Fibroblast growth factor receptor-1 is essential for in vitro cardiomyocyte development
Fibroblast growth factor receptor-1 is essential for in vitro cardiomyocyte development
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DOI：
10.1161/01.res.0000089460.12061.e1
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发表时间：
2003-09-05
影响因子：
20.1
通讯作者：
Presta, M
中科院分区：
文献类型：
--
作者：
Dell'Era, P;Ronca, R;Presta, M
成纤维细胞生长因子（FGF）/FGF受体（FGFR）信号传导在中胚层形成和模式化中起关键作用。在果蝇中进行的无情突变体研究表明，在不同的FGFR中，FGFR 1可能在心脏发生中发挥作用。然而，fgfr 1（-/-）小鼠在心脏形成前的原肠胚形成期间死亡。为了确定FGFR 1在心脏发育中的作用，我们研究了小鼠fgfr 1（-/-）和fgfr 1（-/-）胚胎干细胞（ES）在体外分化为心肌细胞的能力。在分化的第9至10天，在>90%的源自fgfr 1（+/-）ES细胞的三维胚状体（EBs）中观察到搏动的心肌细胞簇。相比之下，10%或更少的fgfr 1（-/-）EB在第16天显示搏动灶。因此，fgfr 1（-/-）EB的特征是早期心脏转录因子Nkx2.5和d-Hand以及晚期结构心脏基因肌球蛋白重链（MHC）-α、MHC-β和心室肌球蛋白轻链的表达受损。纯合fgfr 1突变也导致中胚层相关早期基因表达的改变，包括nodal、BMP 2、BMP 4、T（bra）和sonic hedgehog。然而，fgfr 1（-/-）和fgfr 1（-/-）EB类似地表达心源性前体、内皮、造血和骨骼肌标志物，表明fgfr 1无效突变在分化的ES细胞中对心肌细胞发育产生选择性影响。因此，FGFR信号传导的抑制剂，包括FGFR 1酪氨酸激酶抑制剂SU 5402、MEK 1/2抑制剂U 0126和蛋白激酶C抑制剂GF 109，都阻止了fgfr 1（-/-）EB中的心肌细胞分化，而不影响造血/内皮标志物flk-1的表达。总之，数据表明FGFR 1介导的信号在心肌细胞发育中的作用是非冗余的。
Fibroblast growth factor (FGF)/FGF receptor ( FGFR) signaling plays a crucial role in mesoderm formation and patterning. Heartless mutant studies in Drosophila suggest that FGFR1, among the different FGFRs, may play a role in cardiogenesis. However, fgfr1(-/-) mice die during gastrulation before heart formation. To establish the contribution of FGFR1 in cardiac development, we investigated the capacity of murine fgfr1(-/-) and fgfr1(-/-) embryonic stem (ES) cells to differentiate to cardiomyocytes in vitro. Clusters of pulsating cardiomyocytes were observed in >90% of 3-dimensional embryoid bodies (EBs) originated from fgfr1(+/-) ES cells at day 9 to 10 of differentiation. In contrast, 10% or less of fgfr1(-/-) EBs showed beating foci at day 16. Accordingly, fgfr1(-/-) EBs were characterized by impaired expression of early cardiac transcription factors Nkx2.5 and d-Hand and of late structural cardiac genes myosin heavy chain (MHC)-alpha, MHC-beta, and ventricular myosin light chain. Homozygous fgfr1 mutation resulted also in alterations of the expression of mesoderm-related early genes, including nodal, BMP2, BMP4, T( bra), and sonic hedgehog. Nevertheless, fgfr1(-/-) and fgfr1(-/-) EBs similarly express cardiogenic precursor, endothelial, hematopoietic, and skeletal muscle markers, indicating that fgfr1-null mutation exerts a selective effect on cardiomyocyte development in differentiating ES cells. Accordingly, inhibitors of FGFR signaling, including the FGFR1 tyrosine kinase inhibitor SU 5402, the MEK1/2 inhibitor U0126, and the protein kinase C inhibitor GF109 all prevented cardiomyocyte differentiation in fgfr1(-/-) EBs without affecting the expression of the hematopoietic/ endothelial marker flk-1. In conclusion, the data point to a nonredundant role for FGFR1-mediated signaling in cardiomyocyte development.