Coupling switch of P2Y-IP₃ receptors mediates differential Ca²⁺ signaling in human embryonic stem cells and derived cardiovascular progenitor cells

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• 作者：Jijun Huang ; Min Zhang ; Peng Zhang ; He Liang ; Kunfu Ouyang…
• 关键词：P2Y receptors ; IP3 receptors ; Ca2+ signaling ; Pluripotent stem cells ; Lineage progenitors
• 刊名：Purinergic Signalling
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：12
• 期：3
• 页码：465-478
• 全文大小：5,087 KB
• 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
• 卷排序：12

文摘

Purinergic signaling mediated by P2 receptors (P2Rs) plays important roles in embryonic and stem cell development. However, how it mediates Ca2+ signals in human embryonic stem cells (hESCs) and derived cardiovascular progenitor cells (CVPCs) remains unclear. Here, we aimed to determine the role of P2Rs in mediating Ca2+ mobilizations of these cells. hESCs were induced to differentiate into CVPCs by our recently established methods. Gene expression of P2Rs and inositol 1,4,5-trisphosphate receptors (IP₃Rs) was analyzed by quantitative/RT-PCR. IP₃R3 knockdown (KD) or IP₃R2 knockout (KO) hESCs were established by shRNA- or TALEN-mediated gene manipulations, respectively. Confocal imaging revealed that Ca2+ responses in CVPCs to ATP and UTP were more sensitive and stronger than those in hESCs. Consistently, the gene expression levels of most P2YRs except P2Y₁ were increased in CVPCs. Suramin or PPADS blocked ATP-induced Ca2+ transients in hESCs but only partially inhibited those in CVPCs. Moreover, the P2Y₁ receptor-specific antagonist MRS2279 abolished most ATP-induced Ca2+ signals in hESCs but not in CVPCs. P2Y₁ receptor-specific agonist MRS2365 induced Ca2+ transients only in hESCs but not in CVPCs. Furthermore, IP₃R2KO but not IP₃R3KD decreased the proportion of hESCs responding to MRS2365. In contrast, both IP₃R2 and IP₃R3 contributed to UTP-induced Ca2+ responses while ATP-induced Ca2+ responses were more dependent on IP₃R2 in the CVPCs. In conclusion, a predominant role of P2Y₁ receptors in hESCs and a transition of P2Y-IP₃R coupling in derived CVPCs are responsible for the differential Ca2+ mobilization between these cells.