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The Role of Human LRRK2 in Acute Methylmercury Toxicity in Caenorhabditis elegans

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Abstract

Methylmercury (MeHg) exposure and its harmful effects on the developing brain continue to be a global environmental health concern. Decline in mitochondrial function is central to the toxic effects of MeHg and pathogenesis of mitochondria-related diseases including Parkinson's disease (PD). LRRK2 (Leucine-rich repeat kinase 2) mutation is one of the most common genetic risk factors for PD. In this study, we utilize an acute toxicity model of MeHg exposure in the model organism Caenorhabditis elegans (C. elegans) to compare lifespan, developmental progression, mitochondrial membrane potential and reactive oxygen species (ROS) between the wild-type N2 strain, wild-type LRRK2 transgenic strain (WLZ1), and mutant LRRK2(G2019S) transgenic strain (WLZ3). Additionally, the expression levels of skn-1 and gst-4 were investigated. Our results show that acute MeHg exposure (5 and 10 µM) caused a significant developmental delay in the N2 and WLZ3 worms. Notably, the worms expressing wild-type LRRK2 were resistant to 5 µM MeHg- induced developmental retardation. ROS levels in response to MeHg exposure were increased in the N2 worms, but not in the WLZ1 or WLZ3 worms. The mitochondrial membrane potential was decreased in the N2 worms but increased in the WLZ1 and WLZ3 worms following MeHg exposure. Furthermore, MeHg exposure increased the expression of skn-1 in N2, but not in WLZ1 worms. Although skn-1 expression was increased in the WLZ3 worms following MeHg exposure, gst-4 expression was not induced. Both skn-1 and gst-4 had higher basal expression levels in LRRK2s transgenic than wild-type N2 worms. Knocking down of skn-1 with feeding RNAi had a significant developmental effect in WLZ1 worms; however, the effect was not found in WLZ3 worms. These results suggest that mitochondrial dysfunction and a defect in the SKN-1 signaling in the LRRK2 G2019S worms contribute to the severe developmental delay, establishing a modulatory role of LRRK2 mutation in MeHg-induced acute toxicity.

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References

  1. Sheehan MC, Burke TA, Navas-Acien A, Breysse PN, McGready J, Fox MA (2014) Global methylmercury exposure from seafood consumption and risk of developmental neurotoxicity: a systematic review. Bull World Health Organ 92:254–269f

    Article  PubMed  PubMed Central  Google Scholar 

  2. Aschner M, Clarkson TW (1989) Methyl mercury uptake across bovine brain capillary endothelial cells in vitro: the role of amino acids. Pharmacol Toxicol 64:293–297

    Article  PubMed  CAS  Google Scholar 

  3. Aschner M, Eberle NB, Goderie S, Kimelberg HK (1990) Methylmercury uptake in rat primary astrocyte cultures: the role of the neutral amino acid transport system. Brain Res 521:221–228

    Article  PubMed  CAS  Google Scholar 

  4. Pacyna JM, Sundseth K, Pacyna EG, Jozewicz W, Munthe J, Belhaj M, Aström S (2010) An assessment of costs and benefits associated with mercury emission reductions from major anthropogenic sources. J Air Waste Manag Assoc 60:302–315

    Article  PubMed  CAS  Google Scholar 

  5. Schartup AT, Thackray CP, Qureshi A, Dassuncao C, Gillespie K, Hanke A, Sunderland EM (2019) Climate change and overfishing increase neurotoxicant in marine predators. Nature 572:648–650

    Article  PubMed  CAS  Google Scholar 

  6. Myers GJ, Davidson PW (1998) Prenatal methylmercury exposure and children: neurologic, developmental, and behavioral research. Environ Health Perspect 106(Suppl 3):841–847

    Article  PubMed  PubMed Central  Google Scholar 

  7. Davidson PW, Myers GJ, Weiss B, Shamlaye CF, Cox C (2006) Prenatal methyl mercury exposure from fish consumption and child development: a review of evidence and perspectives from the Seychelles Child Development Study. Neurotoxicology 27:1106–1109

    Article  PubMed  CAS  Google Scholar 

  8. Weiss B, Clarkson TW, Simon W (2002) Silent latency periods in methylmercury poisoning and in neurodegenerative disease. Environ Health Perspect 110(Suppl 5):851–854

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Roos D, Seeger R, Puntel R, Vargas Barbosa N (2012) Role of calcium and mitochondria in MeHg-mediated cytotoxicity. J Biomed Biotechnol 2012:248764

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Nesci S, Trombetti F, Pirini M, Ventrella V, Pagliarani A (2016) Mercury and protein thiols: stimulation of mitochondrial F(1)F(O)-ATPase and inhibition of respiration. Chem Biol Interact 260:42–49

    Article  PubMed  CAS  Google Scholar 

  11. Yin Z, Milatovic D, Aschner JL, Syversen T, Rocha JB, Souza DO, Sidoryk M, Albrecht J, Aschner M (2007) Methylmercury induces oxidative injury, alterations in permeability and glutamine transport in cultured astrocytes. Brain Res 1131:1–10

