Endothelin-induced differentiation of Nkx2.5+ cardiac progenitor cells into pacemaking cells

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The mechanisms governing the development of cardiac pacemaking and conduction system are not well understood. In order to provide evidence for the derivation of pacemaking cells and the signal that induce and maintain the cells in the developing heart, Nkx2.5+ cardiac progenitor cells (CPCs) were isolated from embryonic heart tubes of rats. Endothelin-1 was subsequently added to the CPCs to induce differentiation of them towards cardiac pacemaking cells. After the treatment, Nkx2.5+ CPCs displayed spontaneous beating and spontaneously electrical activity as what we have previously described. Furthermore, RT-PCR and immunofluorescence staining demonstrated that Tbx3 expression was increased and Nkx2.5 expression was decreased in the induced cells 4 days after ET-1 treatment. And the significantly increased expression of Hcn4 and connexin-45 were detected in the induced cells 10 days after the treatment. In addition, Nkx2.5+ CPCs were transfected with pGCsi-Tbx3 4 days after ET-1 treatment in an attempt to determine the transcription regulatory factor governing the differentiation of the cells into cardiac pacemaking cells. The results showed that silencing of Tbx3 decreased the pacemaking activity and led to down-regulation of pacemaker genes in the induced cells. These results confirmed that Nkx2.5+ CPCs differentiated into cardiac pacemaking cells after being treated with ET-1 and suggested that an ET-1-Tbx3 molecular pathway govern/mediate this process. In conclusion, our study support the notion that pacemaking cells originate from Nkx2.5+ CPCs present in embryonic heart tubes and endothelin-1 might be involved in diversification of cardiomyogenic progenitors toward the cells.

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This work was supported by the National Natural Science Foundation of China (No. 30772158, No. 30970737, No. 81071603) and the Shanghai Natural Science Foundation of China (No. 07ZR14140, No. 09JC1417700, and No. 09ZR1439300).

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Correspondence to Chuan-Sen Zhang or Xiang-Qun Yang.

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Xi Zhang and Jin-Ping Guo contributed equally to this work and should be considered as co-first authors.

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Supplementary material 1 (MOV 1810 kb)

Supplementary material 2 (MOV 1704 kb)

Supplementary material 3 (MOV 1685 kb)

Supplementary material 4 (MOV 1929 kb)

Supplementary material 5 (MOV 1867 kb)

Supplementary material 6 (MOV 2039 kb)

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Zhang, X., Guo, J., Chi, Y. et al. Endothelin-induced differentiation of Nkx2.5+ cardiac progenitor cells into pacemaking cells. Mol Cell Biochem 366, 309–318 (2012) doi:10.1007/s11010-012-1309-8

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  • Cardiac progenitor cell
  • Cardiac pacemaking cell
  • Embryonic heart tube
  • Cardiac development