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AP-2α和AP-2β在发育中的鸡视网膜中的差异表达:AP-2对R-FABP启动子活性的抑制作用

Differential expression of AP-2alpha and AP-2beta in the developing chick retina: repression of R-FABP promoter activity by AP-2.

作者信息

Bisgrove D A, Godbout R

机构信息

Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, Canada.

出版信息

Dev Dyn. 1999 Mar;214(3):195-206. doi: 10.1002/(SICI)1097-0177(199903)214:3<195::AID-AJA3>3.0.CO;2-8.

Abstract

Retinal fatty acid binding protein (R-FABP) is the avian counterpart of murine brain FABP implicated in glial cell differentiation and neuronal cell migration. R-FABP is highly expressed in the undifferentiated retina and brain of chick embryos. We have previously shown by in vitro studies that the AP-2 transcription factor binds to a consensus AP-2 binding site in the R-FABP promoter region. Based on the expression pattern of AP-2 in the developing retina and on mutational analysis of the AP-2 binding site in DNA transfection experiments, we proposed that AP-2 could be involved in the down-regulation of R-FABP transcription. Here, we describe the cDNA isolation of two members of the AP-2 family expressed in the chick retina, AP-2alpha and AP-2beta. We show that R-FABP mRNA and the AP-2 factors are expressed in mutually exclusive patterns in the differentiating retina: whereas AP-2alpha and AP-2beta are selectively expressed either in amacrine, or in amacrine and horizontal cells, respectively, R-FABP mRNAis found in Muller glial cells and/or bipolar cells. Furthermore, a decrease in R-FABP-dependent expression is obtained upon cotransfection of primary retinal cultures with AP-2 expression vectors and a CAT reporter construct. The early and cell-specific expression of AP-2alpha and AP-2beta in the developing retina suggest a role for this transcription factor family in the early steps of amacrine and horizontal cell differentiation. Repression of the R-FABP gene in these cells may be an important component of their developmental program.

摘要

视网膜脂肪酸结合蛋白(R-FABP)是与神经胶质细胞分化和神经元细胞迁移有关的小鼠脑脂肪酸结合蛋白的禽类对应物。R-FABP在鸡胚未分化的视网膜和脑中高度表达。我们之前通过体外研究表明,AP-2转录因子与R-FABP启动子区域的一个共有AP-2结合位点结合。基于AP-2在发育中的视网膜中的表达模式以及DNA转染实验中AP-2结合位点的突变分析,我们提出AP-2可能参与R-FABP转录的下调。在这里,我们描述了在鸡视网膜中表达的AP-2家族两个成员AP-2α和AP-2β的cDNA分离。我们表明,在分化的视网膜中,R-FABP mRNA和AP-2因子以相互排斥的模式表达:AP-2α和AP-2β分别选择性地在无长突细胞或无长突细胞和水平细胞中表达,而R-FABP mRNA则在穆勒胶质细胞和/或双极细胞中发现。此外,用AP-2表达载体和CAT报告构建体共转染原代视网膜培养物后,R-FABP依赖性表达降低。AP-2α和AP-2β在发育中的视网膜中的早期和细胞特异性表达表明该转录因子家族在无长突细胞和水平细胞分化的早期步骤中发挥作用。在这些细胞中抑制R-FABP基因可能是其发育程序的一个重要组成部分。

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