    Article  PubMed  CAS  Google Scholar 

  12. Ke T, Aschner MJN (2019) Bacteria affect Caenorhabditis elegans responses to MeHg toxicity. Neurotoxicology 75:129–135

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. LeBel CP, Ali SF, McKee M, Bondy SC (1990) Organometal-induced increases in oxygen reactive species: the potential of 2’,7’-dichlorofluorescin diacetate as an index of neurotoxic damage. Toxicol Appl Pharmacol 104:17–24

    Article  PubMed  CAS  Google Scholar 

  14. Mori N, Yasutake A, Marumoto M, Hirayama K (2011) Methylmercury inhibits electron transport chain activity and induces cytochrome c release in cerebellum mitochondria. J Toxicol Sci 36:253–259

    Article  PubMed  CAS  Google Scholar 

  15. Barbosa NV, Nogueira CW, Nogara PA, de Bem AF, Aschner M, Rocha JBT (2017) Organoselenium compounds as mimics of selenoproteins and thiol modifier agents. Metallomics 9:1703–1734

    Article  PubMed  CAS  Google Scholar 

  16. Farina M, Aschner M, Rocha JB (2011) Oxidative stress in MeHg-induced neurotoxicity. Toxicol Appl Pharmacol 256:405–417

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Ke T, Bornhorst J, Schwerdtle T, Santamaría A, Soare FAA, Rocha JB, Farina M, Bowman AB, Aschner MJNR (2020) Therapeutic Efficacy of the N, N′ Bis-(2-Mercaptoethyl) Isophthalamide Chelator for Methylmercury Intoxication in Caenorhabditis elegans.1–12

  18. Caito SW, Zhang Y, Aschner M (2013) Involvement of AAT transporters in methylmercury toxicity in Caenorhabditis elegans. Biochem Biophys Res Commun 435:546–550

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Vanduyn N, Settivari R, Wong G, Nass R (2010) SKN-1/Nrf2 inhibits dopamine neuron degeneration in a Caenorhabditis elegans model of methylmercury toxicity. Toxicol Sci 118:613–624

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Unoki T, Akiyama M, Kumagai Y, Gonçalves FM, Farina M, da Rocha JBT, Aschner M (2018) Molecular pathways associated with methylmercury-induced Nrf2 modulation. Front Genet 9:373

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Ni M, Li X, Yin Z, Jiang H, Sidoryk-Wegrzynowicz M, Milatovic D, Cai J, Aschner M (2010) Methylmercury induces acute oxidative stress, altering Nrf2 protein level in primary microglial cells. Toxicol Sci 116:590–603

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. Hayes JD, Dinkova-Kostova AT (2014) The Nrf2 regulatory network provides an interface between redox and intermediary metabolism. Trends Biochem Sci 39:199–218

    Article  PubMed  CAS  Google Scholar 

  23. Bento-Pereira C, Dinkova-Kostova AT (2020) Activation of transcription factor Nrf2 to counteract mitochondrial dysfunction in Parkinson’s disease. Med Res Rev 41(2):785–802

    Article  PubMed  Google Scholar 

  24. Vorojeikina D, Broberg K, Love TM, Davidson PW, van Wijngaarden E, Rand MD (2017) Editor’s highlight: glutathione S-transferase activity moderates methylmercury toxicity during development in Drosophila. Toxicol Sci 157:211–221

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Ke T, Santamaria A, Rocha JBT, Tinkov AA, Lu R, Bowman AB, Aschner M (2020) The role of human LRRK2 in methylmercury-induced inhibition of microvesicle formation of cephalic neurons in Caenorhabditis elegans. Neurotoxicol Res 38:751–764

    Article  CAS  Google Scholar 

  26. Tolosa E, Vila M, Klein C, Rascol O (2020) LRRK2 in Parkinson disease: challenges of clinical trials. Nat Rev Neurol 16:97–107

    Article  PubMed  Google Scholar 

  27. Howlett EH, Jensen N, Belmonte F, Zafar F, Hu X, Kluss J, Schule B, Kaufman BA, Greenamyre JT, Sanders LH (2017) LRRK2 G2019S-induced mitochondrial DNA damage is LRRK2 kinase dependent and inhibition restores mtDNA integrity in Parkinson’s disease. Hum Mol Genet 26:4340–4351

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Saha S, Guillily MD, Ferree A, Lanceta J, Chan D, Ghosh J, Hsu CH, Segal L, Raghavan K, Matsumoto K, Hisamoto N, Kuwahara T, Iwatsubo T, Moore L, Goldstein L, Cookson M, Wolozin B (2009) LRRK2 modulates vulnerability to mitochondrial dysfunction in Caenorhabditis elegans. J Neurosci 29:9210–9218

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  29. Loeffler DA, Klaver AC, Coffey MP, Aasly JO, LeWitt PA (2017) Increased oxidative stress markers in cerebrospinal fluid from healthy subjects with Parkinson’s disease-associated LRRK2 gene mutations. Front Aging Neurosci 9:89

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  30. Lionaki E, Markaki M, Palikaras K, Tavernarakis N (2015) Mitochondria, autophagy and age-associated neurodegenerative diseases: new insights into a complex interplay. Biochim Biophys Acta 1847:1412–1423

