Abstract
Arsenolipids are lipid-soluble organoarsenic compounds, mainly occurring in marine organisms, with arsenic-containing hydrocarbons (AsHCs) and arsenic-containing fatty acids (AsFAs) representing two major subgroups. Recently, toxicity studies of several arsenolipids showed a high cytotoxic potential of those arsenolipids in human liver and bladder cells. Furthermore, feeding studies with Drosophila melanogaster indicated an accumulation of arsenolipids in the fruit fly’s brain. In this study, the neurotoxic potential of three AsHCs, two AsFAs and three metabolites (dimethylarsinic acid, thio/oxo-dimethylarsenopropanoic acid) was investigated in comparison to the toxic reference arsenite (iAsIII) in fully differentiated human brain cells (LUHMES cells). Thereby, in the case of AsHCs both the cell number and cell viability were reduced in a low micromolar concentration range comparable to iAsIII, while AsFAs and the applied metabolites were less toxic. Mechanistic studies revealed that AsHCs reduced the mitochondrial membrane potential, whereas neither iAsIII nor AsFAs had an impact. Furthermore, neurotoxic mechanisms were investigated by examining the neuronal network. Here, AsHCs massively disturbed the neuronal network and induced apoptotic effects, while iAsIII and AsFAs showed comparatively lesser effects. Taking into account the substantial in vitro neurotoxic potential of the AsHCs and the fact that they could transfer across the physiological barriers of the brain, a neurotoxic potential in vivo for the AsHCs cannot be excluded and needs to be urgently characterized.
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Abbreviations
- AsFA:
-
Arsenic-containing fatty acids
- AsHC:
-
Arsenic-containing hydrocarbons
- AsPL:
-
Arsenosugar-phospholipids
- DMAV :
-
Dimethylarsinic acid
- HepG2:
-
Liver cells
- iAsIII :
-
Arsenite
- LUHMES:
-
Lund human mesencephalic cells
- oxo-DMAPr:
-
Oxo-dimethylarsenopropanoic acid
- thio-DMAPr:
-
Thio-dimethylarsenopropanoic acid
- TMAsFOH:
-
Trimethylarsenio fatty alcohols
- UROtsa:
-
Urothelial cells
References
Amayo KO, Raab A, Krupp EM, Gunnlaugsdottir H, Feldmann J (2013) Novel identification of arsenolipids using chemical derivatizations in conjunction with RP-HPLC-ICPMS/ESMS. Anal Chem 85:9321–9327. doi:10.1021/ac4020935
Amayo KO, Raab A, Krupp EM, Marschall T, Horsfall M Jr, Feldmann J (2014) Arsenolipids show different profiles in muscle tissues of four commercial fish species. J Trace Elem Med Biol 28:131–137. doi:10.1016/j.jtemb.2013.11.004
ATSDR (2016) Addendum to the toxicological profile for arsenic. agency for toxic substances and disease registry (ATSDR), Atlanta, GA
Attene-Ramos MS et al (2013) Systematic study of mitochondrial toxicity of environmental chemicals using quantitative high throughput screening. Chem Res Toxicol 26:1323–1332. doi:10.1021/tx4001754
Aung KH, Kurihara R, Nakashima S, Maekawa F, Nohara K, Kobayashi T, Tsukahara S (2013) Inhibition of neurite outgrowth and alteration of cytoskeletal gene expression by sodium arsenite. Neurotoxicology 34:226–235. doi:10.1016/j.neuro.2012.09.008
Borak J, Hosgood HD (2007) Seafood arsenic: implications for human risk assessment. Regul Toxicol Pharmacol 47:204–212. doi:10.1016/j.yrtph.2006.09.005
Bozyczko-Coyne D, McKenna BW, Connors TJ, Neff NT (1993) A rapid fluorometric assay to measure neuronal survival in vitro. J Neurosci Methods 50:205–216. doi:10.1016/0165-0270(93)90009-G
Brunk CF, Jones KC, James TW (1979) Assay for nanogram quantities of DNA in cellular homogenates. Anal Biochem 92:497–500. doi:10.1016/0003-2697(79)90690-0
Carmeliet P (2003) Blood vessels and nerves: common signals, pathways and diseases. Nat Rev Genet 4:710–720. doi:10.1038/nrg1158
Chattopadhyay S, Bhaumik S, Nag Chaudhury A, Das Gupta S (2002) Arsenic induced changes in growth development and apoptosis in neonatal and adult brain cells in vivo and in tissue culture. Toxicol Lett 128:73–84. doi:10.1016/S0378-4274(01)00535-5
