COUP-TF1 在皮质锥体神经元分化的胚胎干细胞模型中调节甲状腺激素作用。
COUP-TF1 Modulates Thyroid Hormone Action in an Embryonic Stem-Cell Model of Cortical Pyramidal Neuronal Differentiation.
机构信息
1 Molecular Endocrinology Laboratory, VA Greater Los Angeles Healthcare System, Departments of Medicine and Physiology, David Geffen School of Medicine at UCLA , Los Angeles, California.
2 Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University , Shenyang, P. R. China .
出版信息
Thyroid. 2018 May;28(5):667-678. doi: 10.1089/thy.2017.0256. Epub 2018 Jan 10.
BACKGROUND
Thyroid hormone is critical for normal brain development and acts in a spatial and temporal specific pattern. Thyroid hormone excess, or deficiency, can lead to irreversible impairment of brain and sensory development. Chicken ovalbumin upstream-transcription factor 1 (COUP-TF1), expressed early in neuronal development, is essential to achieve normal brain structure. Thyroid hormone stimulation of gene expression is inversely correlated with the level of COUP-TF1 expression.
METHODS
An in vitro method of differentiating mouse embryonic stem (mES) cells into cortical neurons was utilized to study the influence of COUP-TF1 on thyroid hormone signaling in brain development. mES cells were cultured and differentiated in specific conditioned media, and a high percentage of nestin-positive progenitor neurons in the first stage, and cortical neurons in the second stage, was obtained with characteristic neuronal firing.
RESULTS
The number of nestin-positive progenitors, as determined by fluorescence-activated cell sorting analysis, was significantly greater with triiodothyronine (T3) treatment compared to control (p < 0.05). T3 enhanced the expression of cortical neuron marker (Tbr1 and Rc3) mRNAs. After COUP-TF1 knockdown, the number of nestin-positive progenitors was reduced compared to control (p < 0.05), but the number increased with T3 treatment. The mRNA of cortical neuronal gene markers was measured after COUP-TF1 knockdown. In the presence of T3, the peak expression of neuron markers Emx1, Tbr1, Camkiv, and Rc3 mRNA was earlier, at day 18 of differentiation, compared to control cells, at day 22. Furthermore, after COUP-TF1 knockdown, T3 induction of Rc3 and Tbr1 mRNA was significantly enhanced compared to cells expressing COUP-TF1.
CONCLUSION
These results indicate that COUP-TF1 plays an important role in modulating the timing and magnitude of T3-stimulated gene expression required for normal corticogenesis.
背景
甲状腺激素对正常脑发育至关重要，并具有时空特异性模式。甲状腺激素过多或不足会导致脑和感觉发育不可逆转的损害。鸡卵清蛋白上游转录因子 1（COUP-TF1）在神经元发育早期表达，对实现正常脑结构至关重要。甲状腺激素刺激基因表达与 COUP-TF1 表达水平呈负相关。
方法
利用体外分化小鼠胚胎干细胞（mES）为皮质神经元的方法，研究 COUP-TF1 对脑发育中甲状腺激素信号的影响。mES 细胞在特定条件培养基中培养和分化，第一阶段获得高比例的巢蛋白阳性祖细胞神经元，第二阶段获得具有特征性神经元放电的皮质神经元。
结果
通过荧光激活细胞分选分析确定，三碘甲状腺原氨酸（T3）处理组的巢蛋白阳性祖细胞数量明显多于对照组（p<0.05）。T3 增强了皮质神经元标志物（Tbr1 和 Rc3）mRNA 的表达。COUP-TF1 敲低后，与对照组相比，巢蛋白阳性祖细胞数量减少（p<0.05），但 T3 处理后数量增加。测量 COUP-TF1 敲低后皮质神经元基因标志物的 mRNA。在 T3 存在的情况下，与对照细胞相比，神经元标志物 Emx1、Tbr1、Camkiv 和 Rc3mRNA 的峰值表达更早，在分化的第 18 天，而不是第 22 天。此外，与表达 COUP-TF1 的细胞相比，COUP-TF1 敲低后，T3 诱导的 Rc3 和 Tbr1mRNA 的表达显著增强。
结论
这些结果表明，COUP-TF1 在调节 T3 刺激正常皮质发生所需的基因表达的时间和幅度方面发挥重要作用。
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