1998 Fiscal Year Annual Research Report

成熟心筋細胞収縮機能調整に関する受容体・細部内情報伝達分子機構の解明-組換えアデノウイルスを用いて遺伝子導入した単一成熟心筋細胞を用いた生理機能解析-

Research Project

Project/Area Number	10877112
Research Institution	Kyushu University
Principal Investigator	上野光九州大学, 医学部, 講師 (50260378)
Co-Investigator(Kenkyū-buntansha)	佐藤真司九州大学, 医学部, 助手 (60274445)
Keywords	単一心筋細胞 / スキンド細胞 / 遺伝子導入 / 張力測定 / GTP結合蛋白質 / アドレノセプター / アデノウイルス
Research Abstract	1.ラット心室筋細胞を単離し、β-escinにて細胞膜に微小な穴をあけたスキンド細胞を準備した。この標本では細胞内のカルシウム濃度を自由に設定することができる。さらにこの単一心筋スキンド細胞の収縮を経時的に測定できることを確認した。 2.上記のスキンド細胞をベータアゴニストで刺激すると、収縮蛋白質のカルシウム感受性が低下した。 3.一方、アルファアゴニストで刺激すると、収縮蛋白質のカルシウム感受性は亢進し、筋小胞体からのカルシウム放出が惹起された。 4.上記の反応は1mMのGDPβbSで抑制された。従ってGTP結合蛋白質の活性化を介していることが示唆される。 5.以上の結果は、受容体機能さらに受容体下流の信号伝達系の少なくとも一部(GTP結合蛋白質など)を保持したまま微小孔から細胞内のカルシウム環境を自由に変化させながら収縮力の発生を測定できる実験系が確立されたことを意味している。 6.心筋細胞でのPharmaco-mechanicalcouplingにおけるGTP結合蛋白質(Ras,Rac,Rho,Cdc42)の役割を解明するためにこれらの分子機能を特異的に阻害するそれぞれのDominant-negative変異体を作成し、さらにアデノウイルスベクターへの組み込みに成功した。今年度は作製したアデノウイルスを使ってこれらの分子を心筋細胞に遺伝子発現させ、GTP結合蛋白質の収縮力発生における役割を解明する予定である。

Research Products
(11 results)

All Other

All Publications (11 results)

[Publications] Sato S,et al.: "Adrenoceptor-mediated regulation of myofibrillar Ca sensitivity through the GTP-binding protein-related mechanisms: tension recording in b-escub-skinned single rat cardiac cells with preserved receptor functions." Eur J Physiol.in press.
[Publications] Matsumoto T,et al.: "Pletelet-derived growth factor activates p38 mitogen activated protein kinase through ras-dependent pathway shiech is important for actin reoganization and cell miaration." Journal of Biological Chemistry. in press.
[Publications] Qi Z,et al.: "Blockade of TGF-β signaling prevents liver fibrosis and dysfunction." Proc.Natl.Acad.Sci.USA.in press.
[Publications] Yamamoto H,et al.: "Roles of H-ras in signaling by TGF-β:H-ras is essential in activation of MAP kinase,partially involved in transcriptional activation by TGF-β but not in signaling of anti-proliferative" Eur.J.Biochem.in press.
[Publications] Ohashi M,et al.: "Adenovirus-mediated p53 tumor suppressor gene therapy of human gastric cancer cells in vitro and in vivo." Gut. in press.
[Publications] Yukawa H,et al.: "In vitro growth suppression of vascular smooth muscle cells using adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor." Atherosclerosis. in press.
[Publications] Sakamoto T,et al.: "Transfection of LacZ gene to retina by an adenoviral vector causes retinal dysfunction with no apparent morphological changes." Hum.Gene Ther.9. 789-799 (1998)
[Publications] Takayama K,et al.: "The levels of integrin α V and β5 may well predict the susceptibility to adenovirus-mediated gene transfer in human lung cancer cells." Gene Ther.5. 361-368 (1998)
[Publications] Sakamoto T,et al.: "A vitrctomy improves the transfection efficiency of adenoviral vector-mediated gene transfer to Muller cells." Gene Ther.5. 1088-1097 (1998)
[Publications] Li Q,et al.: "Stretch-induced collagen synthesis in cultured smooth muscle cells from rabbit aortic media and possible involvment of angiotensin and TGF-β." J.Vasc.Res.35. 93-103 (1998)
[Publications] Maru Y,et al.: "Virally activated Res cooperates wit integrin to induce tubulogenesis in sinusoidal endothelial cell lines." J.Cell.Physiol.176. 223-234 (1998)

1998 Fiscal Year Annual Research Report

成熟心筋細胞収縮機能調整に関する受容体・細部内情報伝達分子機構の解明-組換えアデノウイルスを用いて遺伝子導入した単一成熟心筋細胞を用いた生理機能解析-

Principal Investigator

上野 光 九州大学, 医学部, 講師 (50260378)

Research Products

[Publications] Sato S,et al.: "Adrenoceptor-mediated regulation of myofibrillar Ca sensitivity through the GTP-binding protein-related mechanisms: tension recording in b-escub-skinned single rat cardiac cells with preserved receptor functions." Eur J Physiol.in press.

[Publications] Matsumoto T,et al.: "Pletelet-derived growth factor activates p38 mitogen activated protein kinase through ras-dependent pathway shiech is important for actin reoganization and cell miaration." Journal of Biological Chemistry. in press.

[Publications] Qi Z,et al.: "Blockade of TGF-β signaling prevents liver fibrosis and dysfunction." Proc.Natl.Acad.Sci.USA.in press.

[Publications] Yamamoto H,et al.: "Roles of H-ras in signaling by TGF-β:H-ras is essential in activation of MAP kinase,partially involved in transcriptional activation by TGF-β but not in signaling of anti-proliferative" Eur.J.Biochem.in press.

[Publications] Ohashi M,et al.: "Adenovirus-mediated p53 tumor suppressor gene therapy of human gastric cancer cells in vitro and in vivo." Gut. in press.

[Publications] Yukawa H,et al.: "In vitro growth suppression of vascular smooth muscle cells using adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor." Atherosclerosis. in press.

[Publications] Sakamoto T,et al.: "Transfection of LacZ gene to retina by an adenoviral vector causes retinal dysfunction with no apparent morphological changes." Hum.Gene Ther.9. 789-799 (1998)

[Publications] Takayama K,et al.: "The levels of integrin α V and β5 may well predict the susceptibility to adenovirus-mediated gene transfer in human lung cancer cells." Gene Ther.5. 361-368 (1998)

[Publications] Sakamoto T,et al.: "A vitrctomy improves the transfection efficiency of adenoviral vector-mediated gene transfer to Muller cells." Gene Ther.5. 1088-1097 (1998)

[Publications] Li Q,et al.: "Stretch-induced collagen synthesis in cultured smooth muscle cells from rabbit aortic media and possible involvment of angiotensin and TGF-β." J.Vasc.Res.35. 93-103 (1998)

[Publications] Maru Y,et al.: "Virally activated Res cooperates wit integrin to induce tubulogenesis in sinusoidal endothelial cell lines." J.Cell.Physiol.176. 223-234 (1998)

上野光九州大学, 医学部, 講師 (50260378)