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Cardiac Na+-Ca2+ exchanger 1 (ncx1h) is critical for the ventricular cardiomyocyte formation via regulating the expression levels of gata4 and hand2 in zebrafish

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Abstract

Ca2+ signaling is critical for heart development; however, the precise roles and regulatory pathways of Ca2+ transport proteins in cardiogenesis remain largely unknown. Sodium-calcium exchanger 1 (Ncx1) is responsible for Ca2+ efflux in cardiomyocytes. It is involved in cardiogenesis, while the mechanism is unclear. Here, using the forward genetic screening in zebrafish, we identified a novel mutation at a highly-conserved leucine residue in ncx1 gene (mutantLDD353/ncx1hL154P) that led to smaller hearts with reduced heart rate and weak contraction. Mechanistically, the number of ventricular but not atrial cardiomyocytes was reduced in ncx1hL154P zebrafish. These defects were mimicked by knockdown or knockout of ncx1h. Moreover, ncx1hL154P had cytosolic and mitochondrial Ca2+ overloading and Ca2+ transient suppression in cardiomyocytes. Furthermore, ncx1hL154P and ncx1h morphants downregulated cardiac transcription factors hand2 and gata4 in the cardiac regions, while overexpression of hand2 and gata4 partially rescued cardiac defects including the number of ventricular myocytes. These findings demonstrate an essential role of the novel 154th leucine residue in the maintenance of Ncx1 function in zebrafish, and reveal previous unrecognized critical roles of the 154th leucine residue and Ncx1 in the formation of ventricular cardiomyocytes by at least partially regulating the expression levels of gata4 and hand2.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (81520108004, 81470422), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16010201), and National Key R&D Program of China (2017YFA 0103700, 2016YFC1301204) to H.-T.Y., and Shanghai Natural Science Foundation (17ZR1435500) to J.H. We acknowledge Dr. Atsushi Miyawaki and Dr. Akiko Ishioka for providing the two Ca2+ indicator transgenic zebrafish lines, and Prof. Tao Zhong for providing the vmhc-EGFP plasmid. We also thank Min Den, Yi Jin, Mei Dong, and Yukun Jiang for technical assistance.

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Authors and Affiliations

  1. CAS Key Laboratory of Tissue Microenvironment and Tumor, Laboratory of Molecular Cardiology and Laboratory of Development and Diseases, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences (CAS), CAS, Shanghai, 200031, China

    Liming Chu, Huimin Yin, Lei Gao, Yu Xia, Chiyuan Zhang, Tingxi Liu, Jijun Huang, Yong Zhou & Huang-Tian Yang

  2. Institute for Stem Cell and Regeneration, CAS, Beijing, 100101, China

    Liming Chu, Huimin Yin, Lei Gao, Yu Xia, Chiyuan Zhang, Tingxi Liu, Jijun Huang, Yong Zhou & Huang-Tian Yang

  3. State Key Laboratory for Medical Genomics, Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

    Li Gao & Yi Chen

  4. Department of Biomedical Engineering, The Johns Hopkins University, Baltimore, MD, 21205, USA

    Kenneth R. Boheler

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  1. Liming Chu
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  2. Huimin Yin
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Correspondence to Yong Zhou or Huang-Tian Yang.

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Cardiac Na+-Ca2+ exchanger 1 (ncx1h) is critical for the ventricular cardiomyocyte formation via regulating the expression levels of gata4 and hand2 in zebrafish

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Chu, L., Yin, H., Gao, L. et al. Cardiac Na+-Ca2+ exchanger 1 (ncx1h) is critical for the ventricular cardiomyocyte formation via regulating the expression levels of gata4 and hand2 in zebrafish. Sci. China Life Sci. 64, 255–268 (2021). https://doi.org/10.1007/s11427-019-1706-1

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