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

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Abstract

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.

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Acknowledgments

This work was supported by the grants from the National Natural Science of China Grants (Nos. 31030050 and 81520108004 to HTY, No. 31401167 to MZ); the Strategic Priority Research Program of CAS (No. XDA01020204 to HTY); the National Basic Research Program of China (No. 2014CB965100 to HTY); the National Science and Technology Major Project (No. 2012ZX09501001 to HTY); and the Major Program of Development Fund for Shanghai Zhangjiang National Innovation Demonstration Zone (No. ZJ2014-ZD-002 to HTY). We thank WiCell Research Institute for providing the H7 and H9 hESCs, and Dr. Heping Cheng (Peking University, Beijing, China) for providing the Flash Sniper software and for constructive discussion.

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Authors and Affiliations

  1. Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, Shanghai, 200031, China

    Jijun Huang, Min Zhang, Peng Zhang, He Liang & Huang-Tian Yang

  2. Second Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, 310009, China

    Jijun Huang & Huang-Tian Yang

  3. Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, 200120, China

    He Liang & Huang-Tian Yang

  4. Drug Discovery Center, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, 518055, China

    Kunfu Ouyang

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  1. Jijun Huang
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Correspondence to Huang-Tian Yang.

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Jijun Huang and Min Zhang are joint first co-authors.

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Huang, J., Zhang, M., Zhang, P. et al. Coupling switch of P2Y-IP3 receptors mediates differential Ca2+ signaling in human embryonic stem cells and derived cardiovascular progenitor cells. Purinergic Signalling 12, 465–478 (2016). https://doi.org/10.1007/s11302-016-9512-9

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