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

Purinergic Signalling

Min Zhang²  Jijun Huang⁵  Huang-Tian Yang³  Kunfu Ouyang⁴  Peng Zhang²  He Liang¹ 

[1] Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine$$Tongji University School of Medicine$$;Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine$$;Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine$$Zhejiang University$$Tongji University School of Medicine$$;Peking University Shenzhen Graduate School$$;Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine$$Zhejiang University$$

关键词: Lineage progenitors;   

DOI  :  10.1007/s11302-016-9512-9

学科分类：分子生物学，细胞生物学和基因

来源: Springer

【 摘 要 】

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.

【 授权许可】

Unknown