Chloride intracellular channel 4 is critical for the epithelial morphogenesis of RPE cells and retinal attachment.
Chloride intracellular channel 4 is critical for the epithelial morphogenesis of RPE cells and retinal attachment.
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DOI：
10.1091/mbc.e09-10-0907
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发表时间：
2010-09-01
影响因子：
3.3
通讯作者：
Sung CH
中科院分区：
文献类型：
--
作者：
Chuang JZ;Chou SY;Sung CH
使用基于质粒的转染方法原位细胞自主沉默 RPE 中的氯离子胞内通道 4 (CLIC4)。这些结果表明 CLIC4 对于上皮形态发生和视网膜附着至关重要。视网膜脱离治疗的新候选靶标也已确定。视网膜脱离是一种威胁视力的疾病。 RPE 和光感受器之间粘附的分子机制尚不清楚，因为这两种细胞类型之间的密切相互作用不可能在体外进行建模和研究。在本文中，我们发现氯细胞内通道 4 (CLIC4) 在顶端 RPE 微绒毛处富集，这些微绒毛与光感受器外节相互交叉。我们使用一种新的基于质粒的转染方法来原位细胞自主抑制 RPE 中的 CLIC4。 CLIC4沉默的RPE细胞表现出顶端微绒毛和基底内皱的显着损失、视网膜粘附力的降低和上皮间质转化。异位表达的ezrin未能挽救CLIC4沉默所造成的形态变化。与 CLIC4 抑制的 RPE 细胞相邻的神经视网膜表现出严重的发育不良。最后，CLIC4 抑制的 RPE 细胞的顶端表面意外地出现了高水平的水通道蛋白 1，同时单羧酸转运蛋白 MCT3 的基底表面表达也随之丧失。我们的结果表明 CLIC4 在 RPE-光感受器粘附中发挥重要作用，可能是通过调节细胞表面通道/转运蛋白的活性。我们认为这些变化可能归因于我们新型视网膜脱离动物模型中的视网膜下液积聚。
A plasmid-based transfection method was used to cell-autonomously silence chloride intracellular channel 4 (CLIC4) in RPE in situ. These results show CLIC4 is critical for epithelial morphogenesis and retinal attachment. Novel candidate targets for retinal detachment therapy have also been identified. Retinal detachment is a sight-threatening condition. The molecular mechanism underlying the adhesion between the RPE and photoreceptors is poorly understood because the intimate interactions between these two cell types are impossible to model and study in vitro. In this article, we show that chloride intracellular channel 4 (CLIC4) is enriched at apical RPE microvilli, which are interdigitated with the photoreceptor outer segment. We used a novel plasmid-based transfection method to cell-autonomously suppress CLIC4 in RPE in situ. CLIC4 silenced RPE cells exhibited a significant loss of apical microvilli and basal infoldings, reduced retinal adhesion, and epithelial-mesenchymal transition. Ectopically expressing ezrin failed to rescue the morphological changes exerted by CLIC4 silencing. Neural retinas adjacent to the CLIC4-suppressed RPE cells display severe dysplasia. Finally, a high level of aquaporin 1 unexpectedly appeared at the apical surfaces of CLIC4-suppressed RPE cells, together with a concomitant loss of basal surface expression of monocarboxylate transporter MCT3. Our results suggested that CLIC4 plays an important role in RPE-photoreceptor adhesion, perhaps by modulating the activity of cell surface channels/transporters. We propose that these changes may be attributable to subretinal fluid accumulation in our novel retinal detachment animal model.