Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Molecular Brain
Published in: Molecular Brain 1/2016

Open Access 01-12-2016 | Research

AMP-activated protein kinase contributes to zinc-induced neuronal death via activation by LKB1 and induction of Bim in mouse cortical cultures

Authors: Jae-Won Eom, Jong-Min Lee, Jae-Young Koh, Yang-Hee Kim

Published in: Molecular Brain | Issue 1/2016

Login to get access

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.
Literature
1.
Berg JM. Zinc fingers and other metal-binding domains. Elements for interactions between macromolecules. J Biol Chem. 1990;265:6513–6.PubMed
2.
3.
Frederickson CJ, Koh JY, Bush AI. The neurobiology of zinc in health and disease. Nat Rev Neurosci. 2005;6:449–62.CrossRefPubMed
4.
5.
6.
Frederickson CJ, Danscher G. Zinc-containing neurons in hippocampus and related CNS structures. Prog Brain Res. 1990;83:71–84.CrossRefPubMed
7.
Sensi SL, Paoletti P, Bush AI, Sekler I. Zinc in the physiology and pathology of the CNS. Nat Rev Neurosci. 2009;10:780–91.CrossRefPubMed
8.
Qian J, Noebels JL. Exocytosis of vesicular zinc reveals persistent depression of neurotransmitter release during metabotropic glutamate receptor long-term depression at the hippocampal CA3-CA1 synapse. J Neurosci. 2006;26:6089–95.CrossRefPubMed
9.
Kodirov SA, Takizawa S, Joseph J, Kandel ER, Shumyatsky GP, Bolshakov VY. Synaptically released zinc gates long-term potentiation in fear conditioning pathways. Proc Natl Acad Sci U S A. 2006;103:15218–23.PubMedCentralCrossRefPubMed
10.
Koh JY, Suh SW, Gwag BJ, He YY, Hsu CY, Choi DW. The role of zinc in selective neuronal death after transient global cerebral ischemia. Science. 1996;272:1013–6.CrossRefPubMed
11.
Lee SJ, Koh JY. Roles of zinc and metallothionein-3 in oxidative stress-induced lysosomal dysfunction, cell death, and autophagy in neurons and astrocytes. Mol Brain. 2010;3:30.PubMedCentralCrossRefPubMed
12.
13.
Noh KM, Koh JY. Induction and activation by zinc of NADPH oxidase in cultured cortical neurons and astrocytes. J Neurosci. 2000;20:RC111.PubMed
14.
Noh KM, Kim YH, Koh JY. Mediation by membrane protein kinase C of zinc-induced oxidative neuronal injury in mouse cortical cultures. J Neurochem. 1999;72:1609–16.CrossRefPubMed
15.
Seo SR, Chong SA, Lee SI, Sung JY, Ahn YS, Chung KC. Zn2 + −induced ERK activation mediated by reactive oxygen species causes cell death in differentiated PC12 cells. J Neurochem. 2001;78:600–10.CrossRefPubMed
16.
Kim YH, Koh JY. The role of NADPH oxidase and neuronal nitric oxide synthase in zinc-induced poly(ADP-ribose) polymerase activation and cell death in cortical culture. Exp Neurol. 2002;177:407–18.CrossRefPubMed
17.
Kim EY, Koh JY, Kim YH, Sohn S, Joe E, Gwag BJ. Zn2+ entry produces oxidative neuronal necrosis in cortical cell cultures. Eur J Neurosci. 1999;11:327–34.CrossRefPubMed
18.
Kim YH, Kim EY, Gwag BJ, Sohn S, Koh JY. Zinc-induced cortical neuronal death with features of apoptosis and necrosis: mediation by free radicals. Neuroscience. 1999;89:175–82.CrossRefPubMed
19.
Lobner D, Canzoniero LM, Manzerra P, Gottron F, Ying H, Knudson M. Zinc-induced neuronal death in cortical neurons. Cell Mol Biol (Noisy-le-Grand). 2000;46:797–806.
20.
Park JA, Lee JY, Sato TA, Koh JY. Co-induction of p75NTR and p75NTR-associated death executor in neurons after zinc exposure in cortical culture or transient ischemia in the rat. J Neurosci. 2000;20:9096–103.PubMed
