Inhibition of RhoA signaling with increased Bves in trabecular meshwork cells.
Inhibition of RhoA signaling with increased Bves in trabecular meshwork cells.
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DOI：
10.1167/iovs.09-3539
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发表时间：
2010-01
影响因子：
4.4
通讯作者：
Chang MS
中科院分区：
文献类型：
--
作者：
Russ PK;Kupperman AI;Presley SH;Haselton FR;Chang MS
血管心外膜物质（Bves）是一种新型的粘附分子，调节紧密连接（TJ）的形成。TJ还调节RhoA信号传导，这与流出调节有关。鉴于Bves已在多种眼部组织中报道，作者假设Bves在小梁网（TM）细胞中的RhoA信号传导调节中发挥作用。评价过表达Bves的人TM细胞系NTM-5和NTM-5（NTM-w）的TJ形成，并比较闭合蛋白、扣带蛋白和ZO-1蛋白的水平。使用荧光素钠扩散和跨细胞电阻（TER）进行TJ功能测定。使用FRET探针测量活化的RhoA的水平，并通过Western印迹分析评估RhoA的下游靶标肌球蛋白轻链（MLC-p）的磷酸化。Bves的过表达导致NTM-5细胞中TJ形成增加。增加的闭合蛋白、扣带蛋白和ZO-1蛋白证实了TJ形成增加。在功能上，与NTM-5细胞相比，NTM-w细胞显示出降低的渗透性和增加的TER，与增加的TJ形成一致。与NTM-5细胞相比，NTM-w细胞还表现出活性RhoA水平降低和MLC-p水平降低。这些发现支持TJ在RhoA信号传导中的作用。TM细胞中Bves的增加导致TJ形成增加，RhoA活化降低，MLC-p降低。这是首次报道TM细胞中RhoA上游的调节途径。在TM组织中，RhoA参与流出调节;因此，Bves可能是水流出的关键调节分子。
Blood vessel epicardial substance (Bves) is a novel adhesion molecule that regulates tight junction (TJ) formation. TJs also modulate RhoA signaling, which has been implicated in outflow regulation. Given that Bves has been reported in multiple ocular tissues, the authors hypothesize that Bves plays a role in the regulation of RhoA signaling in trabecular mesh-work (TM) cells. Human TM cell lines NTM-5 and NTM-5 transfected to overexpress Bves (NTM-w) were evaluated for TJ formation, and levels of occludin, cingulin, and ZO-1 protein were compared. Assays of TJ function were carried out using diffusion of sodium fluorescein and transcellular electrical resistance (TER). Levels of activated RhoA were measured using FRET probes, and phosphorylation of myosin light chain (MLC-p), a downstream target of RhoA, was assessed by Western blot analysis. Overexpression of Bves led to increased TJ formation in NTM-5 cells. Increased TJ formation was confirmed by increased occludin, cingulin, and ZO-1 protein. Functionally, NTM-w cells showed decreased permeability and increased TER compared with NTM-5 cells, consistent with increased TJ formation. NTM-w cells also exhibited decreased levels of active RhoA and lower levels of MLC-p than did NTM-5 cells. These findings support a TJ role in RhoA signaling. Increased Bves in TM cells leads to increased TJ formation with decreased RhoA activation and decreased MLC-p. This is the first report of a regulatory pathway upstream of RhoA in TM cells. In TM tissue, RhoA has been implicated in outflow regulation; thus, Bves may be a key regulatory molecule in aqueous outflow.
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DOI：
10.1006/bbrc.2001.5751
发表时间：
2001-10-26
影响因子：
3.1
作者：
Fukiage, C;Mizutani, K;Shearer, TR
通讯作者：
Shearer, TR
影响因子：
3.7
作者：
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影响因子：
11.8
作者：
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Matter, K
影响因子：
2.7
作者：
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通讯作者：
Gilles, Christine
DOI：
10.1073/pnas.0506095103
发表时间：
2006-01-17
影响因子：
11.1
作者：
Ripley, AN;Osler, ME;Bader, D
通讯作者：
Bader, D