1997 Fiscal Year Annual Research Report

筋細胞系をモデルとした細胞増殖と分化の制御機構の研究

Research Project

Project/Area Number	08458242
Research Institution	National Institute of Neuroscience, NCNP
Principal Investigator	鍋島陽一国立精神・神経センター, 神経研究所・遺伝子工学, 部長 (60108024)
Co-Investigator(Kenkyū-buntansha)	吉田松生国立精神, 神経センター・阪大・細胞生体工学センター, 助手 (60294138) 武田伸一国立精神, 神経センター・神経研究所・遺伝子工学, 室長 (90171644) 藤沢淳子国立精神, 神経センター・神経研究所・遺伝子工学, 室長 (60209038)
Keywords	筋分化制御因子 / Myogenin / 筋発生 / 遺伝子ノックアウト / 細胞分化 / 増殖因子 / 細胞培養 / bHLH
Research Abstract	マウス筋芽細胞、C2C12は増殖因子存在下では増殖を続けるが、培地より増殖因子を除くと分化を開始する。その際に細胞は筋管細胞へと分化する細胞と分化せずに筋芽細胞のままで留まる“reserve cell"の2つのサブポピュレーションに分れる。myogeninの発現、細胞分裂からの離脱、P21の発現、細胞の融合など、分化の指標と考えられている事柄は前者の分化する細胞のみで観察される。後者のreserve cellは分化せずに留まり、極めて遅い速度で増殖を続けるようになるが、元のの筋芽細胞としての性質を保持しており、再度、増殖因子を添加すると、増殖を開始し、増殖因子を取り除くと、元の細胞と同じように分化する細胞とreserve cellに分かれる。この2つのサブポピュレーションへの仕分けは分化誘導の極めて早い段階でのMyoD(myf-5)の発現によって導き出される。分化する細胞では増殖中の筋芽細胞で発現しているMyoD(myf-5)の発現が維持されるが。reserve cellではその発現が失われる。さらに、MyoDを強制発現させると、reserve cellを分化する細胞に転換させることができる。この事実はMyoDの発現がC2C12細胞の2つのサブポピュレーションへの仕分けを制御しており、細胞分化の過程そのものの中に増殖能を保持した細胞を残す仕組みをもっていることを示唆している。分化誘導に伴いMyoDの発現を制御している因子を解析するために、培養筋芽細胞の運命を継続的にトレースし、細胞周期と分化誘導をかけたタイミングについて検討したが、明瞭な結論には至らなかった。細胞分裂の際に不等分裂をおこして2つのタイプの細胞が生まれる可能性についても否定的な結果をえている。

