TRANSGENIC INDICATOR MICE FOR STUDYING ACTIVATED RETINOIC ACID RECEPTORS DURING DEVELOPMENT
TRANSGENIC INDICATOR MICE FOR STUDYING ACTIVATED RETINOIC ACID RECEPTORS DURING DEVELOPMENT
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DOI：
10.1073/pnas.89.8.3347
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发表时间：
1992-04-15
影响因子：
11.1
通讯作者：
LINNEY, E
中科院分区：
文献类型：
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作者：
BALKAN, W;COLBERT, M;LINNEY, E
视黄酸 (RA) 受体 (RAR) 是配体诱导型转录因子，可与与 RA 调节基因的调节区域相关的特定 DNA 序列结合。由于 RA 被认为是正常发育和畸形学研究中的一个重要因素，因此人们希望拥有一种模型系统来检测发育过程中激活受体的存在。我们构建了一个重组报告基因，该基因具有三个拷贝的 RA 反应元件 (RARE)，从 RAR-beta-2 启动子 5' 到单纯疱疹病毒胸苷激酶启动子；该调节区与细菌β-半乳糖苷酶报告基因偶联。在 F9 胚胎癌细胞的瞬时转染测定中，该构建体可被 RA 诱导。具有该报告基因构建体的转基因胚胎在胚胎发生期间（从孕龄 7.5 天到 8.5 天之间开始）表现出受限且可重复的 β-半乳糖苷酶活性模式。第 8.5 天，在闭合的神经管和体节中检测到 β-半乳糖苷酶活性。来自提前 14 小时喂养 RA 的怀孕雌性的第 8.5 天胚胎表现出更大的 β-半乳糖苷酶活性强度和分布。同样，在妊娠后期，母体 RA 暴露导致胚胎 β-半乳糖苷酶表达增强。这种类型的转基因指示小鼠应该有助于详细说明激活的 RAR 在胚胎发育过程中的作用。
Retinoic acid (RA) receptors (RARs) are ligand-inducible transcription factors that bind to specific DNA sequences associated with the regulatory regions of RA-regulatable genes. Since RA has been implicated as an important factor both in normal development and in teratological studies, one would like to have a model system that detects the presence of activated receptors during development. We have constructed a recombinant reporter gene that has three copies of the RA response element (RARE) from the RAR-beta-2 promoter 5' to the herpes simplex virus thymidine kinase promoter; this regulatory region is coupled to the bacterial beta-galactosidase reporter gene. This construct was RA inducible in transient transfection assays in F9 embryonal carcinoma cells. Transgenic embryos with this reporter gene construct exhibited restricted and reproducible patterns of beta-galactosidase activity during embryogenesis, beginning between gestational ages day 7.5 and 8.5. At day 8.5, beta-galactosidase activity was detected in the closed neurotube and somites. Day 8.5 embryos, from pregnant females fed RA 14 hr earlier, exhibited a greater intensity and distribution of beta-galactosidase activity. Similarly, at later stages of gestation, maternal RA exposure resulted in enhanced embryonic beta-galactosidase expression. This type of transgenic indicator mouse should be useful in detailing the role of activated RARs during embryonic development.