21.
Hwang JJ, Lee SJ, Kim TY, Cho JH, Koh JY. Zinc and 4-hydroxy-2-nonenal mediate lysosomal membrane permeabilization induced by H2O2 in cultured hippocampal neurons. J Neurosci. 2008;28:3114–22.CrossRefPubMed
22.
Hardie DG. The AMP-activated protein kinase pathway--new players upstream and downstream. J Cell Sci. 2004;117:5479–87.CrossRefPubMed
23.
Hardie DG, Ross FA, Hawley SA. AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol. 2012;13:251–62.CrossRefPubMed
24.
25.
Carling D, Sanders MJ, Woods A. The regulation of AMP-activated protein kinase by upstream kinases. Int J Obes (Lond). 2008;4(Suppl):S55–59.CrossRef
26.
27.
28.
Sakamoto K, Zarrinpashneh E, Budas GR, Pouleur AC, Dutta A, Prescott AR. Deficiency of LKB1 in heart prevents ischemia-mediated activation of AMPK alpha2 but not AMPK alpha1. Am J Physiol Endocrinol Metab. 2006;290:E780–788.PubMedCentralCrossRefPubMed
29.
Anderson KA, Ribar TJ, Lin F, Noeldner PK, Green MF, Muehlbauer MJ. Hypothalamic CaMKK2 contributes to the regulation of energy balance. Cell Metab. 2008;7:377–88.CrossRefPubMed
30.
Tamas P, Hawley SA, Clarke RG, Mustard KJ, Green K, Hardie DG. Regulation of the energy sensor AMP-activated protein kinase by antigen receptor and Ca2+ in T lymphocytes. J Exp Med. 2006;203:1665–70.PubMedCentralCrossRefPubMed
31.
Stahmann N, Woods A, Carling D, Heller R. Thrombin activates AMP-activated protein kinase in endothelial cells via a pathway involving Ca2+/calmodulin-dependent protein kinase kinase beta. Mol Cell Biol. 2006;26:5933–45.PubMedCentralCrossRefPubMed
32.
Li J, Benashski SE, Siegel C, Liu F, McCullough LD. Adenosine monophosphate activated protein kinase inhibition is protective in both sexes after experimental stroke. Neurosci Lett. 2010;482:62–5.PubMedCentralCrossRefPubMed
33.
Li J, Zeng Z, Viollet B, Ronnett GV, McCullough LD. Neuroprotective effects of adenosine monophosphate-activated protein kinase inhibition and gene deletion in stroke. Stroke. 2007;38:2992–9.PubMedCentralCrossRefPubMed
34.
McCullough LD, Zeng Z, Li H, Landree LE, McFadden J, Ronnett GV. Pharmacological inhibition of AMP-activated protein kinase provides neuroprotection in stroke. J Biol Chem. 2005;280:20493–502.CrossRefPubMed
35.
Chiarugi A. “Simple but not simpler”: toward a unified picture of energy requirements in cell death. FASEB J. 2005;19:1783–8.CrossRefPubMed
36.
Concannon CG, Tuffy LP, Weisova P, Bonner HP, Davila D, Bonner C. AMP kinase-mediated activation of the BH3-only protein Bim couples energy depletion to stress-induced apoptosis. J Cell Biol. 2010;189:83–94.PubMedCentralCrossRefPubMed
37.
Garcia-Gil M, Pesi R, Perna S, Allegrini S, Giannecchini M, Camici M. 5'-aminoimidazole-4-carboxamide riboside induces apoptosis in human neuroblastoma cells. Neuroscience. 2003;117:811–20.CrossRefPubMed
38.
Jung JE, Lee J, Ha J, Kim SS, Cho YH, Bail HH. 5-Aminoimidazole-4-carboxamide-ribonucleoside enhances oxidative stress-induced apoptosis through activation of nuclear factor-kappaB in mouse Neuro 2a neuroblastoma cells. Neurosci Lett. 2004;354:197–200.CrossRefPubMed
39.
Nakatsu Y, Kotake Y, Hino A, Ohta S. Activation of AMP-activated protein kinase by tributyltin induces neuronal cell death. Toxicol Appl Pharmacol. 2008;230:358–63.CrossRefPubMed
40.
Culmsee C, Monnig J, Kemp BE, Mattson MP. AMP-activated protein kinase is highly expressed in neurons in the developing rat brain and promotes neuronal survival following glucose deprivation. J Mol Neurosci. 2001;17:45–58.CrossRefPubMed
