...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Investigative ophthalmology & visual science >MicroRNA-204/211 Regulates Human Retinal Pigment Epithelial Physiology
【24h】

MicroRNA-204/211 Regulates Human Retinal Pigment Epithelial Physiology

机译：MicroRNA-204 / 211调节人类视网膜色素上皮生理

获取原文
   

获取外文期刊封面封底 >>

       
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Purpose: : To determine a set of microRNAs that are enriched in human RPE compared to retina and choroid. Using a human fetal retinal pigment epithelial (hfRPE) culture model, we studied the regulation of miR-204/211 on RPE physiology. Methods: : Primary hfRPE cultures were obtained as described previously. miRNA in human RPE, retina and choroid were profiled using the TaqMan?? MicroRNA Assays Human Panel Early Access Kit. Q-RT-PCR was carried out with TaqMan?? microRNA assay. miRNA Northern blots were done with the LNATM probes and in situ hybridization was performed with digoxigenin (Dig)-labeled probes. Direct miRNA targets were determined by a dual luciferase reporter assay using pEZX-MT01- target constructs. RPE physiology on intact monolayers of hfRPE was studied as previously described. Results: : We determined eight miRNAs that are enriched (10 fold) in RPE (miRs-184,187, 200a/200b, 204/211, 221/222) compared to neuroretina or choroid (p0.05). Five of these miRNAs are enriched in RPE compared to 20 tissues throughout the body and are 10,000 fold more highly expressed (p0.005) and miR-204/211 are the most highly expressed. We determined that TGF?2R2 and Snail2 are direct targets of miR-204. Addition of anti-miR-204 significantly decreased: (1) transepithelial resistance (TER); (2) cell membrane potentials; (3) claudin expression (RNA and protein); (4) Kir 7.1 (K channels) (protein). Conclusions: : miR-204 regulation of RPE membrane physiology is mediated through a TGF?2 /SNAI2 signaling pathway that controls the expression of claudins in the tight junctions and Kir 7.1 in the plasma membranes. The present experiments provide the basis for understanding how miR-204/211 regulates RPE tight junction integrity and maintain the blood/retina barrier in a quiescent state, which is a hallmark of epithelia.
机译：目的：：确定一组与视网膜和脉络膜相比富含人RPE的microRNA。使用人类胎儿视网膜色素上皮（hfRPE）培养模型，我们研究了miR-204 / 211对RPE生理的调节。方法：：如前所述获得原代hfRPE培养物。使用TaqMan ??分析了人类RPE，视网膜和脉络膜中的miRNA。 MicroRNA分析人类面板早期获取试剂盒。用TaqMan ??进行Q-RT-PCR。 microRNA分析。使用LNATM探针进行miRNA Northern印迹，并用洋地黄毒苷（Dig）标记的探针进行原位杂交。直接的miRNA靶标是使用pEZX-MT01-靶标构建体通过双重萤光素酶报告基因测定的。如前所述，研究了hfRPE完整单层上的RPE生理学。结果：与神经视网膜或脉络膜相比，我们确定了八个在RPE中富集（> 10倍）的miRNA（miRs-184,187、200a / 200b，204 / 211、221 / 222）（p <0.05）。与整个身体的20个组织相比，这些miRNA中有五个在RPE中富集，并且高表达> 10,000倍（p <0.005），而miR-204 / 211则是最高表达的。我们确定TGFβ2R2和Snail2是miR-204的直接靶标。抗miR-204的添加明显减少：（1）跨上皮抵抗（TER）; （2）细胞膜电位; （3）claudin表达（RNA和蛋白质）; （4）Kir 7.1（K通道）（蛋白质）。结论：miR-204对RPE膜生理的调节是通过TGFβ2/ SNAI2信号通路介导的，该信号通路控制紧密连接中claudins的表达和质膜Kir 7.1的表达。本实验为理解miR-204 / 211如何调节RPE紧密连接的完整性并在静止状态维持血液/视网膜屏障提供了基础，这是上皮细胞的标志。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号