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青鳉发育过程中脑型脂肪酸结合蛋白(fabp7)的表达

Expression of brain-type fatty acid-binding protein (fabp7) in medaka during development.

作者信息

Maruyama Kouichi, Kojima Ayako, Yasuda Takako, Suetomi Katsutoshi, Kubota Yoshihisa, Takahashi Sentaro, Ishikawa Yuji, Fujimori Akira

机构信息

Environmental Radiation Effects Research Group, Research Center for Radiation Protection, Japan.

出版信息

J Exp Zool B Mol Dev Evol. 2008 Nov 15;310(7):577-87. doi: 10.1002/jez.b.21226.

Abstract

Fatty acid-binding proteins (FABPs) belong to a multigene family of small intracellular proteins that bind hydrophobic ligands. Recent studies have indicated that FABP7 plays important roles in neurogenesis or neuronal migration in vertebrates. In this study, we isolated cDNA and the genomic fragment containing the fabp7 gene for medaka fish and examined the expression of the medaka fabp7 gene through the development of their central nervous system (CNS). The medaka fabp7 gene consists of four exons in approximately 1 kb of the genomic region. Its deduced amino acid sequence exhibits over 80% identity with those of other higher vertebrates. In situ hybridization analysis demonstrated that fabp7-positive cells first appear at stage 22 in a small dorsal domain of the retina, dorsal diencephalon, and rhombencephalon, then expand to the entire CNS including the retina and the spinal cord. In addition, we generated two lines of transgenic medaka with 1.7 kb upstream of the fabp7 gene combined with the enhanced-green fluorescence protein (EGFP) gene. The spatio-temporal expression patterns of EGFP in these animals were consistent with the results of in situ hybridization analysis. The result of our reporter assays with a series of truncated fabp7 promoters suggested that POU elements play a role in fabp7 expression in medaka as well as in other vertebrates. Our transgenic animal will contribute to clarifying the role of FABP7 in the development of CNS.

摘要

脂肪酸结合蛋白(FABPs)属于一类结合疏水性配体的细胞内小蛋白的多基因家族。最近的研究表明,FABP7在脊椎动物的神经发生或神经元迁移中起重要作用。在本研究中,我们分离了青鳉鱼的fabp7基因的cDNA和基因组片段,并通过其中枢神经系统(CNS)的发育来检测青鳉fabp7基因的表达。青鳉fabp7基因在约1 kb的基因组区域中由四个外显子组成。其推导的氨基酸序列与其他高等脊椎动物的氨基酸序列具有超过80%的同一性。原位杂交分析表明,fabp7阳性细胞首先出现在视网膜、背侧间脑和菱脑的一个小背侧区域的第22阶段,然后扩展到包括视网膜和脊髓在内的整个中枢神经系统。此外,我们构建了两系转基因青鳉,其fabp7基因上游1.7 kb与增强型绿色荧光蛋白(EGFP)基因结合。这些动物中EGFP的时空表达模式与原位杂交分析结果一致。我们用一系列截短的fabp7启动子进行的报告基因分析结果表明,POU元件在青鳉以及其他脊椎动物的fabp7表达中起作用。我们的转基因动物将有助于阐明FABP7在中枢神经系统发育中的作用。

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