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] Yoshida S., FujisawaーSehara., Arai K. and Nabeshima Y.: "Lysophosphatidic acid and bFGF control different modes in proliferating myoblasts." J.Cell.Biol.132. 181-193 (1996)
[Publications] Hoshino M., Suzuki E., Nabeshima Y. and Hama C.: "hikaru genki protein secreted into synaptic cleft from an early stage of synapse formation in Drosopiola." Development. 122. 589-597 (1996)
[Publications] Ohbayashi T., Schmidt E.E., Makino Y., Kishimoto T., Nabeshima Y., Muramatsu M. and Tamura T.: "Promotor structure of the mouse TATA-binding protein (TBP)gene." Biochem.Biophys.Res.Comm.225. 257-280 (1996)
[Publications] Sone M., Hoshino M., Suzuki E., Saigo K., Nabeshima Y. and Hama C.: "Still life,a protein in synaptic tereminals of Drosophila homologous to GDP-GTP exchanger." Science. 275. 543-547 (1997)
[Publications] Takahashi S., Onodera K., Motohashi H., Suwabe N., Hayashi N., Yanai N., Nabeshima Y., Yamamoto M.: "Arrest in primitive erythroid cell development caused by promoter-specific distruption of the GATA-1 gene." gene.J.Biol.Chem.272. 12611-12615 (1997)
[Publications] Sakamoto K., Yan L., Imai H., Takagi M., Nabeshima Y., Takeda S., Katsube K.: "Identifiation of a chick homologue of Fringe,C-Fringe1.Involvement in the neurogenesis and the somitogenesis." Biochem.Biophys.Res.Comm.234. 754-759 (1997)
[Publications] Itoh K., Chiba T., Takahashi S., Ishii T., Igarashi K., Katoh Y., Oyake T., Hayashi N., Satoh K., Yamamoto M., Nabeshima Y.: "An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying genes through antioxidant response elements." Biochem.Biophys.Res.Comm.236. 313-322 (1997)
[Publications] Miyagoe Y., Terashima T., Nonaka I., Hanaoka K., Hayasaka M., Nabeshima Y., ArahataK., Nabeshima Y.,and Takeda S.: "Liminin α2 chain-null mutant mice by targeted disuruption of the Lama2 gene: A new model of merosin (laminin 2)-deficient congenital muscular dystrophy." FEBS Letters. 415. 33-39 (1997)
[Publications] Kuro-o M., Matusumura Y., Aizawa H., Kawaguchi H., Suga T., Utsugi T., Ohyama Y., Kaname T., Kume E., Iwasaki H., Iida A., Shiraki-Iida T., Nishikawa S., Nagai R.and Nabeshima Y.: "Mutation of the mouse Klotho gene leads to a syndrome resembling ageing." Nature. 390. 45-51 (1997)
[Publications] Kameya S., Araki E., Katsuki M., Mizota A., Adachi E., Nakahara K., Nonaka I.,Sakuragi S., Takeda S.,and Nabeshima Y.: "Dp260 dusrupted mice revealed prolonged implicit time of the b-wave in ERG and loss of accumulation of β-dystroglycan in the outer plexiform layer of retina." Human Molocular Genetics. 6. 2195-2203 (1997)
[Publications] Ikeshima-Kataoka H., Skeath J.B., Nabeshima Y., Doe C.Q.,and Mtsuzaki F.: "Miranda directs Prospero to a daughter cell during Drosophila asymmeteric divisions." Nature. 390. 625-629 (1997)
[Publications] Yoshida N., Yoshida S., Fujisawa-Sehara A.,and Nabeshima Y.: "Variation of MyoD and myf-5,which presedes myogenin expression and irreversible commitment,specifies the differentiating and“Reserve Cells " upon myogenesis." J.Cell Sci.(in press).
[Publications] MatsumuraY., Aizawa H., Shiraki-Iida T., Nagai R., Kuro-o M., Nabeshima Y.: "Identification of the human klotho gene and its two transcripts encoding membrane and secreted Koltho protein." B.B.R.C. (in press).
[Publications] Shiraki-Iida T., Aizawa H., Matsumura Y., Sekine S., Iida A., Anazawa H., Hagai R., Kuro-o M., Nabeshima Y.: "Structure of the mouse Klotho gene and its two transcripts encoding membrane and secreted protein." FEBS Letters. (in press).
[Publications] Yamashita T., Nifuzi A., Furuya K.,Nabeshima Y., Noda M.: "Elongation of the epiphyseal trabecular bon associated with precocious aging in transgenic mice carrying an insertional mutation in Klotho gene locus." J.Bone Mineral Res.(in press).
[Publications] Okuda A., Fukushima A., Nishimoto M., Nabeshima Y., Kuro-o M., Nabeshima Y., Boon K.,Keaveney M., Stunnenberg H.G., Muramatsu M.: "UTF1,a transcriptional coacruvator representing part of the E1A-like activity in plutripotent embryonic stem cells." EMBOJ.(in press).

