Degradation of filamin induces contraction of vascular smooth muscle cells in Type-I collagen matrix honeycombs.
Degradation of filamin induces contraction of vascular smooth muscle cells in Type-I collagen matrix honeycombs.
复制标题
细丝蛋白的降解诱导 I 型胶原基质蜂窝中血管平滑肌细胞的收缩。
DOI：
10.1159/000330076
复制
发表时间：
2011
影响因子：
--
通讯作者：
Uchida M Ishii I Hirata K Yamamoto F Tashiro K Suzuki T Nakayama Y Ariyoshi N and Kitada M.
中科院分区：
文献类型：
--
作者：
Tomizawa;A.;Hadjidekov;G.;Ishii;I.. Bakalova;R.;Zhelev;Z.;Aoki;I.;Saga;T.;Kitada;M.;Uchida M Ishii I Hirata K Yamamoto F Tashiro K Suzuki T Nakayama Y Ariyoshi N and Kitada M.
背景资料：去分化的兔血管平滑肌细胞（SMC）在称为“蜂窝”的I型胶原三维基质中培养时表现出与分化的SMC相似的特征，但其机制尚不清楚。细丝蛋白是一种肌动蛋白结合蛋白，在平滑肌细胞中连接肌动蛋白丝。方法：采用免疫印迹法和免疫荧光染色法检测细丝蛋白等相关蛋白的表达。测量蜂窝尺寸以确认SMC的收缩。结果如下：全长细丝蛋白表达在亚汇合SMC培养板上，然而，细丝蛋白的降解，这可能是由钙蛋白酶调节，观察到汇合SMC培养板上和蜂窝。虽然细丝蛋白与β-肌动蛋白共定位在平板上生长的亚融合SMC中，但在蜂窝中培养的SMC的细胞质中检测到细丝蛋白，并且降解的细丝蛋白主要在这些细胞的细胞质部分中检测到。此外，与在平板上培养的细胞相比，在蜂窝中培养的SMC的细胞骨架部分中β-肌动蛋白表达较低，并且用于培养SMC的蜂窝的大小显著减小。结论：这些数据表明，在蜂窝中培养的SMC中细丝蛋白的降解诱导β-非肌肉肌动蛋白细丝的结构弱化，从而允许蜂窝中的SMC实现收缩性。
Background: Dedifferentiated rabbit vascular smooth muscle cells (SMCs) exhibit similar features to differentiated SMCs when cultured in three-dimensional matrices of type-I collagen called “honeycombs,” but the mechanism is unknown. The role of filamin, an actin-binding protein that links actin filaments in SMCs, was investigated. Methods: Filamin and other related proteins were detected by western blot analysis and immunofluorescence staining. Honeycomb size was measured to confirm the contraction of SMCs. Results: Full-length filamin was expressed in subconfluent SMCs cultured on plates; however, degradation of filamin, which might be regulated by calpain, was observed in confluent SMCs cultured on plates and in honeycombs. While filamin was co-localized with β-actin in subconfluent SMCs grown on plates, filamin was detected in the cytoplasm in SMCs cultured in honeycombs, and degraded filamin was mainly detected in the cytoplasmic fraction of these cells. In addition, β-actin expression was low in the cytoskeletal fraction of SMCs cultured in honeycombs compared with cells cultured on plates, and the size of the honeycombs used for culturing SMCs was significantly reduced. Conclusion: These data suggest that degradation of filamin in SMCs cultured in honeycombs induces structural weakness of β-non-muscle actin filaments, thereby permitting SMCs in honeycombs to achieve contractility.