1. AMP-activated protein kinase contributes to zinc-induced neuronal death via activation by LKB1 and induction of Bim in mouse cortical cultures
- Author
-
Jae Won Eom, Jae-Young Koh, Yang-Hee Kim, and Jong Min Lee
- Subjects
0301 basic medicine ,Adenosine monophosphate ,AMPK ,LKB1 ,Poly ADP ribose polymerase ,Neurotoxins ,Neuronal death ,Excitotoxicity ,Intracellular Space ,Calcium-Calmodulin-Dependent Protein Kinase Kinase ,Brain ischemia ,Bim ,Caspase-3 ,Zinc ,AMP-Activated Protein Kinases ,Protein Serine-Threonine Kinases ,medicine.disease_cause ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Mice ,AMP-activated protein kinase ,Proto-Oncogene Proteins ,medicine ,Animals ,Protein kinase A ,Molecular Biology ,Cells, Cultured ,Cerebral Cortex ,Neurons ,biology ,Bcl-2-Like Protein 11 ,Cell Death ,Kinase ,Caspase 3 ,Research ,Neurotoxicity ,Membrane Proteins ,medicine.disease ,Adenosine Monophosphate ,Cell biology ,Enzyme Activation ,030104 developmental biology ,chemistry ,biology.protein ,Apoptosis Regulatory Proteins - 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 Ca2+/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.
- Published
- 2015