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Translocation and oligomerization of Bax is regulated independently by activation of p38 MAPK and caspase-2 during MN9D dopaminergic neurodegeneration

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Abstract

Bax is translocated into the mitochondrial membrane and oligomerized therein to initiate mitochondrial apoptotic signaling. Our previous study indicated that reactive oxygen species (ROS)-mediated activation of mitogen-activated protein kinase (MAPK) and caspase is critically involved in 6-hydroxydopamine (6-OHDA)-mediated neurodegeneration. Here, we specifically attempted to examine whether and how these death signaling pathways may be linked to Bax translocation and oligomerization. We found that 6-OHDA treatment triggered translocation and oligomerization of Bax onto the mitochondria in MN9D dopaminergic neuronal cells. These events preceded cytochrome c release into the cytosol. Cross-linking assay revealed that co-treatment with a ROS scavenger or a pan-caspase inhibitor inhibited 6-OHDA-induced Bax oligomerization. Among several candidates of ROS-activated MAPKs and caspases, we found that co-treatment with PD169316 or VDVAD specifically inhibited 6-OHDA-induced Bax oligomerization, suggesting critical involvement of p38 MAPK and caspase-2. Consequently, overexpression of a dominant negative form of p38 MAPK or a shRNA-mediated knockdown of caspase-2 indeed inhibited 6-OHDA-induced Bax oligomerization. However, activation of p38 MAPK and caspase-2 was independently linked to oligomerization of Bax. This specificity was largely confirmed with a Bax 6A7 antibody known to detect activated forms of Bax on the mitochondria. Taken together, our data suggest that there is an independent amplification loop of Bax translocation and oligomerization via caspase-2 and p38 MAPK during ROS-mediated dopaminergic neurodegeneration.

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Acknowledgments

This work was supported by a grant of the Korea Healthcare Technology R&D Project from the Ministry for Health, Welfare & Family Affairs (A090063), and in part by a grant from the Ministry of Science and Technology through the Brain Research Center (Infra-2), KOSEF (SRC, R11-20080036-00000-0) and WCU (R33-2008-10014).

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Authors and Affiliations

  1. Department of Biology, Yonsei University College of Life Science and Biotechnology, 134 Shinchon-dong Seodaemoon-gu, Seoul, 120-749, Korea

    Chang-Ki Oh, Won-Seok Choi & Young J. Oh

  2. Brain Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, Korea

    Baek-Soo Han

  3. Israel Technion Institute of Technology, Haifa, Israel

    Moussa B. H. Youdim

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  1. Chang-Ki Oh
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Correspondence to Young J. Oh.

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Chang-Ki Oh and Baek-Soo Han contributed equally to this work.

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Oh, CK., Han, BS., Choi, WS. et al. Translocation and oligomerization of Bax is regulated independently by activation of p38 MAPK and caspase-2 during MN9D dopaminergic neurodegeneration. Apoptosis 16, 1087–1100 (2011). https://doi.org/10.1007/s10495-011-0627-8

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