41.
42.
Li M, Zhao J, Hu Y, Lu H, Guo J. Oxygen free radicals regulate energy metabolism via AMPK pathway following cerebral ischemia. Neurol Res. 2010;32:779–84.CrossRefPubMed
43.
44.
45.
46.
Shabanzadeh AP, Shuaib A, Yang T, Salam A, Wang CX. Effect of zinc in ischemic brain injury in an embolic model of stroke in rats. Neurosci Lett. 2004;356:69–71.CrossRefPubMed
47.
48.
Weiss JH, Sensi SL, Koh JY. Zn(2+): a novel ionic mediator of neural injury in brain disease. Trends Pharmacol Sci. 2000;21:395–401.CrossRefPubMed
49.
Woods A, Vertommen D, Neumann D, Turk R, Bayliss J, Schlattner U. Identification of phosphorylation sites in AMP-activated protein kinase (AMPK) for upstream AMPK kinases and study of their roles by site-directed mutagenesis. J Biol Chem. 2003;278:28434–42.CrossRefPubMed
50.
51.
Korsse SE, Peppelenbosch MP, van Veelen W. Targeting LKB1 signaling in cancer. Biochim Biophys Acta. 2013;1835:194–210.PubMed
52.
Sapkota GP, Deak M, Kieloch A, Morrice N, Goodarzi AA, Smythe C. Ionizing radiation induces ataxia telangiectasia mutated kinase (ATM)-mediated phosphorylation of LKB1/STK11 at Thr-366. Biochem J. 2002;368:507–16.PubMedCentralCrossRefPubMed
53.
Sapkota GP, Kieloch A, Lizcano JM, Lain S, Arthur JS, Williams MR. Phosphorylation of the protein kinase mutated in Peutz-Jeghers cancer syndrome, LKB1/STK11, at Ser431 by p90(RSK) and cAMP-dependent protein kinase, but not its farnesylation at Cys(433), is essential for LKB1 to suppress cell vrowth. J Biol Chem. 2001;276:19469–82.CrossRefPubMed
54.
Collins SP, Reoma JL, Gamm DM, Uhler MD. LKB1, a novel serine/threonine protein kinase and potential tumour suppressor, is phosphorylated by cAMP-dependent protein kinase (PKA) and prenylated in vivo. Biochem J. 2000;345(Pt 3):673–80.PubMedCentralCrossRefPubMed
55.
Song P, Xie Z, Wu Y, Xu J, Dong Y, Zou MH. Protein kinase Czeta-dependent LKB1 serine 428 phosphorylation increases LKB1 nucleus export and apoptosis in endothelial cells. J Biol Chem. 2008;283:12446–55.PubMedCentralCrossRefPubMed
56.
Hawley SA, Pan DA, Mustard KJ, Ross L, Bain J, Edelman AM. Calmodulin-dependent protein kinase kinase-beta is an alternative upstream kinase for AMP-activated protein kinase. Cell Metab. 2005;2:9–19.CrossRefPubMed
57.
Snider BJ, Gottron FJ, Choi DW. Apoptosis and necrosis in cerebrovascular disease. Ann N Y Acad Sci. 1999;893:243–53.CrossRefPubMed
58.
59.
Blazquez C, Geelen MJ, Velasco G, Guzman M. The AMP-activated protein kinase prevents ceramide synthesis de novo and apoptosis in astrocytes. FEBS Lett. 2001;489:149–53.CrossRefPubMed
60.
Salt IP, Johnson G, Ashcroft SJ, Hardie DG. AMP-activated protein kinase is activated by low glucose in cell lines derived from pancreatic beta cells, and may regulate insulin release. Biochem J. 1998;335(Pt 3):533–9.PubMedCentralCrossRefPubMed
61.
Marsin AS, Bertrand L, Rider MH, Deprez J, Beauloye C, Vincent MF. Phosphorylation and activation of heart PFK-2 by AMPK has a role in the stimulation of glycolysis during ischaemia. Curr Biol. 2000;10:1247–55.CrossRefPubMed
62.
Kleman AM, Yuan JY, Aja S, Ronnett GV, Landree LE. Physiological glucose is critical for optimized neuronal viability and AMPK responsiveness in vitro. J Neurosci Methods. 2008;167:292–301.PubMedCentralCrossRefPubMed
63.
