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Dissociation of JNK Activation from Elevated Levels of Reactive Oxygen Species, Cytochrome c Release, and Cell Death in NGF-Deprived Sympathetic Neurons

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Abstract

Withdrawal of nerve growth factor (NGF) from sympathetic neurons causes their apoptotic death. Activation of c-Jun NH2-terminal kinase (JNK) may contribute to this death by the induction and phosphorylation of pro-apoptotic Bcl-2 proteins, such as Bax, that are involved in cytochrome c release from mitochondria and reactive oxygen species (ROS) production. Induction of either JNK or ROS may stimulate the other, and both may regulate release of apoptogenic factors from the mitochondria. In order to discern the relationship between JNK and ROS in apoptosis, we treated NGF-deprived, mouse sympathetic neurons with a JNK inhibitor and examined the effect on several important apoptotic events. Block of JNK activation prevented induction of c-Jun expression and resulted in a dose-dependent, yet surprisingly modest, increase in cell survival after 48 h of NGF deprivation. JNK suppression was also not sufficient to prevent the elevation in ROS or the release of cytochrome c from the mitochondria in NGF-deprived sympathetic neurons. Bax deletion prevents apoptotic death of NGF-deprived neurons by preventing release of cytochrome c from their mitochondria. It also prevents increased ROS on NGF deprivation. However, we found that induction of c-Jun in cells lacking Bax was equivalent to that in wild-type neurons. Our results suggest that while JNK activation plays an important role in many forms of apoptosis, it may not be a crucial regulator of Bax-dependent events involved in the apoptotic death of mouse sympathetic neurons deprived of NGF and that ROS is not involved in its activation in these cells.

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Acknowledgements

This study was supported by National Institutes of Health grant R03 AG051205.

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Authors and Affiliations

  1. Center of Mitochondrial and Epigenomic Medicine, Children’s Hospital of Philadelphia’s Colket Translational Research Building, 3501 Civic Center Blvd, Room 6100, Philadelphia, PA, 19104, USA

    Meagan J. McManus

  2. Department of Pharmaceutical and Biomedical Sciences, The University of Georgia College of Pharmacy, 357 Wilson Pharmacy, Athens, GA, 30602, USA

    James L. Franklin

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Correspondence to James L. Franklin.

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This work was approved by the Institutional Animal Care and Use Committee of the University of Georgia (animal use protocol no. A2014 03-005-Y3-A2). The university animal care and use program is accredited by Association for Assessment and Accreditation of Laboratory Animal Care ( AAALAC International ), is licensed by the USDA, and maintains an Assurance of Compliance with the Public Health Service.

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The authors declare that they have no conflicts of interest.

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McManus, M.J., Franklin, J.L. Dissociation of JNK Activation from Elevated Levels of Reactive Oxygen Species, Cytochrome c Release, and Cell Death in NGF-Deprived Sympathetic Neurons. Mol Neurobiol 55, 382–389 (2018). https://doi.org/10.1007/s12035-016-0332-2

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