Regulation of cross-linked actin network (CLAN) formation in human trabecular meshwork (HTM) cells by convergence of distinct beta1 and beta3 integrin pathways.
Regulation of cross-linked actin network (CLAN) formation in human trabecular meshwork (HTM) cells by convergence of distinct beta1 and beta3 integrin pathways.
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DOI：
10.1167/iovs.08-3215
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发表时间：
2009-12
影响因子：
4.4
通讯作者：
Peters DM
中科院分区：
文献类型：
--
作者：
Filla MS;Schwinn MK;Sheibani N;Kaufman PL;Peters DM
确定β1/β3整联蛋白介导的调节人小梁网（HTM）细胞中交联肌动蛋白网络（CLAN）形成的途径。CLAN在昏迷和类固醇处理的TM细胞中形成，这可能有助于减少通过TM的流出设施。通过FACS评估CD 47（αvβ3整合素共受体/血小板反应蛋白-1受体）以及整合素αvβ3和β1的表达。在不存在或存在β3整联蛋白活化mAb AP-5的情况下，通过将细胞接种在纤连蛋白（β1整联蛋白配体）上诱导CLAN，并通过鬼笔环肽标记进行鉴定。通过将细胞与抑制剂PP 2和EPA（Src激酶）、LY 294002（PI-3 K）或NSC 23766（Rac 1）一起孵育来确定Src激酶、PI-3激酶（PI-3 K）、Rac 1和CD 47的作用。Tiam 1和Trio siRNA以及显性阴性Tiam 1用于确定涉及哪种Rac 1特异性鸟嘌呤核苷酸交换因子。使用血小板反应蛋白-1衍生的激动剂肽4 N1 K和CD 47功能阻断抗体B6H12.2确定CD 47的作用。HTM细胞表达CD 47和整合素αvβ3和β1。与β1整联蛋白诱导水平相比，β3整联蛋白或CD 47活化显著增加了CLAN形成，而抗CD 47 mAb B6H12.2抑制了这种增加。PP 2、NSC 23766和Trio siRNA分别使β3诱导的CLAN形成减少72%、45%和67%，而LY 294002和显性阴性Tiam 1没有影响。LY 294002使β1整合素介导的CLAN形成减少42%，PP 2完全阻断它。β1介导的CLAN形成是PI 3-K依赖性的，而β3介导的CLAN形成是CD 47和Rac 1/Trio依赖性的，并且可能受血小板反应蛋白-1调节。两种整合素途径都是Src依赖性的。
Determine the β1/β3 integrin-mediated pathways that regulate cross-linked actin network (CLAN) formation in human trabecular meshwork (HTM) cells. CLANs form in glaucomatous and steroid-treated TM cells which may contribute to reducing outflow facility through the TM. Expression of CD47 (an αvβ3 integrin co-receptor/thrombospondin-1 receptor) and integrins αvβ3 and β1 was assessed by FACS. CLANs were induced by plating cells on fibronectin (a β1 integrin ligand) in the absence or presence of the β3 integrin-activating mAb AP-5 and identified by phalloidin labeling. The role of Src kinases, PI-3 Kinase (PI-3K), Rac1 and CD47 was determined by incubating cells with the inhibitors PP2 and EPA (Src kinases), LY294002 (PI-3K), or NSC23766 (Rac1). Tiam1 and Trio siRNAs and dominant-negative Tiam1 were used to determine which Rac1-specific guanine nucleotide exchange factor was involved. The role of CD47 was determined using the thrombospondin-1-derived agonist peptide 4N1K and the CD47 function blocking antibody B6H12.2. HTM cells expressed CD47 and integrins αvβ3 and β1. β3 integrin or CD47 activation significantly increased CLAN formation over β1 integrin-induced levels while anti-CD47 mAb B6H12.2 inhibited this increase. PP2, NSC23766 and Trio siRNA decreased β3-induced CLAN formation by 72%, 45% and 67% respectively while LY294002 and dominant negative Tiam1 had no effect. LY294002 decreased β1 integrin-mediated CLAN formation by 42% and PP2 completely blocked it. Distinct β1 and αvβ3 integrins signaling pathways converge to enhance CLAN formation. β1-mediated CLAN formation was PI3-K-dependent while β3-mediated CLAN formation was CD47- and Rac1/Trio-dependent and may be regulated by thrombospondin-1. Both integrin pathways were Src-dependent.
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影响因子：
3.4
作者：
Flügel-Koch, C;Ohlmann, A;Tamm, ER
通讯作者：
Tamm, ER
DOI：
10.1083/jcb.135.2.533
发表时间：
1996-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gao AG;Lindberg FP;Dimitry JM;Brown EJ;Frazier WA
通讯作者：
Frazier WA
影响因子：
4.8
作者：
Frazier, WA;Gao, AG;Linder, ME
通讯作者：
Linder, ME
影响因子：
3.4
作者：
Filla, MS;David, G;Peters, DM
通讯作者：
Peters, DM
影响因子：
2.3
作者：
Gelatt, KN;Mackay, EO
通讯作者：
Mackay, EO