Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.
Invest Ophthalmol Vis Sci. 2010 Mar;51(3):1738-46. doi: 10.1167/iovs.09-4520. Epub 2009 Oct 15.
The inner segments (IS) of the photoreceptors in vertebrates are enriched with polarity scaffold proteins, which maintain the integrity of many tissues by mediating cell-cell adhesion either directly or indirectly. The formation of photoreceptor mosaics may require differential adhesion among different types of photoreceptors. It is unknown whether any polarity proteins are selectively expressed in certain photoreceptors to mediate differential intercellular adhesion, which may be important for photoreceptor patterning. This study was undertaken to identify such polarity proteins.
To identify novel MAGUK-family (membrane-associated guanylate kinase) proteins that are similar to Nagie oko (Nok), the authors performed BLAST searches of the zebrafish genome with the Nok amino acid sequence as the query. The coding sequence of one of the identified genes was obtained and verified through RT-PCR and RACE (rapid amplification of cDNA ends). Its protein expression patterns were examined by immunomicroscopy and Western blot analysis. Morpholino knockdown technology was used for loss-of-function analyses.
The authors cloned a novel nok homolog and designated it photoreceptor-layer-nok-like (ponli). Unlike Nok, which is expressed broadly, Ponli is only expressed at the interface areas between the IS of the green, red, and blue cones in differentiated zebrafish retina.
Ponli is the first identified polarity protein that is not expressed in all types of photoreceptors. Ponli's selective distribution stimulates future investigations on its functions for photoreceptor mosaic formation.
脊椎动物光感受器的内节富含极性支架蛋白,这些蛋白通过直接或间接介导细胞-细胞黏附,维持许多组织的完整性。光感受器马赛克的形成可能需要不同类型的光感受器之间的差异黏附。目前尚不清楚是否有任何极性蛋白选择性地表达在特定的光感受器中,以介导细胞间的差异黏附,这对于光感受器的模式形成可能很重要。本研究旨在鉴定此类极性蛋白。
为了鉴定与 Nagie oko(Nok)相似的新型 MAGUK 家族(膜相关鸟苷酸激酶)蛋白,作者使用 Nok 氨基酸序列作为查询,对斑马鱼基因组进行 BLAST 搜索。通过 RT-PCR 和 RACE(快速扩增 cDNA 末端)获得并验证了其中一个鉴定基因的编码序列。通过免疫荧光显微镜和 Western blot 分析检查其蛋白表达模式。使用形态发生抑制技术进行功能丧失分析。
作者克隆了一个新型 nok 同源物,并将其命名为光感受器层-nok 样(ponli)。与广泛表达的 Nok 不同,ponli 仅在分化的斑马鱼视网膜中绿、红、蓝锥体的 IS 之间的界面区域表达。
ponli 是第一个被鉴定为不在所有类型的光感受器中表达的极性蛋白。ponli 的选择性分布激发了对其在光感受器马赛克形成中的功能的进一步研究。