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视网膜色素上皮细胞间类视黄醇结合蛋白、视蛋白、细胞视黄醛结合蛋白和碱性成纤维细胞生长因子的差异表达

Differential expression of interphotoreceptor retinoid-binding protein, opsin, cellular retinaldehyde-binding protein, and basic fibroblastic growth factor.

作者信息

Gonzalez-Fernandez F, Van Niel E, Edmonds C, Beaver H, Nickerson J M, Garcia-Fernandez J M, Campohiaro P A, Foster R G

机构信息

Department of Ophthalmology, University of Virginia Health Sciences Center, Charlottesville 22908.

出版信息

Exp Eye Res. 1993 Apr;56(4):411-27. doi: 10.1006/exer.1993.1055.

Abstract

The mRNA for rat interphotoreceptor retinoid-binding protein (IRBP) consists of 5.2 kb and 6.4 kb transcripts which are expressed early during retinal development. Here, we characterized rat IRBP cDNA and genomic clones, determined the mechanism generating the two transcripts and compared their expression to mRNAs for opsin, cellular retinaldehyde-binding protein (CRAlBP) and basic fibroblast growth factor (bFGF). We found that human and rat IRBPs have a high degree of sequence homology (86% amino acid identity over the fourth repeat). RNA-PCR studies and Northern blot analysis, showed that the 6.4 kb mRNA has a longer 3'-untranslated region (UTR) than the 5.2 kb message. Both IRBP transcripts, but not the mRNA for opsin are present in the neonatal and adult pineal. During development, the mRNAs for IRBP and opsin reach one-half of their maximal levels by P5 and P11, respectively. The ratio of the two IRBP mRNAs remains constant throughout life. In contrast, the ratio of the longest to the shortest opsin mRNA decreases between E19 and 1 year of age. Immunohistochemistry demonstrated a marked increase in the amount of IRBP in the interphotoreceptor matrix between P1 and P9 corresponding to the enhanced expression of this mRNA. Up-regulation of opsin mRNA expression during the second postnatal week corresponds to the emergence of the outer segments. The temporal expression of CRAlBP is similar to IRBP while bFGF is not expressed until after photoreceptor differentiation is complete. In summary, two 3'-transcription termination sites explain the difference in IRBP mRNA sizes. The different temporal and tissue specific patterns of IRBP and opsin expression suggest that separate mechanisms control the expression of these two genes. The expression patterns of IRBP and CRAlBP are consistent with the role of vitamin A as a morphogen and bFGF in neuronal maintenance.

摘要

大鼠光感受器间类视黄醇结合蛋白(IRBP)的信使核糖核酸(mRNA)由5.2千碱基对(kb)和6.4 kb的转录本组成,它们在视网膜发育早期表达。在此,我们对大鼠IRBP的互补脱氧核糖核酸(cDNA)和基因组克隆进行了表征,确定了产生这两种转录本的机制,并将它们的表达与视蛋白、细胞视黄醛结合蛋白(CRAlBP)和碱性成纤维细胞生长因子(bFGF)的mRNA进行了比较。我们发现人和大鼠的IRBP具有高度的序列同源性(在第四个重复序列上氨基酸同一性为86%)。RNA聚合酶链反应(RNA-PCR)研究和Northern印迹分析表明,6.4 kb的mRNA比5.2 kb的信使核糖核酸具有更长的3'-非翻译区(UTR)。两种IRBP转录本在新生和成年松果体中均有表达,但视蛋白的mRNA则没有。在发育过程中,IRBP和视蛋白的mRNA分别在出生后第5天(P5)和第11天(P11)达到其最大水平的一半。两种IRBP mRNA的比例在整个生命过程中保持恒定。相比之下,最长和最短视蛋白mRNA的比例在胚胎第19天(E19)至1岁之间下降。免疫组织化学显示,在出生后第1天(P1)至第9天(P9)期间,光感受器间基质中IRBP的量显著增加,这与该mRNA表达的增强相对应。出生后第二周视蛋白mRNA表达的上调与外段的出现相对应。CRAlBP的时间表达与IRBP相似,而bFGF直到光感受器分化完成后才表达。总之,两个3'-转录终止位点解释了IRBP mRNA大小的差异。IRBP和视蛋白表达的不同时间和组织特异性模式表明,存在不同的机制控制这两个基因的表达。IRBP和CRAlBP的表达模式与视黄醇作为形态发生原以及bFGF在神经元维持中的作用一致。

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