usui Y, Takeuchi M, Ishidoh K, Koike M, Uchiyama Y, Peters C, Yamamoto Y, Mochizuki M.
    • Journal Title

      J Immunol 183

      Pages: 5013-5022

    • Peer Reviewed
  • [Journal Article] Caspase-9 activation revealed by semaphorin 7A cleavage is independent of apoptosis in the aged olfactory bulb2009

    • Author(s)
      Ohsawa S, Hamada S, Asou H, Kuida K, Uchiyama Y, Yoshida H, Miura M.
    • Journal Title

      J Neurosci 29

      Pages: 11385-11292

    • Peer Reviewed
  • [Journal Article] The MAP1-LC3 conjugation system is involved in lipid droplet formation2009

    • Author(s)
      Shibata M*, Yoshimura K*, Furuya M, Koike M, Ueno T, Komatsu M, Arai H, Tanaka K, Kominami E, Uchiyama Y(*equally contributed)
    • Journal Title

      Biophys Biochem Res Comm 382

      Pages: 419-423

    • Peer Reviewed
  • [Journal Article] The novel lipid raft adaptor p18 controls endosome dynamics by anchoring the MEK-ERK pathway to late endosomes2009

    • Author(s)
      Nada S*, Hondo A*, Kasai A*, Koike M*, Saito K, Uchiyama Y, Okada M(*equally contributed)
    • Journal Title

      EMBO J 28

      Pages: 477-489

    • Peer Reviewed
  • [Journal Article] Participation of autophagy in the initiation of graft dysfunction after rat liver transplantation2009

    • Author(s)
      Gotoh K, Lu Z, Morita M, Shibata M, Koike M, Waguri S, DonoK, DokiY, Kominami E, Sugioka A, Monden M, Uchiyama Y
    • Journal Title

      Autophagy 5

      Pages: 351-360

    • Peer Reviewed
  • [Journal Article] Phosphorylation of highly conserved NF-M KSP repeats is not required for myelin-dependent radial axonal growth2009

    • Author(s)
      Garcia M, Rao M, Fujimoto J, Garcia V, Shah S, Crum J, Gotow T, Uchiyama Y, Ellisman MH, Calcutt N, Cleveland D
    • Journal Title

      J Nurosci 29

      Pages: 1277-1284

    • Peer Reviewed
  • [Journal Article] Very-high-dose alpha-tocopherol supplementation increases blood pressure and causes possible adverse central nervous system effects in stroke-prone spontaneously hypertensive rats2009

    • Author(s)
      Miyamoto K, Shiozaki M, Shibata M, Koike M, Uchiyama Y, Gotow T
    • Journal Title

      J Neurosci Res 87

      Pages: 556-566

    • Peer Reviewed
  • [Journal Article]2009

    • Author(s)
      Uchiyama Y, Koike M, Shibata M, Sasaki M
    • Journal Title

      Chapter 3 Autophagic neuron death. In : Autophagy in Disease and Clinical Applications, Part C Vol.453(Edited by : Daniel J.Klionsky)(Elsevier Inc.)

      Pages: 33-51

  • [Journal Article]2009

    • Author(s)
      Uchiyama Y, Koike M, Shibata M
    • Journal Title

      Autophagy and cell death. In : Molecular imaging for integrated medical therapy and drug development(Ed.Yuji Kuge)(Springer Japan)

      Pages: 176-188

  • [Presentation] Autophagy and cell death2009

    • Author(s)
      内山安男
    • Organizer
      5^<th> International Symposium on Autophagy
    • Place of Presentation
      大津プリンスホテル コンベンションホール
    • Year and Date
      20090924-20090928
  • [Presentation] Morphological aspects of autophagy : Identification and possible mechanism of autophagosome formation in neuronal cells2009

    • Author(s)
      内山安男
    • Organizer
      Microscopy Conference 2009
    • Place of Presentation
      オーストリア グラーツ
    • Year and Date
      20090901-20090904
  • [Presentation] Morphological features of programmed cell death across the eukaryotic kingdom2009

    • Author(s)
      内山安男
    • Organizer
      6^<th> ISEM09 Committee
    • Place of Presentation
      神戸大学
    • Year and Date
      2009-09-17
  • [Presentation] Cell Death and Autophagy2009

    • Author(s)
      内山安男
    • Organizer
      International Symposium on Autophagy and Cell Death
    • Place of Presentation
      順天堂大学医学研究科老人性疾患病態・治療研究センター
    • Year and Date
      2009-07-04
  • [Presentation] Autophagy and other pathomechanisms2009

    • Author(s)
      内山安男
    • Organizer
      12^<th> International Congress on Neuronal Ceroid Lipofuscinoses(NCL)
    • Place of Presentation
      ドイツ ハンブルグ
    • Year and Date
      2009-06-05

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Published: 2011-06-16   Modified: 2016-04-21  

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