Zhu J, Aja S, Kim EK, Park MJ, Ramamurthy S, Jia J. Physiological oxygen level is critical for modeling neuronal metabolism in vitro. J Neurosci Res. 2012;90:422–34.CrossRefPubMed
64.
65.
Emerling BM, Weinberg F, Snyder C, Burgess Z, Mutlu GM, Viollet B. Hypoxic activation of AMPK is dependent on mitochondrial ROS but independent of an increase in AMP/ATP ratio. Free Radic Biol Med. 2009;46:1386–91.PubMedCentralCrossRefPubMed
66.
Park JA, Koh JY. Induction of an immediate early gene egr-1 by zinc through extracellular signal-regulated kinase activation in cortical culture: its role in zinc-induced neuronal death. J Neurochem. 1999;73:450–6.CrossRefPubMed
67.
Zhu H, Moriasi CM, Zhang M, Zhao Y, Zou MH. Phosphorylation of serine 399 in LKB1 protein short form by protein kinase Czeta is required for its nucleocytoplasmic transport and consequent AMP-activated protein kinase (AMPK) activation. J Biol Chem. 2013;288:16495–505.PubMedCentralCrossRefPubMed
68.
Lutzner N, Kalbacher H, Krones-Herzig A, Rosl F. FOXO3 is a glucocorticoid receptor target and regulates LKB1 and its own expression based on cellular AMP levels via a positive autoregulatory loop. PLoS One. 2012;7:e42166.PubMedCentralCrossRefPubMed
69.
Davila D, Connolly NM, Bonner H, Weisova P, Dussmann H, Concannon CG. Two-step activation of FOXO3 by AMPK generates a coherent feed-forward loop determining excitotoxic cell fate. Cell Death Differ. 2012;19:1677–88.PubMedCentralCrossRefPubMed
70.
Kilbride SM, Farrelly AM, Bonner C, Ward MW, Nyhan KC, Concannon CG. AMP-activated protein kinase mediates apoptosis in response to bioenergetic stress through activation of the pro-apoptotic Bcl-2 homology domain-3-only protein BMF. J Biol Chem. 2010;285:36199–206.PubMedCentralCrossRefPubMed
71.
Koh JY, Choi DW. Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Methods. 1987;20:83–90.CrossRefPubMed
72.
Fei F, Bowdish DM, McCarry BE. Comprehensive and simultaneous coverage of lipid and polar metabolites for endogenous cellular metabolomics using HILIC-TOF-MS. Anal Bioanal Chem. 2014;406:3723–33.PubMedCentralCrossRefPubMed
73.
Park H, Eom JW, Kim YH. Consensus scoring approach to identify the inhibitors of AMP-activated protein kinase alpha2 with virtual screening. J Chem Inf Model. 2014;54:2139–46.CrossRefPubMed
Metadata
Title
AMP-activated protein kinase contributes to zinc-induced neuronal death via activation by LKB1 and induction of Bim in mouse cortical cultures
Authors
Jae-Won Eom
Jong-Min Lee
Jae-Young Koh
Yang-Hee Kim
Publication date
01-12-2016
Publisher
BioMed Central
Published in
Molecular Brain / Issue 1/2016
Electronic ISSN: 1756-6606
DOI
https://doi.org/10.1186/s13041-016-0194-6

Other articles of this Issue 1/2016

Molecular Brain 1/2016 Go to the issue

Research

Wnt3a upregulates brain-derived insulin by increasing NeuroD1 via Wnt/β-catenin signaling in the hypothalamus

Research

Orchestrated activation of mGluR5 and CB1 promotes neuroprotection

Research

Disrupted-in-schizophrenia 1 (DISC1) and Syntaphilin collaborate to modulate axonal mitochondrial anchoring

Research

Increased phosphorylation of collapsin response mediator protein-2 at Thr514 correlates with β-amyloid burden and synaptic deficits in Lewy body dementias

Research

Personalized medicine approach confirms a milder case of ABAT deficiency

Research

Rescuing macrophage normal function in spinal cord injury with embryonic stem cell conditioned media