Chazotte B (2011) Labeling mitochondria with MitoTracker dyes. Cold Spring Harb Protoc 2011:990–992. doi:10.1101/pdb.prot5648
Chou CT, Lin HT, Hwang PA, Wang ST, Hsieh CH, Hwang DF (2015) Taurine resumed neuronal differentiation in arsenite-treated N2a cells through reducing oxidative stress, endoplasmic reticulum stress, and mitochondrial dysfunction. Amino Acids 47:735–744. doi:10.1007/s00726-014-1901-1
Cohen SM, Arnold LL, Eldan M, Lewis AS, Beck BD (2006) Methylated arsenicals: the implications of metabolism and carcinogenicity studies in rodents to human risk assessment. Crit Rev Toxicol 36:99–133. doi:10.1080/10408440500534230
Ebert F et al (2014) Toxicological properties of the thiolated inorganic arsenic and arsenosugar metabolite thio-dimethylarsinic acid in human bladder cells. J Trace Elem Med Biol 28:138–146. doi:10.1016/j.jtemb.2013.06.004
Ebert F et al (2016) Evaluating long-term cellular effects of the arsenic species thio-DMA(V): qPCR-based gene expression as screening tool. J Trace Elem Med Biol 37:78–84. doi:10.1016/j.jtemb.2016.06.004
Francesconi AK (2010) Arsenic species in seafood: Origin and human health implications. Pure Appl Chem 82:373–381. doi:10.1351/PAC-CON-09-07-01
Garcia-Salgado S, Quijano MA, Bonilla MM (2012) Arsenic speciation in edible alga samples by microwave-assisted extraction and high performance liquid chromatography coupled to atomic fluorescence spectrometry. Anal Chim Acta 714:38–46. doi:10.1016/j.aca.2011.12.001
Hu D, Kipps TJ (1999) Reduction in mitochondrial membrane potential is an early event in Fas-independent CTL-mediated apoptosis. Cell Immunol 195:43–52. doi:10.1006/cimm.1999.1513
IARC (2012) A review of human carinogens. Part C: arsenic, metals, fibres, and dusts. IARC Monogr 100:196–211
Keil VC, Funke F, Zeug A, Schild D, Muller M (2011) Ratiometric high-resolution imaging of JC-1 fluorescence reveals the subcellular heterogeneity of astrocytic mitochondria. Pflugers Arch 462:693–708. doi:10.1007/s00424-011-1012-8
Korenic A, Boltze J, Deten A, Peters M, Andjus P, Radenovic L (2014) Astrocytic mitochondrial membrane hyperpolarization following extended oxygen and glucose deprivation. PLoS One 9:e90697. doi:10.1371/journal.pone.0090697
Krug AK, Balmer NV, Matt F, Schonenberger F, Merhof D, Leist M (2013) Evaluation of a human neurite growth assay as specific screen for developmental neurotoxicants. Arch Toxicol. doi:10.1007/s00204-013-1072-y
Leffers L, Ebert F, Taleshi MS, Francesconi KA, Schwerdtle T (2013) In vitro toxicological characterization of two arsenosugars and their metabolites. Mol Nutr Food Res 57:1270–1282. doi:10.1002/mnfr.201200821
Lischka S, Arroyo-Abad U, Mattusch J, Kuhn A, Piechotta C (2013) The high diversity of arsenolipids in herring fillet (Clupea harengus). Talanta 110:144–152. doi:10.1016/j.talanta.2013.02.051
Lohren H, Blagojevic L, Fitkau R, Ebert F, Schildknecht S, Leist M, Schwerdtle T (2015) Toxicity of organic and inorganic mercury species in differentiated human neurons and human astrocytes. J Trace Elem Med Biol 32:200–208. doi:10.1016/j.jtemb.2015.06.008
Lotharius J, Barg S, Wiekop P, Lundberg C, Raymon HK, Brundin P (2002) Effect of mutant alpha-synuclein on dopamine homeostasis in a new human mesencephalic cell line. J Biol Chem 277:38884–38894. doi:10.1074/jbc.M205518200
Lotharius J, Falsig J, van Beek J, Payne S, Dringen R, Brundin P, Leist M (2005) Progressive degeneration of human mesencephalic neuron-derived cells triggered by dopamine-dependent oxidative stress is dependent on the mixed-lineage kinase pathway. J Neurosci 25:6329–6342. doi:10.1523/JNEUROSCI.1746-05.2005
Ly JD, Grubb DR, Lawen A (2003) The mitochondrial membrane potential (deltapsi(m)) in apoptosis. An update. Apoptosis 8:115–128. doi:10.1023/A:1022945107762
Maekawa F, Tsuboi T, Oya M, Aung KH, Tsukahara S, Pellerin L, Nohara K (2013) Effects of sodium arsenite on neurite outgrowth and glutamate AMPA receptor expression in mouse cortical neurons. Neurotoxicology 37:197–206. doi:10.1016/j.neuro.2013.05.006
