Lutz B, Kuratani S, Cooney A J, Wawersik S, Tsai S Y, Eichele G, Tsai M J
Department of Biochemistry, Baylor College of Medicine, Houston, TX 77030.
Development. 1994 Jan;120(1):25-36. doi: 10.1242/dev.120.1.25.
Members of the steroid/thyroid hormone receptor superfamily are involved in the control of cell identity and of pattern formation during embryonic development. Chicken ovalbumin upstream promoter-transcription factors (COUP-TFs) can act as regulators of various steroid/thyroid hormone receptor pathways. To begin to study the role of COUP-TFs during embryogenesis, we cloned a chicken COUP-TF (cCOUP-TF II) which is highly homologous to human COUP-TF II. Northern analysis revealed high levels of cCOUP-TF II transcripts during organogenesis. Nuclear extracts from whole embryos and from embryonic spinal cords were used in electrophoretic mobility shift assays. These assays showed that COUP-TF protein is present in these tissues and is capable of binding to a COUP element (a direct repeat of AGGTCA with one base pair spacing). Analysis of cCOUP-TF expression by in situ hybridization revealed high levels of cCOUP-TF II mRNA in the developing spinal motor neurons. Since the ventral properties of the spinal cord, including the development of motor neurons, is in part established by inductive signals from the notochord, we transplanted an additional notochord next to the dorsal region of the neural tube in order to induce ectopic motor neurons. We observed that an ectopic notochord induced cCOUP-TF II gene expression in the dorsal spinal cord in a region coextensive with ectopic domains of SC1 and Islet-1, two previously identified motor neuron markers. Collectively, our studies raise the possibility that cCOUP-TF II is involved in motor neuron development.
类固醇/甲状腺激素受体超家族的成员参与胚胎发育过程中细胞特性的控制和模式形成。鸡卵清蛋白上游启动子转录因子(COUP-TFs)可作为各种类固醇/甲状腺激素受体途径的调节因子。为了开始研究COUP-TFs在胚胎发生过程中的作用,我们克隆了一种与人类COUP-TF II高度同源的鸡COUP-TF(cCOUP-TF II)。Northern分析显示在器官发生过程中cCOUP-TF II转录本水平很高。来自全胚胎和胚胎脊髓的核提取物用于电泳迁移率变动分析。这些分析表明COUP-TF蛋白存在于这些组织中,并且能够与COUP元件(AGGTCA的直接重复序列,间隔一个碱基对)结合。通过原位杂交分析cCOUP-TF的表达,发现在发育中的脊髓运动神经元中cCOUP-TF II mRNA水平很高。由于脊髓的腹侧特性,包括运动神经元的发育,部分是由来自脊索的诱导信号建立的,我们在神经管背侧区域旁边移植了一个额外的脊索,以诱导异位运动神经元。我们观察到异位脊索在背侧脊髓中诱导了cCOUP-TF II基因表达,该区域与SC1和Islet-1这两个先前确定的运动神经元标记的异位区域共延。总的来说,我们的研究提出了cCOUP-TF II参与运动神经元发育的可能性。