Blocking c-Jun N-terminal Kinase (JNK) Translocation to the Mitochondria Prevents 6-Hydroxydopamine-induced Toxicity in Vitro and in Vivo

• Published in: The Journal of biological chemistry Vol. 288; no. 2; pp. 1079 - 1087
• Main Authors: Chambers, Jeremy W., Howard, Shannon, LoGrasso, Philip V.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 11.01.2013; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Blotting, Western; Cell Death; Cell Line; Dopamine - metabolism; Gene Silencing; Humans; Immunohistochemistry; In Vitro Techniques; JNK Mitogen-Activated Protein Kinases - metabolism; Jun N-terminal Kinase (JNK); Mitochondria - drug effects; Mitochondria - metabolism; Mitochondrial Apoptosis; Neurobiology; Neurons - cytology; Oxidative Stress; Oxidopamine - toxicity; Parkinson Disease; Protein Transport; Rats; Rats, Sprague-Dawley; Signal Transduction; Parkinson Disease; Mitochondrial Apoptosis; Oxidative Stress; Neurobiology; Jun N-terminal Kinase (JNK)
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M112.421354

Because oxidative stress and mitochondrial dysfunction are well known contributors to Parkinson disease (PD), we set out to investigate the role mitochondrial JNK plays in the etiology of 6-hydroxydopamine-induced (6-OHDA) oxidative stress, mitochondrial dysfunction, and neurotoxicity in SHSY5Y cells and neuroprotection and motor behavioral protection in vivo. To do this, we utilized a cell-permeable peptide of the outer mitochondrial membrane protein, Sab (SH3BP5), as an inhibitor of JNK mitochondrial translocation. In vitro studies showed that 6-OHDA induced JNK translocation to the mitochondria and that inhibition of mitochondrial JNK signaling by Tat-SabKIM1 protected against 6-OHDA-induced oxidative stress, mitochondrial dysfunction, and neurotoxicity. Administration of Tat-SabKIM1 via an intracerebral injection into the mid-forebrain bundle increased the number of tyrosine hydroxylase immunoreactive neurons in the substantia nigra pars compacta by 2-fold (p < 0.05) in animals lesioned with 6-OHDA, compared with animals treated only with 6-OHDA into the nigrostriatal pathway. In addition, Tat-SabKIM1 decreased the d-amphetamine-induced unilateral rotations associated with the lesion by 30% (p < 0.05). Steady-state brain levels of Tat-SabKIM1 at day 7 were 750 nm, which was ∼3.4-fold higher than the IC50 for this peptide versus Sab protein. Collectively, these data suggest that 6-OHDA induced JNK translocation to the mitochondria and that blocking this translocation reduced oxidative stress, mitochondrial dysfunction, and neurotoxicity both in vitro and in vivo. Moreover, the data suggest that inhibitors that block association of JNKs with the mitochondria may be useful neuroprotective agents for the treatment of Parkinson disease. Background: Little is known about the role for JNK mitochondrial signaling in neuronal cell death. Results: Global and mitochondrial inhibition of JNK protects against 6-OHDA-induced neuronal loss in the SNpc. Conclusion: Blocking JNK mitochondrial translocation or JNK inhibition may be an effective treatment for neuronal death in Parkinson disease. Significance: These findings suggest a new molecular target for JNK inhibition.