Functional notch signaling is required for BMP4-induced inhibition of myogenic differentiation
Functional notch signaling is required for BMP4-induced inhibition of myogenic differentiation
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DOI：
10.1242/dev.00834
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发表时间：
2003-12-01
期刊：
影响因子：
4.6
通讯作者：
Lendahl, U
中科院分区：
文献类型：
--
作者：
Dahlqvist, C;Blokzijl, A;Lendahl, U
骨形态发生蛋白（BMP）和Notch信号通路对细胞分化至关重要。在许多情况下，这两种途径的作用相似;例如，抑制肌源性分化。目前尚不清楚这种抑制是否是由不同的机制或Notch和BMP信号之间的相互作用引起的。在这里，我们证明了功能性Notch信号传导是BMP 4介导的肌肉干细胞分化阻滞所必需的，即卫星细胞和肌源性细胞系C2 C12。在诱导分化过程中加入BMP 4显著减少了分化的卫星细胞和C2 C12细胞的数量。通过使用γ-分泌酶抑制剂L-685，458阻断Notch信号传导或通过引入Notch信号介体CSL的显性阴性形式，在BMP 4处理的培养物中基本上恢复了分化。向C2 C12细胞中添加BMP 4增加了两个立即Notch应答基因Hes 1和Hey 1的转录，这种作用被L-685，458废除。通过Notch 1胞内结构域（Notch 1 ICD）和BMP 4协同激活3 kb Hey 1启动子报告基因构建体。BMP 4介体SMAD 1模拟了Hey 1启动子的BMP激活。一个合成的Notch响应启动子不含SMAD 1结合位点，SMAD 1响应，表明SMAD 1的DNA结合活性是不需要激活。因此，Notch 1 ICD和SMAD 1在体外结合实验中相互作用。因此，本文提供的数据为Notch和BMP信号传导途径之间的直接相互作用提供了证据，并表明Notch在BMP介导的分化控制的某些方面的执行中具有关键作用。
The bone morphogenetic protein (BMP) and Notch signaling pathways are crucial for cellular differentiation. In many cases, the two pathways act similarly; for example, to inhibit myogenic differentiation. It is not known whether this inhibition is caused by distinct mechanisms or by an interplay between Notch and BMP signaling. Here we demonstrate that functional Notch signaling is required for BMP4-mediated block of differentiation of muscle stem cells, i.e. satellite cells and the myogenic cell line C2C12. Addition of BMP4 during induction of differentiation dramatically reduced the number of differentiated satellite and C2C12 cells. Differentiation was substantially restored in BMP4-treated cultures by blocking Notch signaling using either the gamma-secretase inhibitor L-685,458 or by introduction of a dominant-negative version of the Notch signal mediator CSL. BMP4 addition to C2C12 cells increased transcription of two immediate Notch responsive genes, Hes1 and Hey1, an effect that was abrogated by L-685,458. A 3 kb Hey1-promoter reporter construct was synergistically activated by the Notch 1 intracellular domain (Notch 1 ICD) and BMP4. The BMP4 mediator SMAD1 mimicked BMP activation of the Hey1 promoter. A synthetic Notch-responsive promoter containing no SMAD1 binding sites responded to SMAD1, indicating that DNA-binding activity of SMAD1 is not required for activation. Accordingly, Notch 1 ICD and SMAD1 interacted in binding experiments in vitro. Thus, the data presented here provide evidence for a direct interaction between the Notch and BMP signaling pathways, and indicate that Notch has a crucial role in the execution of certain aspects of BMP-mediated differentiation control.