Cardiomyogenesis of embryonic stem cells upon purinergic receptor activation by ADP and ATP

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• 作者：Safoura Mazrouei ; Fatemeh Sharifpanah ; Mohamed M. Bekhite…
• 关键词：Cardiomyocyte differentiation ; Purinergic receptors ; Intracellular [Ca2+]i signaling ; Embryonic stem cell ; ATP ; ADP
• 刊名：Purinergic Signalling
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：11
• 期：4
• 页码：491-506
• 全文大小：1,504 KB
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• 作者单位：Safoura Mazrouei (1)
  Fatemeh Sharifpanah (2)
  Mohamed M. Bekhite (1) (3)
  Hans-Reiner Figulla (1)
  Heinrich Sauer (2)
  Maria Wartenberg (1)
  
  1. University Heart Center, Clinic of Internal Medicine I, Friedrich Schiller University Jena, Jena, Germany
  2. Department of Physiology, Faculty of Medicine, Justus Liebig University, Giessen, Germany
  3. Department of Zoology, Faculty of Science, Tanta University, Tanta, Egypt
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Biomedicine
  Pharmacology and Toxicology
  Human Physiology
  Neurosciences
  Cancer Research
  
• 出版者：Springer Netherlands
• ISSN：1573-9546

文摘

Purinergic signaling may be involved in embryonic development of the heart. In the present study, the effects of purinergic receptor stimulation on cardiomyogenesis of mouse embryonic stem (ES) cells were investigated. ADP or ATP increased the number of cardiac clusters and cardiac cells, as well as beating frequency. Cardiac-specific genes showed enhanced expression of α-MHC, MLC2v, α-actinin, connexin 45 (Cx45), and HCN4, on both gene and protein levels upon ADP/ATP treatment, indicating increased cardiomyogenesis and pacemaker cell differentiation. Real-time RT-PCR analysis of purinergic receptor expression demonstrated presence of P2X1, P2X4, P2X6, P2X7, P2Y1, P2Y2, P2Y4, and P2Y6 on differentiating ES cells. ATP and ADP as well as the P2X agonists β,γ-methylenadenosine 5-triphosphate (β,γ-MetATP) and 8-bromoadenosine 5-triphosphate (8-Br-ATP) but not UTP or UDP transiently increased the intracellular calcium concentration ([Ca2+]_i) as evaluated by the calcium indicator Fluo-4, whereas no changes in membrane potential were observed. [Ca2+]_i transients induced by ADP/ATP were abolished by the phospholipase C-β (PLC-β) inhibitor U-73122, suggesting involvement of metabotropic P2Y receptors. Furthermore, partial inhibition of [Ca2+]_i transients was achieved in presence of MRS2179, a selective P2Y1 receptor antagonist, whereas PPADS, a non-selective P2 receptor inhibitor, completely abolished the [Ca2+]_i response. Consequently, cardiomyocyte differentiation was decreased upon long term co-incubation of cells with ADP and P2 receptor antagonists. In summary, activation of purinoceptors and the subsequent [Ca2+]_i transients enhance the differentiation of ES cells toward cardiomyocytes. Purinergic receptor stimulation may be a promising strategy to drive the fate of pluripotent ES cells into a particular population of cardiomyocytes. Keywords Cardiomyocyte differentiation Purinergic receptors Intracellular [Ca2+]_i signaling Embryonic stem cell ATP ADP