Alvarado Jorge A, Betanzos Abigail, Franse-Carman Linda, Chen Janet, González-Mariscal Lorenza
Department of Ophthalmology, University of California, San Francisco, California 94143-0730, USA.
Am J Physiol Cell Physiol. 2004 Mar;286(3):C621-34. doi: 10.1152/ajpcell.00108.2003. Epub 2003 Nov 12.
The purpose of this study was to compare human endothelial cells from Schlemm's canal (SCEs) and the trabecular meshwork (TMEs) in terms of ZO-1 isoform expression, hydraulic conductivity (HC) properties, and "giant" vacuole (GV) formation. The principal study methods were Western blot, RT-PCR, immunofluorescence, and perfusion chambers. Blot signals for alpha+ - and alpha- -isoforms were similar in SCEs but less intense for the alpha+ -relative to the alpha- -signal in TMEs. With the anti-alpha+ antibody used at 1/50 dilution, binding occurred at cell borders of both cell types, but only to SCEs when used at a >/=1/200 dilution in vitro and in vivo. SCEs were more resistive than TMEs (HC = 0.66 vs. 1.32 microl.min-1.mmHg-1.cm-2; P < 0.001) when perfused from apex to base. When perfused in the other direction, SCEs were again more resistive (5.23 vs. 9.04 microl.min-1.mmHg-1.cm-2; P < 0.01). GV formation occurred only in SCEs as a function of flow direction, perfusion pressure, and time. We conclude that SCEs and TMEs have distinctive phenotypic properties involving their content of ZO-1 isoforms, barrier function, and GV formation.
本研究的目的是比较施莱姆管内皮细胞(SCEs)和小梁网内皮细胞(TMEs)在紧密连接蛋白1(ZO-1)亚型表达、水力传导率(HC)特性和“巨型”液泡(GV)形成方面的差异。主要研究方法包括蛋白质免疫印迹法、逆转录-聚合酶链反应(RT-PCR)、免疫荧光法和灌注室法。SCEs中α + -和α - -亚型的印迹信号相似,但相对于TMEs中的α - -信号,TMEs中α + -的信号强度较弱。使用1/50稀释度的抗α + 抗体时,两种细胞类型的细胞边界均出现结合,但在体外和体内使用≥1/200稀释度时,仅SCEs出现结合。从顶端向基部灌注时,SCEs比TMEs的阻力更大(HC = 0.66对1.32微升·分钟-1·毫米汞柱-1·厘米-2;P < 0.001)。当向相反方向灌注时,SCEs的阻力仍然更大(5.23对9.04微升·分钟-1·毫米汞柱-1·厘米-2;P < 0.01)。GV的形成仅发生在SCEs中,是流动方向、灌注压力和时间的函数。我们得出结论,SCEs和TMEs具有独特的表型特性,包括它们的ZO-1亚型含量、屏障功能和GV形成。