N-cadherin-mediated cell adhesion determines the plasticity for cell alignment in response to mechanical stretch in cultured cardiomyocytes
N-cadherin-mediated cell adhesion determines the plasticity for cell alignment in response to mechanical stretch in cultured cardiomyocytes
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DOI：
10.1016/j.bbrc.2004.11.019
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发表时间：
2005-01-07
影响因子：
3.1
通讯作者：
Azuma, J
中科院分区：
文献类型：
--
作者：
Matsuda, T;Takahashi, K;Azuma, J
机械拉伸被认为是心脏的生长刺激。在生理学上，机械牵张有助于心肌细胞的定向，但其分子机制仍有待阐明。本研究旨在阐明N-钙粘蛋白在机械牵张下细胞排列可塑性中的功能意义。新生大鼠心肌细胞，培养在硅胶盘，进行人工单轴周期性拉伸。在接种后的某些时间（3-75小时）开始机械拉伸并持续24小时。培养后3 h的牵张刺激促进细胞取向与张力方向平行。相反，心肌细胞在培养后24-75小时暴露于拉伸时未能对齐。为了阐明N-钙粘蛋白在牵张反应中的重要性，分析了N-钙粘蛋白的表达和分布。接种后即刻，N-钙粘蛋白呈分散分布。在培养过程中，N-钙粘蛋白定位于细胞-细胞接触伴随着其蛋白质的上调。接下来，为了研究细胞-细胞粘附的影响，通过胰蛋白酶处理将培养72小时的心肌细胞重新铺板，并在重新铺板后3小时暴露于拉伸。将胰蛋白酶消化后重新铺片的心肌细胞定向为平行于机械拉伸的张力方向。最后，腺病毒转染的显性负N-钙粘蛋白恢复的能力，表现出细胞的方向响应拉伸。结果提示，N-钙粘蛋白参与了机械牵张诱导的心肌细胞定向反应。(C)2004年爱思唯尔公司All rights reserved.
Mechanical stretch has been implicated as the growth stimuli in the heart. Physiologically, mechanical stretch is reported to contribute to the orientation of cardiomyocytes, though the molecular mechanism remains to be elucidated. This study was designed to make clear functional significances of N-cadherin in plasticity of cell alignment in response to mechanical stretch. Neonatal rat cardiomyocytes, cultured on silicone dishes, were subjected to artificial uniaxial cyclic stretch. Mechanical stretch was started at certain times (3-75 h) after seeding and continued for 24 h. Stretch stimulation in 3 h after cultivation promoted cell orientation running parallel to tension direction. In contrast, cardiac myocytes fail to align when exposed to stretch 24-75 h after cultivation. To address the importance of N-cadherin in the responsiveness to stretch, the expression and distribution of N-cadherin were analyzed. Immediately after seeding, N-cadherin showed dispersed distributions. During cultivation, N-cadherin localized to cell-cell contacts accompanied by the upregulation of its protein. Next, to investigate influence of cell-cell adhesion, cardiomyocytes cultured for 72 h were replated by trypsin treatment and exposed to stretch 3 h after replating. The cardiomyocytes replated by trypsinization were oriented in parallel to tension direction by mechanical stretch. Finally, adenoviral transfection of dominant-negative N-cadherin recovered the ability to exhibit cell orientation in response to stretch. Our results suggested that N-cadherin was involved in the oriented responses of cardiomyocytes induced by mechanical stretch. (C) 2004 Elsevier Inc. All rights reserved.