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The Involvement of Cytochrome c Oxidase in Mitochondrial Fusion in Primary Cultures of Neonatal Rat Cardiomyocytes

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Abstract

Cytochrome c oxidase (CCO) is a copper-dependent enzyme of mitochondrial respiratory chain. In pressure overload-induced cardiac hypertrophy, copper level and CCO activity are both depressed, along with disturbance in mitochondrial fusion and fission dynamics. Copper repletion leads to recovery of CCO activity and normalized mitochondrial dynamics. The present study was undertaken to define the link between CCO activity and mitochondrial dynamic changes. Primary cultures of neonatal rat cardiomyocytes were treated with phenylephrine to induce cell hypertrophy. Hypertrophic cardiomyocytes were then treated with copper to reverse hypertrophy. In the hypertrophic cardiomyocytes, CCO activity was depressed and mitochondrial fusion was suppressed. Upon copper repletion, CCO activity was recovered and mitochondrial fusion was reestablished. Depression of CCO activity by siRNA targeting CCO assembly homolog 17 (COX17), a copper chaperone for CCO, led to fragmentation of mitochondria, which was not recoverable by copper supplementation. This study thus demonstrates that copper-dependent CCO is critical for mitochondrial fusion in the regression of cardiomyocyte hypertrophy.

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Fig. 1: Mitochondrial dynamic changes in hypertrophic cardiomyocytes
Fig. 2: The effect of Cu addition on PE-induced suppression of CCO activity
Fig. 3: Intracellular Cu contents in PE-treated cardiomyocytes and the effect of Cu supplementation
Fig. 4: The effects of COX17 deficiency on CCO activity and the subsequent effect on mitochondrial fragmentation
Fig. 5: Protein levels of MFN1 and DLP1 from COX17 siRNA -treated cells and the effect of Cu

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Acknowledgements

The authors want to thank Lin Bai, Suzheng Fan for providing technical assistance for mitochondrial imaging and AAS, respectively. This study was supported by National Science Foundation of China (Grant Number 81230004) and Sichuan University West China Hospital.

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  1. Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China

    Wen Yin, Rui Li, Xiaorong Feng & Y. James Kang

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  1. Wen Yin
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  2. Rui Li
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  3. Xiaorong Feng
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  4. Y. James Kang
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All authors participated in the design, analysis of the data, interpretation of the results, and review of the manuscript; WY, RL, and XRF carried out the experiments; YJK and WY wrote the manuscript.

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Correspondence to Y. James Kang.

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Yin, W., Li, R., Feng, X. et al. The Involvement of Cytochrome c Oxidase in Mitochondrial Fusion in Primary Cultures of Neonatal Rat Cardiomyocytes. Cardiovasc Toxicol 18, 365–373 (2018). https://doi.org/10.1007/s12012-018-9447-1

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