Generation of transgenic rats expressing green fluorescent protein in S-100β-producing pituitary folliculo-stellate cells and brain astrocytes
Generation of transgenic rats expressing green fluorescent protein in S-100β-producing pituitary folliculo-stellate cells and brain astrocytes
复制标题
DOI：
10.1210/en.2006-1390
复制
发表时间：
2007-04-01
期刊：
影响因子：
4.8
通讯作者：
Inoue, Kinji
中科院分区：
文献类型：
--
作者：
Itakura, Eisuke;Odaira, Kousuke;Inoue, Kinji
卵泡星状细胞（FS）被认为是前叶的支撑细胞或清道夫细胞。然而，FS细胞的确切功能和起源仍在讨论中。与脑星形胶质细胞一样，FS细胞含有S-100 β蛋白，固定后可用S-100 β蛋白抗体进行免疫细胞化学检测；但无法检测到活的FS细胞。在S-100 β蛋白基因启动子的控制下，产生表达绿色荧光蛋白（GFP）的转基因大鼠，可以检测FS活细胞，这可能是FS细胞研究的一个很好的工具。为了获得转基因大鼠，我们分析了S-100 β基因的启动子活性，发现内含子1对S-100 β基因的细胞特异性表达很重要。因此，我们获得了一个包含GFP基因的DNA构建体，位于S-100启动子的一部分，内含子1。我们将该构建体转染到大鼠胚胎中，成功地产生了转基因大鼠。转基因大鼠在前叶FS细胞中特异性表达GFP。GFP也在其他已知的S-100 β表达细胞中表达，如脑星形胶质细胞、脂肪细胞和软骨细胞。我们相信新获得的转基因大鼠将为FS细胞和其他S-100 β蛋白产生细胞的研究提供新的途径。
Folliculo-stellate (FS) cells are known to act as sustentacular cells or scavenger cells in the anterior lobe. However, the precise function and origin of FS cells are still under discussion. Like brain astrocytes, FS cells contain S-100 beta protein, and FS cells can be detected immunocytochemically using antibodies for S-100 beta protein after fixation; however, living FS cells can not be detected. The generation of transgenic rats expressing green fluorescent protein (GFP) under the control of S-100 beta protein gene promoter may allow the detection of living FS cells, which may be an excellent tool for the study of FS cells. With the aim of generation of transgenic rats, we analyzed the promoter activity of the S-100 beta gene and found that intron 1 is important for cell-specific expression of the S-100 beta gene. Therefore, we obtained a DNA construct containing GFP gene under a part of the S-100 promoter with intron 1. We transfected the construct into rat embryos and succeeded in generating transgenic rats. The transgenic rats expressed GFP in FS cells specifically in the anterior lobe. GFP is also expressed in other known S-100 beta-expressing cells, i.e. brain astrocytes, adipocytes, and chondrocytes. We believe that the newly generated transgenic rats will provide a new approach for the study of FS cells and other S-100 beta protein-producing cells.