title: AMP-activated protein kinase contributes to zinc-induced neuronal death via activation by LKB1 and induction of Bim in mouse cortical cultures.
abstract: BACKGROUND
  We reported that zinc neurotoxicity, a key mechanism of ischemic neuronal death, was mediated by poly ADP-ribose polymerase (PARP) over-activation following NAD(+)/ATP depletion in cortical cultures. Because AMP-activated protein kinase (AMPK) can be activated by ATP depletion, and AMPK plays a key role in excitotoxicity and ischemic neuronal death, we examined whether AMPK could be involved in zinc neurotoxicity in mouse cortical neuronal cultures.
  
  
  RESULTS
  Compound C, an AMPK inhibitor, significantly attenuated zinc-induced neuronal death. Activation of AMPK was detected beginning 2 h after a 10-min exposure of mouse cortical neurons to 300 μM zinc, although a significant change in AMP level was not detected until 4 h after zinc treatment. Thus, AMPK activation might not have been induced by an increase in intracellular AMP in zinc neurotoxicity. Furthermore, we observed that liver kinase B1 (LKB1) but not Ca(2+)/calmodulin-dependent protein kinase kinase β (CaMKKβ), was involved in AMPK activation. Although STO-609, a chemical inhibitor of CaMKKβ, significantly attenuated zinc neurotoxicity, zinc-induced AMPK activation was not affected, which suggested that CaMKKβ was not involved in AMPK activation. Knockdown of LKB1 by siRNA significantly reduced zinc neurotoxicity, as well as zinc-induced AMPK activation, which indicated a possible role for LKB1 as an upstream kinase for AMPK activation. In addition, mRNA and protein levels of Bim, a pro-apoptotic Bcl-2 family member, were noticeably increased by zinc in an AMPK-dependent manner. Finally, caspase-3 activation in zinc-induced neuronal death was mediated by LKB1 and AMPK activation.
  
  
  CONCLUSIONS
  The results suggested that AMPK mediated zinc-induced neuronal death via up-regulation of Bim and activation of caspase-3. Rapid activation of AMPK was detected after exposure of cortical neuronal cultures to zinc, which was induced by LKB1 activation but not increased intracellular AMP levels or CaMKKβ activation. Hence, blockade of AMPK in the brain may protect against zinc neurotoxicity, which is likely to occur after acute brain injury.
authors: Department of Molecular Biology, Sejong University, Seoul, 143-747, South Korea. jweom90@hanmail.net.
authors: Jae-Won
authors: JW
authors: Eom
authors: Department of Molecular Biology, Sejong University, Seoul, 143-747, South Korea. jmineternity80@daum.net.|Department of Integrative Bioscience and Biotechnology, Sejong University, Seoul, 143-747, South Korea. jmineternity80@daum.net.
authors: Jong-Min
authors: JM
authors: Lee
authors: Neural Injury Research Lab & Department of Neurology, University of Ulsan College of Medicine, Seoul, 138-736, South Korea. jkko@amc.seoul.kr.
authors: Jae-Young
authors: JY
authors: Koh
authors: Department of Molecular Biology, Sejong University, Seoul, 143-747, South Korea. yhkim@sejong.ac.kr.|Department of Integrative Bioscience and Biotechnology, Sejong University, Seoul, 143-747, South Korea. yhkim@sejong.ac.kr.
authors: Yang-Hee
authors: YH
authors: Kim
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D000072224:Bcl-2-Like Protein 11; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000473:pathology*; D004789:Enzyme Activation / Q000187:drug effects; D042541:Intracellular Space / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology*; D009498:Neurotoxins / Q000633:toxicity; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D015032:Zinc / Q000633:toxicity*
chemical_list: D051017:Apoptosis Regulatory Proteins; D000072224:Bcl-2-Like Protein 11; C000606776:Bcl2l11 protein, mouse; D008565:Membrane Proteins; D009498:Neurotoxins; D011518:Proto-Oncogene Proteins; D000249:Adenosine Monophosphate; D017346:Protein Serine-Threonine Kinases; C491654:Stk11 protein, mouse; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D055372:AMP-Activated Protein Kinases; D053148:Caspase 3; D015032:Zinc