Ahmed Farid, Torrado Mario, Zinovieva Rina D, Senatorov Vladimir V, Wistow Graeme, Tomarev Stanislav I
Section of Molecular Mechanisms of Glaucoma, Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, MD 20814-9692, USA.
Invest Ophthalmol Vis Sci. 2004 Sep;45(9):3081-90. doi: 10.1167/iovs.04-0302.
To characterize gene expression pattern in the combined tissues of the rat iridocorneal angle by expressed sequence tag (EST) analysis, as part of the NEIBank project.
RNA was extracted from dissected tissues of the rat iridocorneal angle (iris, ciliary body, trabecular meshwork, and Schlemm's canal) and used to construct unamplified, non-normalized cDNA libraries in the pSPORT1 vector. Approximately 5000 clones were sequenced from the 5'-end. Clones were clustered and identified using the GRIST software, a procedure based on BLAST comparisons. Complete sequences of several novel cDNAs showing eye-preferred expression patterns were obtained. The expression patterns of several genes have been investigated by Northern blot and in situ hybridization, as well as by RT-PCR.
After analysis and removal of non-mRNA sequences, 2195 independent clusters, potentially representing individual eye angle-expressed clones were obtained. The expression profile of the combined rat eye angle tissues was more similar to that of the human iris than to human trabecular meshwork. Several cDNAs encoding transcription factors essential for normal eye development and function including Pax-6, Six3, c-Maf, Maf1, Sox-4, Foxc1, Rx, and Ldb2 were present among sequenced clones. A number of tested cDNAs showed eye-preferred expression patterns. Myocilin, which is abundant in human eye angle tissues, was not observed in the rat collection; however, transcripts for three other olfactomedin-domain proteins were seen. Latrotoxin receptor (CL1AA) and optimedin were shown to be expressed in the iris and ciliary body, as well as in the ganglion and inner nuclear cell layers of the retina, whereas the rat orthologue of the human HNOEL-iso gene was expressed in the iris and sclera and less actively in the trabecular meshwork, retina, and optic nerve.
The iridocorneal libraries are a good source of novel uncharacterized genes and molecular markers for the tissues of the eye angle. Although myocilin is not abundantly expressed in rat eye angle, other olfactomedin-containing genes are expressed there and may play important roles in normal eye function and disease.
作为NEIBank项目的一部分，通过表达序列标签（EST）分析来描述大鼠虹膜角膜角联合组织中的基因表达模式。
从大鼠虹膜角膜角（虹膜、睫状体、小梁网和施莱姆管）的解剖组织中提取RNA，并用于构建pSPORT1载体中的未扩增、非标准化cDNA文库。从5'端对大约5000个克隆进行测序。使用GRIST软件（一种基于BLAST比较的程序）对克隆进行聚类和鉴定。获得了几个显示出眼部优先表达模式的新cDNA的完整序列。通过Northern印迹、原位杂交以及RT-PCR研究了几个基因的表达模式。
在分析并去除非mRNA序列后，获得了2195个独立的聚类，可能代表单个眼角表达的克隆。大鼠眼角联合组织的表达谱与人类虹膜的表达谱比与人类小梁网的表达谱更相似。在测序的克隆中存在几个编码对正常眼睛发育和功能至关重要的转录因子的cDNA，包括Pax-6、Six3、c-Maf、Maf1、Sox-4、Foxc1、Rx和Ldb2。许多测试的cDNA显示出眼部优先表达模式。在人眼角组织中丰富的肌纤蛋白在大鼠样本中未观察到；然而，发现了另外三种嗅觉介质结构域蛋白的转录本。Latrodoxin受体（CL1AA）和视优明在虹膜和睫状体以及视网膜的神经节和内核细胞层中表达，而人类HNOEL-iso基因的大鼠同源物在虹膜和巩膜中表达，在小梁网、视网膜和视神经中的表达活性较低。
虹膜角膜文库是眼角组织中新型未表征基因和分子标记的良好来源。虽然肌纤蛋白在大鼠眼角中不大量表达，但其他含嗅觉介质的基因在那里表达，可能在正常眼睛功能和疾病中起重要作用。
