Laminin-332 alters connexin profile, dye coupling and intercellular Ca2+ waves in ciliated tracheal epithelial cells.
Laminin-332 alters connexin profile, dye coupling and intercellular Ca2+ waves in ciliated tracheal epithelial cells.
复制标题
层粘连蛋白332在纤毛气管上皮细胞中改变连接蛋白的谱，染料偶联和细胞间Ca2+波。
DOI：
10.1186/1465-9921-7-105
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发表时间：
2006-08-02
影响因子：
5.8
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中科院分区：
文献类型：
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气管上皮细胞锚定在动态基底膜上，该基底膜含有多种细胞外基质蛋白，包括胶原蛋白和层粘连蛋白。在气道上皮的发育、创伤修复和疾病过程中，细胞外基质蛋白的显著变化可能直接影响细胞迁移、分化和由细胞间通讯介导的事件。我们假设细胞基质的改变，特别是基质中的I型胶原和层粘连蛋白α3β3γ2（LM-332）蛋白，直接影响纤毛兔气管上皮细胞（RTEC）的细胞间通讯。通过将带负电荷的荧光染料荧光黄和Alexa 350显微注射到在LM-332/胶原或胶原基质上生长的纤毛RTEC中来监测RTEC的功能偶联。耦合的生理意义的分子进行了评估的机制和程度的传播细胞间Ca 2+波。采用逆转录-聚合酶链反应和免疫细胞化学方法检测细胞间隙连接蛋白（Cx）mRNA和蛋白的表达.当与仅在胶原上生长的RTEC相比时，在LM-332/胶原上生长的RTEC显示出染料转移的显著增加。尽管在LM-332/胶原或单独胶原上生长的RTEC的机械刺激导致细胞间Ca 2+波，但转移机制依赖于基质：在LM-332/胶原上生长的RTEC通过细胞外嘌呤能信号传导传播Ca 2+波，而在胶原上生长的RTEC使用间隙连接。比较在胶原或LM-332/胶原基质上生长的RTEC揭示了Cx 26、Cx43和Cx46蛋白的重组。气道基底膜蛋白（如LM-332）的改变可诱导连接蛋白重组，并导致细胞通讯机制改变，从而影响气道组织功能。
Tracheal epithelial cells are anchored to a dynamic basement membrane that contains a variety of extracellular matrix proteins including collagens and laminins. During development, wound repair and disease of the airway epithelium, significant changes in extracellular matrix proteins may directly affect cell migration, differentiation and events mediated by intercellular communication. We hypothesized that alterations in cell matrix, specifically type I collagen and laminin α3β3γ2 (LM-332) proteins within the matrix, directly affect intercellular communication in ciliated rabbit tracheal epithelial cells (RTEC). Functional coupling of RTEC was monitored by microinjection of the negatively charged fluorescent dyes, Lucifer Yellow and Alexa 350, into ciliated RTEC grown on either a LM-332/collagen or collagen matrix. Coupling of physiologically significant molecules was evaluated by the mechanism and extent of propagated intercellular Ca2+ waves. Expression of connexin (Cx) mRNA and proteins were assayed by reverse transcriptase – polymerase chain reaction and immunocytochemistry, respectively. When compared to RTEC grown on collagen alone, RTEC grown on LM-332/collagen displayed a significant increase in dye transfer. Although mechanical stimulation of RTEC grown on either LM-332/collagen or collagen alone resulted in intercellular Ca2+ waves, the mechanism of transfer was dependent on matrix: RTEC grown on LM-332/collagen propagated Ca2+waves via extracellular purinergic signaling whereas RTEC grown on collagen used gap junctions. Comparison of RTEC grown on collagen or LM-332/collagen matrices revealed a reorganization of Cx26, Cx43 and Cx46 proteins. Alterations in airway basement membrane proteins such as LM-332 can induce connexin reorganizations and result in altered cellular communication mechanisms that could contribute to airway tissue function.