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Oxidative damage and mitochondrial functionality in hearts from KO UCP3 mice housed at thermoneutrality

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Abstract

The antioxidant role of mitochondrial uncoupling protein 3 (UCP3) is controversial. This work aimed to investigate the effects of UCP3 on the heart of mice housed at thermoneutral temperature, an experimental condition that avoids the effects of thermoregulation on mitochondrial activity and redox homeostasis, preventing the alterations related to these processes from confusing the results caused by the lack of UCP3. WT and KO UCP3 mice were acclimatized at 30 °C for 4 weeks and hearts were used to evaluate metabolic capacity and redox state. Tissue and mitochondrial respiration, the activities of the mitochondrial complexes, and the protein expression of mitochondrial complexes markers furnished information on mitochondrial functionality. The levels of lipid and protein oxidative damage markers, the activity of antioxidant enzymes, the reactive oxygen species levels, and the susceptibility to in vitro Fe-ascorbate-induced oxidative stress furnished information on redox state. UCP3 ablation reduced tissue and mitochondrial respiratory capacities, not affecting the mitochondrial content. In KO UCP3 mice, the mitochondrial complexes activities were lower than in WT without changes in their content. These effects were accompanied by an increase in the level of oxidative stress markers, ROS content, and in vitro susceptibility to oxidative stress, notwithstanding that the activities of antioxidant enzymes were not affected by UCP3 ablation. Such modifications are also associated with enhanced activation/phosphorylation of EIF2α, a marker of integrated stress response and endoplasmic reticulum stress (GRP778 BIP). The lack of UCP3 makes the heart more prone to oxidative insult by reducing oxygen consumption and increasing ROS. Our results demonstrate that UCP3 helps the cell to preserve mitochondrial function by mitigating oxidative stress.

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The datasets generated during and/or analyzed during the current study are available from the corresponding authors on reasonable request.

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Funding

This work was supported by grants from the University of Naples Federico II [Ricerca Dipartimentale 2018–2020].

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Authors and Affiliations

  1. Dipartimento di Scienze e Tecnologie, Università Degli Studi Di Napoli Parthenope, via Acton n. 38, -I-80133, Napoli, Italy

    Gaetana Napolitano

  2. Dipartimento di Biologia, Università di Napoli “Federico II,” Complesso Universitario Monte Sant’Angelo, Via Cinthia, 80126, Napoli, Italy

    Gianluca Fasciolo, Nunzia Magnacca, Assunta Lombardi & Paola Venditti

  3. Department of Sciences and Technologies, University of Sannio, Benevento, Italy

    Fernando Goglia

Authors
  1. Gaetana Napolitano
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  2. Gianluca Fasciolo
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  4. Fernando Goglia
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  5. Assunta Lombardi
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  6. Paola Venditti
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Contributions

N.G., L.A., G.F., and V.P. conceived and designed the experiments; F.G. and M.N. carried out all the animal studies and performed the experiments; G.N., G.F., L.A., and V.P. analyzed the data; G.F., L.A., and V.P. wrote the manuscript; all the authors revised the manuscript.

The authors declare that all data were generated in-house and that no paper mill was used.

Corresponding authors

Correspondence to Gaetana Napolitano, Assunta Lombardi or Paola Venditti.

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All animal protocols were approved by the Committee on the Ethics of Animal Experiments of the University of Naples Federico II and the Italian Minister of Health. Every effort was made to minimize animal pain and suffering.

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The authors declare no competing interests.

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Key points

• UCP3 ablation increases ROS content and reduces mitochondrial respiration

• KO UCP3 mice mitochondria show a high susceptibility to oxidative stress

• UCP3 regulates the ROS level in the heart of mice kept at thermoneutrality

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Napolitano, G., Fasciolo, G., Magnacca, N. et al. Oxidative damage and mitochondrial functionality in hearts from KO UCP3 mice housed at thermoneutrality. J Physiol Biochem 78, 415–425 (2022). https://doi.org/10.1007/s13105-022-00882-9

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  • DOI: https://doi.org/10.1007/s13105-022-00882-9

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