    Article  PubMed  CAS  Google Scholar 

  31. Ke T, Antunes Soares FA, Santamaría A, Bowman AB, Skalny AV, Aschner M (2020) N, N’ bis-(2-mercaptoethyl) isophthalamide induces developmental delay in Caenorhabditis elegans by promoting DAF-16 nuclear localization. Toxicol Rep 7:930–937

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Ke T, Tsatsakis A, Santamaría A, Soare FAA, Tinkov AA, Docea AO, Skalny A, Bowman AB, Aschner MJN (2020) Chronic exposure to methylmercury induces puncta formation in cephalic dopaminergic neurons in Caenorhabditis elegans. Neurotoxicology 7:105–113

    Article  CAS  Google Scholar 

  33. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  34. Shabalina IG, Nedergaard J (2011) Mitochondrial ('mild’) uncoupling and ROS production: physiologically relevant or not? Biochem Soc Trans 39:1305–1309

    Article  PubMed  CAS  Google Scholar 

  35. Hare MF, Atchison WD (1992) Comparative action of methylmercury and divalent inorganic mercury on nerve terminal and intraterminal mitochondrial membrane potentials. J Pharmacol Exp Ther 261:166–172

    PubMed  CAS  Google Scholar 

  36. Papkovskaia TD, Chau KY, Inesta-Vaquera F, Papkovsky DB, Healy DG, Nishio K, Staddon J, Duchen MR, Hardy J, Schapira AH, Cooper JM (2012) G2019S leucine-rich repeat kinase 2 causes uncoupling protein-mediated mitochondrial depolarization. Hum Mol Genet 21:4201–4213

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  37. Mortiboys H, Johansen KK, Aasly JO, Bandmann O (2010) Mitochondrial impairment in patients with Parkinson disease with the G2019S mutation in LRRK2. Neurology 75:2017–2020

    Article  PubMed  CAS  Google Scholar 

  38. Scorrano L, Petronilli V, Bernardi P (1997) On the voltage dependence of the mitochondrial permeability transition pore. A critical appraisal. J Biol Chem 272:12295–12299

    Article  PubMed  CAS  Google Scholar 

  39. Duchen MR (2000) Mitochondria and calcium: from cell signalling to cell death. J Physiol 529(Pt 1):57–68

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. Levesque PC, Atchison WD (1991) Disruption of brain mitochondrial calcium sequestration by methylmercury. J Pharmacol Exp Ther 256:236–242

    PubMed  CAS  Google Scholar 

  41. Padalko VI (2005) Uncoupler of oxidative phosphorylation prolongs the lifespan of Drosophila. Biochemistry (Mosc) 70:986–989

    Article  CAS  Google Scholar 

  42. Marty MS, Atchison WD (1997) Pathways mediating Ca2+ entry in rat cerebellar granule cells following in vitro exposure to methyl mercury. Toxicol Appl Pharmacol 147:319–330

    Article  PubMed  CAS  Google Scholar 

  43. Verma M, Callio J, Otero PA, Sekler I, Wills ZP, Chu CT (2017) Mitochondrial calcium dysregulation contributes to dendrite degeneration mediated by PD/LBD-associated LRRK2 mutants. J Neurosci 37:11151–11165

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  44. Delcambre S, Ghelfi J, Ouzren N, Grandmougin L, Delbrouck C, Seibler P, Wasner K, Aasly JO, Klein C, Trinh J, Pereira SL, Grünewald A (2020) Mitochondrial mechanisms of LRRK2 G2019S penetrance. Front Neurol 11:881

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the National Institutes of Health to MA and ABB (NIEHS R01ES007331). Some strains were provided by the CGC, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440).

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Authors and Affiliations

  1. Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY, 10461, USA

    Tao Ke & Michael Aschner

  2. Department of Biochemistry and Molecular Biology, Federal University of Santa Maria, Santa Maria, RS, Brazil

    Joao B. T. Rocha

  3. IM Sechenov First Moscow State Medical University, Sechenov University, Moscow, Russia

    Alexey A. Tinkov & Michael Aschner

  4. Yaroslavl State University, Yaroslavl, Russia

    Alexey A. Tinkov

  5. K.G. Razumovsky Moscow State University of Technologies and Management, Moscow, Russia

    Alexey A. Tinkov

  6. Laboratorio de Aminoácidos Excitadores, Instituto Nacional de Neurología y Neurocirugía, 14269, Mexico City, Mexico

    Abel Santamaria

  7. School of Health Sciences, Purdue University, West Lafayette, IN, 47907-2051, USA

    Aaron B. Bowman

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  1. Tao Ke
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Correspondence to Tao Ke.

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Ke, T., Rocha, J.B.T., Tinkov, A.A. et al. The Role of Human LRRK2 in Acute Methylmercury Toxicity in Caenorhabditis elegans. Neurochem Res 46, 2991–3002 (2021). https://doi.org/10.1007/s11064-021-03394-y

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  • DOI: https://doi.org/10.1007/s11064-021-03394-y

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