Identification of Small Signalling Molecules Promoting Cardiac-Specific Differentiation of Mouse Embryonic Stem Cells
Identification of Small Signalling Molecules Promoting Cardiac-Specific Differentiation of Mouse Embryonic Stem Cells
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DOI：
10.1159/000097608
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发表时间：
2006-01
影响因子：
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通讯作者：
A. Sachinidis;S. Schwengberg;R. Hippler-Altenburg;D. Mariappan;Naidu Kamisetti;B. Seelig;A. Berkessel;J. Hescheler
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文献类型：
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作者：
A. Sachinidis;S. Schwengberg;R. Hippler-Altenburg;D. Mariappan;Naidu Kamisetti;B. Seelig;A. Berkessel;J. Hescheler
识别参与心肌发生的信号级联对于优化体外胚胎干细胞（ES细胞）生成心肌细胞至关重要。我们使用在α-肌球蛋白重链（α-MHC）启动子控制下表达增强型绿色荧光蛋白（EGFP）的转基因ES细胞系（pαMHC-EGFP）来研究33种干扰几种信号级联的小分子对心肌发生的影响。有趣的是，L型钙通道阻滞剂维拉帕米以及蛋白磷酸酶2B抑制剂环孢菌素发挥了最显著的促心肌生成作用。Forskolin（腺苷酸环化酶刺激剂）发挥最显着的抗心肌作用。与发育晚期胚状体（embryoid bodies，EBs）相比，维拉帕米或环孢菌素刺激早期胚状体（1日龄）48 h可产生较强的心肌诱导作用。刺激早期发育阶段的EB 48 h后，α-MHC mRNA和EGFP mRNA的表达增强。没有表达？已经观察到平滑肌肌动蛋白或血小板内皮细胞粘附分子-1（PECM-1）以及神经元基因（巢蛋白，神经丝H），证明这两种分子优先具有促心肌发生作用。
Identification of signalling cascades involved in cardiomyogenesis is crucial for optimising the generation of cardiomyocytes from embryonic stem cells (ES cells) in vitro. We used a transgenic ES cell lineage expressing enhanced green fluorescent protein (EGFP) under the control of the α-myosin heavy chain (α-MHC) promoter (pαMHC-EGFP) to investigate the effects of 33 small molecules interfering with several signalling cascades on cardiomyogenesis. Interestingly, the L-Type Ca2+ channel blocker Verapamil as well as Cyclosporin, an inhibitor of the protein phosphatase 2B, exerted the most striking pro-cardiomyogenic effect. Forskolin (adenylate cyclase stimulator) exerted the most striking anti-cardiomyogenic effect. The cardiomyogenic effect of Cyclosporin and Verapamil correlated with an expression of early cardiac markers Nkx2.5 and GATA4.Compared to the effects on late developmental stage embryoid bodies (EBs) stimulation of early developmental stage EBs (1-day old) with Verapamil or Cyclosporin for 48 h resulted in a potent cardiomyogenic effect. Accordingly, enhanced expression of α-MHC mRNA and EGFP mRNA was observed after stimulation of the early developmental stage EBs for 48 h. No expression of ?-smooth muscle actin or platelet endothelial cell adhesion molecule-1 (PECM-1) as well as of neuronal genes (Nestin, Neurofilament H) has been observed demonstrating a preferentially pro-cardiomyogenic effect by both molecules.