NEIBank项目中成人人类虹膜的表达序列标签分析：类固醇反应因子以及与视网膜色素上皮的相似性

Expressed sequence tag analysis of adult human iris for the NEIBank Project: steroid-response factors and similarities with retinal pigment epithelium.

作者信息

Wistow Graeme, Bernstein Steven L, Ray Sugata, Wyatt M Keith, Behal Amita, Touchman Jeffrey W, Bouffard Gerald, Smith Don, Peterson Katherine

机构信息

Section on Molecular Structure and Function, National Eye Institute, National Institutes of Health, Bethesda, MD 20892-2740, USA.

出版信息

Mol Vis. 2002 Jun 15;8:185-95.

PMID:12107412

Abstract

PURPOSE

The iris is a specialized tissue with important roles in the development and function of the eye. It is involved in diseases, including glaucoma and ocular melanoma, and its pigmented cells share an origin with the retinal pigment epithelium (RPE). Expressed sequence tag (EST) analysis of human iris has been performed to explore the repertoire of genes expressed in this tissue.

METHODS

An unamplified, un-normalized cDNA library (designated bx) was constructed from pooled (4-80 years old) human iris tissue. Over 2000 clones were picked and sequenced. Sequences were analyzed and clustered using GRIST (GRouping and Identification of Sequence Tags). The library was then normalized (and designated fg) and a further 2200 clones were sequenced for deeper examination of rarer sequence. Some sequences of interest were investigated further by standard methods.

RESULTS

From bx and fg libraries respectively, 1263 and 1604 clusters of expressed genes have been identified, giving a combined total of almost 2700 potentially unique genes. The most abundant novel transcript in bx is oculoglycan/opticin. Others include glucocorticoid induced leucine zipper protein (GILZ), Ris, a novel member of the Ras family, Iris Ring Finger (IRF), a member of the midline family, melastatin 2 (MLSN2), a member of the transient receptor potential calcium channel family, and iris expressed growth factor (IEGF), a member of the VEGF/PDGF family. Several factors involved in steroid responses are also represented.

CONCLUSIONS

The iris libraries are a rich source of novel as well as known genes, including molecular markers for pigmented cells that are also shared with RPE. A number of transcripts code for proteins involved in steroid response, with interesting implications for control of intraocular pressure. These sequence verified clones provide a nonredundant set for micro-array construction.

摘要

目的

虹膜是一种特殊组织，在眼睛的发育和功能中发挥着重要作用。它与包括青光眼和眼黑色素瘤在内的疾病有关，其色素细胞与视网膜色素上皮（RPE）有着共同的起源。已对人虹膜进行了表达序列标签（EST）分析，以探索该组织中表达的基因库。

方法

从汇集的（4至80岁）人虹膜组织构建了一个未扩增、未标准化的cDNA文库（命名为bx）。挑选了2000多个克隆进行测序。使用GRIST（序列标签的分组和鉴定）对序列进行分析和聚类。然后对文库进行标准化（命名为fg），并对另外2200个克隆进行测序，以更深入地检查稀有序列。通过标准方法对一些感兴趣的序列进行了进一步研究。

结果

分别从bx文库和fg文库中鉴定出1263个和1604个表达基因簇，总共给出了近2700个潜在的独特基因。bx中最丰富的新转录本是眼聚糖/视蛋白。其他包括糖皮质激素诱导的亮氨酸拉链蛋白（GILZ）、Ris（Ras家族的一个新成员）、虹膜环指蛋白（IRF，中线家族的一个成员）、褪黑素2（MLSN2，瞬时受体电位钙通道家族的一个成员）以及虹膜表达生长因子（IEGF，VEGF/PDGF家族的一个成员）。还发现了几种参与类固醇反应的因子。