1997 Fiscal Year Annual Research Report

筋細胞系をモデルとした細胞増殖と分化の制御機構の研究

Principal Investigator

鍋島 陽一 国立精神・神経センター, 神経研究所・遺伝子工学, 部長 (60108024)

Research Products

[Publications] Yoshida S., FujisawaーSehara., Arai K. and Nabeshima Y.: "Lysophosphatidic acid and bFGF control different modes in proliferating myoblasts." J.Cell.Biol.132. 181-193 (1996)

[Publications] Hoshino M., Suzuki E., Nabeshima Y. and Hama C.: "hikaru genki protein secreted into synaptic cleft from an early stage of synapse formation in Drosopiola." Development. 122. 589-597 (1996)

[Publications] Ohbayashi T., Schmidt E.E., Makino Y., Kishimoto T., Nabeshima Y., Muramatsu M. and Tamura T.: "Promotor structure of the mouse TATA-binding protein (TBP)gene." Biochem.Biophys.Res.Comm.225. 257-280 (1996)

[Publications] Sone M., Hoshino M., Suzuki E., Saigo K., Nabeshima Y. and Hama C.: "Still life,a protein in synaptic tereminals of Drosophila homologous to GDP-GTP exchanger." Science. 275. 543-547 (1997)

[Publications] Takahashi S., Onodera K., Motohashi H., Suwabe N., Hayashi N., Yanai N., Nabeshima Y., Yamamoto M.: "Arrest in primitive erythroid cell development caused by promoter-specific distruption of the GATA-1 gene." gene.J.Biol.Chem.272. 12611-12615 (1997)

[Publications] Sakamoto K., Yan L., Imai H., Takagi M., Nabeshima Y., Takeda S., Katsube K.: "Identifiation of a chick homologue of Fringe,C-Fringe1.Involvement in the neurogenesis and the somitogenesis." Biochem.Biophys.Res.Comm.234. 754-759 (1997)

[Publications] Kuro-o M., Matusumura Y., Aizawa H., Kawaguchi H., Suga T., Utsugi T., Ohyama Y., Kaname T., Kume E., Iwasaki H., Iida A., Shiraki-Iida T., Nishikawa S., Nagai R.and Nabeshima Y.: "Mutation of the mouse Klotho gene leads to a syndrome resembling ageing." Nature. 390. 45-51 (1997)

[Publications] Ikeshima-Kataoka H., Skeath J.B., Nabeshima Y., Doe C.Q.,and Mtsuzaki F.: "Miranda directs Prospero to a daughter cell during Drosophila asymmeteric divisions." Nature. 390. 625-629 (1997)

[Publications] Yoshida N., Yoshida S., Fujisawa-Sehara A.,and Nabeshima Y.: "Variation of MyoD and myf-5,which presedes myogenin expression and irreversible commitment,specifies the differentiating and“Reserve Cells " upon myogenesis." J.Cell Sci.(in press).

[Publications] MatsumuraY., Aizawa H., Shiraki-Iida T., Nagai R., Kuro-o M., Nabeshima Y.: "Identification of the human klotho gene and its two transcripts encoding membrane and secreted Koltho protein." B.B.R.C. (in press).

[Publications] Shiraki-Iida T., Aizawa H., Matsumura Y., Sekine S., Iida A., Anazawa H., Hagai R., Kuro-o M., Nabeshima Y.: "Structure of the mouse Klotho gene and its two transcripts encoding membrane and secreted protein." FEBS Letters. (in press).

[Publications] Yamashita T., Nifuzi A., Furuya K.,Nabeshima Y., Noda M.: "Elongation of the epiphyseal trabecular bon associated with precocious aging in transgenic mice carrying an insertional mutation in Klotho gene locus." J.Bone Mineral Res.(in press).

[Publications] Okuda A., Fukushima A., Nishimoto M., Nabeshima Y., Kuro-o M., Nabeshima Y., Boon K.,Keaveney M., Stunnenberg H.G., Muramatsu M.: "UTF1,a transcriptional coacruvator representing part of the E1A-like activity in plutripotent embryonic stem cells." EMBOJ.(in press).

鍋島陽一国立精神・神経センター, 神経研究所・遺伝子工学, 部長 (60108024)