Bestrophin 2在人无色素睫状上皮中表达，但在视网膜色素上皮中不表达。
Bestrophin 2 is expressed in human non-pigmented ciliary epithelium but not retinal pigment epithelium.
作者信息
Zhang Youwen, Patil Rajkumar V, Marmorstein Alan D
机构信息
Department of Ophthalmology and Vision Science, University of Arizona, Tucson, AZ 85724, USA.
出版信息
Mol Vis. 2010 Feb 10;16:200-6.
PURPOSE
Mice in which bestrophin 2 (Best2) is disrupted exhibit changes in aqueous flow and drainage, resulting in a reduction in intraocular pressure in comparison to wild-type mice. Best2 encodes a putative anion channel localized uniquely to the basolateral plasma membrane of non-pigmented epithelium cells in mice. In this study, we examine the localization of Best2 in the human eye.
METHODS
Rabbit polyclonal antibodies recognizing human Best2 (hBest2) were generated and characterized for use in western blot, immunoprecipitation, and immunofluorescence assays. The expression of hBest2 using these antibodies was examined using human donor eye tissues.
RESULTS
We could not detect hBest2 in human ciliary bodies or other ocular tissues by western blot. However, when enriched by immunoprecipitation, hBest2 was identified in ciliary bodies, but not in the retinal pigment epithelium. Using immunofluorescence, we located hBest2 in the basolateral plasma membrane of non-pigmented epithelial cells.
CONCLUSIONS
We found expression of hBest2 similar to mice only in NPE cells. These data suggest that Best2 may play a functional role in the regulation of aqueous flow and drainage in humans. We conclude that Best2 represents a new potential target for glaucoma therapy.
目的
与野生型小鼠相比，双孔蛋白2（Best2）基因敲除的小鼠房水流动和引流出现变化，导致眼压降低。Best2编码一种推测的阴离子通道，在小鼠中该通道仅特异性定位于非色素上皮细胞的基底外侧质膜。在本研究中，我们检测了Best2在人眼中的定位。
方法
制备并鉴定了识别人类Best2（hBest2）的兔多克隆抗体，用于蛋白质免疫印迹、免疫沉淀和免疫荧光检测。使用这些抗体，利用人类供体眼组织检测hBest2的表达。
结果
通过蛋白质免疫印迹，我们在人睫状体或其他眼组织中未检测到hBest2。然而，经免疫沉淀富集后，在睫状体中鉴定出了hBest2，但在视网膜色素上皮中未检测到。利用免疫荧光，我们将hBest2定位在非色素上皮细胞的基底外侧质膜上。
结论
我们发现hBest2仅在非色素上皮细胞中的表达与小鼠相似。这些数据表明，Best2可能在人类房水流动和引流的调节中发挥功能作用。我们得出结论，Best2是青光眼治疗的一个新的潜在靶点。
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