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Inhibition of mixed lineage kinase 3 attenuates MPP+-induced neurotoxicity in SH-SY5Y cells

2004; Elsevier BV; Volume: 1003; Issue: 1-2 Linguagem: Inglês

10.1016/j.brainres.2003.11.073

1872-6240

Joanne R. Mathiasen, Beth Ann McKenna, Michael S. Saporito, Ghanashyam D. Ghadge, Raymond P. Roos, Beverly P. Holskin, Zhiliang Wu, Stephen P. Trusko, Thomas C. Connors, Anna C. Maroney, Beth Ann Thomas, Jeffrey Thomas, Donna Bozyczko‐Coyne,

Signaling Pathways in Disease

The neuropathology of Parkinson's Disease has been modeled in experimental animals following MPTP treatment and in dopaminergic cells in culture treated with the MPTP neurotoxic metabolite, MPP+. MPTP through MPP+ activates the stress-activated c-Jun N-terminal kinase (JNK) pathway in mice and SH-SY5Y neuroblastoma cells. Recently, it was demonstrated that CEP-1347/KT7515 attenuated MPTP-induced nigrostriatal dopaminergic neuron degeneration in mice, as well as MPTP-induced JNK phosphorylation. Presumably, CEP-1347 acts through inhibition of at least one upstream kinase within the mixed lineage kinase (MLK) family since it has been shown to inhibit MLK 1, 2 and 3 in vitro. Activation of the MLK family leads to JNK activation. In this study, the potential role of MLK and the JNK pathway was examined in MPP+-induced cell death of differentiated SH-SY5Y cells using CEP-1347 as a pharmacological probe and dominant negative adenoviral constructs to MLKs. CEP-1347 inhibited MPP+-induced cell death and the morphological features of apoptosis. CEP-1347 also prevented MPP+-induced JNK activation in SH-SY5Y cells. Endogenous MLK 3 expression was demonstrated in SH-SY5Y cells through protein levels and RT-PCR. Adenoviral infection of SH-SY5Y cells with a dominant negative MLK 3 construct attenuated the MPP+-mediated increase in activated JNK levels and inhibited neuronal death following MPP+ addition compared to cultures infected with a control construct. Adenoviral dominant negative constructs of two other MLK family members (MLK 2 and DLK) did not protect against MPP+-induced cell death. These studies show that inhibition of the MLK 3/JNK pathway attenuates MPP+-mediated SH-SY5Y cell death in culture and supports the mechanism of action of CEP-1347 as an MLK family inhibitor.