STRETCH-INDUCED CELL MORPHOGENEISI : MOLECULAR MECHANISM OF DIRECTIONSENSITIVITY AND POLARITY FORMATION

拉伸诱导的细胞形态发生：方向敏感性和极性形成的分子机制

• 批准号: 08458206
• 负责人: SOKABE Masahiro
• 金额: $ 5.44万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08458206/
• 关键词: Endothelial cells; Stretch stimuli; Morphological response; Cell polarity; signaling cascade; SA channel; Focal adhesion; Tyrosine phosphorylation; 血管内皮細胞; 一軸伸展刺激; 接着班蛋白質; src; 細胞骨格; 血管内皮細胞、; 機械刺激、; 形態応答、; 機械受容チャネル、; 接着斑、; チロシン燐酸化、

Endothelial cells exhibit spindle like shape aligning their longtude parallel with vessel running. This peculiar shape and alignment is isgnificant to prevent the cells from being pealed off by blood flow. However, cultured endothelial cells do not show such a shape and alignment, It is known that mechanical stresses onto the cells, like shear stress and periodic circumferential stretch, by pulsative blood flow are enough to induce such a morphology in endothelial cells. It has been also suggested that cytoskeletons and adhesion molecules contribute to this morphogenesis. The altimate goal of this project is to elucidate the signaling cascade in the stretchOinduced morphogenesis in cultured endothelial cells and to understand the molecular mechanism underlying the formation of two dimensional polarity in the cell. Using Ca^<2+>imaging, patch clamp, biochemical and molecular biological techniques, we could identify the major signaling cascade in the stretch-induced morphogenesis as follows : <uniaxial periodic stretch* activation of SA channels* intracellular C_amobilization* activation of calcinyrin* activation of tyrosine kinase, src, * tyrosine phosphorylation of adhesion proteins* reorganization of stress fibers and forcal adhesion* morphological change>. However, as the stretch activated Ca^<2+> increase was spatially uniform, this may not be the cause of the cell polarity. On the other hand, tyrosine phosphorylated proteins favored to distribute at the elongating portion of the cell. Hence, it is possible that there is another signaling mechanism flowing from adhesion plaque (integrin), upon which mechanical forces are directly imposed, to intracellular space. This mechanism may along with the former signaling mechanism contribute to the formation of the cell polarity through the regulation of tyrosine phosphorylation of focal adhesion proteins. Next step of our project should be to prove this hypothesis.

内皮细胞表现出纺锤体的形状，将它们的长调与血管的延伸平行。这种特殊的形状和​​比对是可以防止细胞被血流渗出的。然而，培养的内皮细胞没有显示出这种形状和比对，众所周知，通过脉动血流通过剪切应力和周期性圆周拉伸等细胞上的机械应力足以在内皮细胞中诱导这种形态。还建议细胞骨架和粘附分子有助于这种形态发生。该项目的高度目标是阐明培养的内皮细胞中伸展诱导的形态发生中的信号传导级联，并了解细胞中二维极性形成的基础机制。 Using Ca^<2+>imaging, patch clamp, biochemical and molecular biological techniques, we could identify the major signaling cascade in the stretch-induced morphogenesis as follows : <uniaxial periodic stretch* activation of SA channels* intracellular C_amobilization* activation of calcinyrin* activation of tyrosine kinase, src, * tyrosine phosphorylation of adhesion蛋白质*应力纤维和施加粘附的重组*形态变化>。但是，由于拉伸激活的Ca^<2+>在空间上均匀，因此这可能不是细胞极性的原因。另一方面，酪氨酸磷酸化的蛋白质喜欢分布在细胞的细长部分。因此，可能存在另一种信号传导机制，从粘附斑块（整合素）中流动，直接施加机械力到细胞内空间。这种机制可以与以前的信号传导机制一起通过调节局灶性粘附蛋白的酪氨酸磷酸化来促进细胞极性的形成。我们项目的下一步应该是证明这一假设。

项目成果

Suzuki, M., Naruse, K., Asano, Y., Okamoto, T., Nishikimi, N., Sakurai, Y., Nimura, Y., Sokabe, M.: "Up-regulation of integrin beta3 expression by cyclic stretch in human umbilical endothelial cells." Biochem.Biophys.Res.Com.239. 372-376 (1997)

Suzuki, M.、Naruse, K.、Asano, Y.、Okamoto, T.、Nishikimi, N.、Sakurai, Y.、Nimura, Y.、Sokabe, M.：“循环上调整合素 beta3 表达

Naruse, K., Yamada, T., Sokabe, M.: "Involvement of SA channels in orienting response of cultured endothelial cells to cyclic stretch." Am, J.Physiol.(in press).

Naruse, K.、Yamada, T.、Sokabe, M.：“SA 通道参与培养的内皮细胞对循环拉伸的定向反应。”

Sokabe,M., Nunogaki,K. & Naruse,K.: "Strech activated ion channels:Single Channel vs.whole cell responses" Progr Cell Res.6. 139-149 (1997)

索卡部，M.，布垣，K.

Qi Z., Sokabe, M.: "Dynamic properties of individual water molecules in a hydrophobic pore lined with acy1 chains: a molecular dynamics study." Biophys Chem. (in press).

Qi Z.，Sokabe，M.：“内衬 acy1 链的疏水孔中单个水分子的动态特性：分子动力学研究。”

Kobuke,Y.,Tanaka,Y.,Sokabe,M.: "Artificial non-peptide single ion channels." Progr Cell Res.6. 167-188 (1996)

Kobuke,Y.、Tanaka,Y.、Sokabe,M.：“人工非肽单离子通道。”