Meyer S, Matissek M, Muller SM, Taleshi MS, Ebert F, Francesconi KA, Schwerdtle T (2014a) In vitro toxicological characterisation of three arsenic-containing hydrocarbons. Metallomics 6:1023–1033. doi:10.1039/c4mt00061g
Meyer S, Schulz J, Jeibmann A, Taleshi MS, Ebert F, Francesconi KA, Schwerdtle T (2014b) Arsenic-containing hydrocarbons are toxic in the in vivo model Drosophila melanogaster. Metallomics 6:2010–2014. doi:10.1039/c4mt00249k
Meyer S et al (2015a) In vitro toxicological characterisation of arsenic-containing fatty acids and three of their metabolites. Toxicol Res (Camb) 4:1289–1296 doi:10.1039/c5tx00122f
Meyer S, Raber G, Ebert F, Taleshi MS, Francesconi KA, Schwerdtle T (2015b) Arsenic-containing hydrocarbons and arsenic-containing fatty acids: Transfer across and presystemic metabolism in the Caco-2 intestinal barrier model. Mol Nutr Food Res 59:2044–2056. doi:10.1002/mnfr.201500286
Namgung U, Xia Z (2001) Arsenic induces apoptosis in rat cerebellar neurons via activation of JNK3 and p38 MAP kinases. Toxicol Appl Pharmacol 174:130–138. doi:10.1006/taap.2001.9200
Niehoff AC et al (2016) Imaging by elemental and molecular mass spectrometry reveals the uptake of an arsenolipid in the brain of Drosophila melanogaster. Anal Chem 88:5258–5263. doi:10.1021/acs.analchem.6b00333
O’Brien J, Wilson I, Orton T, Pognan F (2000) Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 267:5421–5426. doi:10.1046/j.1432-1327.2000.01606.x
Pereira ER et al (2016) Arsenic containing medium and long chain fatty acids in marine fish oil identified as degradation products using reversed-phase HPLC-ICP-MS/ESI-MS. J Anal At Spectrom 31:1836–1845. doi:10.1039/C6JA00162A
Perry SW, Norman JP, Barbieri J, Brown EB, Gelbard HA (2011) Mitochondrial membrane potential probes and the proton gradient: a practical usage guide. Biotechniques 50:98–115. doi:10.2144/000113610
Pöltl D, Schildknecht S, Karreman C, Leist M (2012) Uncoupling of ATP-depletion and cell death in human dopaminergic neurons. Neurotoxicology 33:769–779. doi:10.1016/j.neuro.2011.12.007
Radio NM, Mundy WR (2008) Developmental neurotoxicity testing in vitro: models for assessing chemical effects on neurite outgrowth. Neurotoxicology 29:361–376. doi:10.1016/j.neuro.2008.02.011
Rodriguez VM et al (2005) Glutathione reductase inhibition and methylated arsenic distribution in Cd1 mice brain and liver. Toxicol Sci 84:157–166. doi:10.1093/toxsci/kfi057
Rumpler A et al (2008) Arsenic-containing long-chain fatty acids in cod-liver oil: a result of biosynthetic infidelity? Angew Chem Int Ed Engl 47:2665–2667. doi:10.1002/anie.200705405
Schildknecht S et al (2009) Requirement of a dopaminergic neuronal phenotype for toxicity of low concentrations of 1-methyl-4-phenylpyridinium to human cells. Toxicol Appl Pharmacol 241:23–35. doi:10.1016/j.taap.2009.07.027
Schmeisser E, Goessler W, Francesconi KA (2006) Human metabolism of arsenolipids present in cod liver. Anal Bioanal Chem 385:367–376. doi:10.1007/s00216-006-0401-x
Scholz D, Poltl D, Genewsky A, Weng M, Waldmann T, Schildknecht S, Leist M (2011) Rapid, complete and large-scale generation of post-mitotic neurons from the human LUHMES cell line. J Neurochem 119:957–971. doi:10.1111/j.1471-4159.2011.07255.x
Sele V, Sloth JJ, Lundebye A, Larsen EH, Berntssen MHG, Amlund H (2012) Arsenolipids in marine oils and fats: A review of occurrence, chemistry and future research needs. Food Chem 133:618–630. doi:10.1016/j.foodchem.2012.02.004
Sele V, Sloth JJ, Julshamn K, Skov K, Amlund H (2015) A study of lipid- and water-soluble arsenic species in liver of Northeast Arctic cod (Gadus morhua) containing high levels of total arsenic. J Trace Elem Med Biol 30:171–179. doi:10.1016/j.jtemb.2014.12.010
Su CK, Yang CH, Lin CH, Sun YC (2014) In-vivo evaluation of the permeability of the blood-brain barrier to arsenicals, molybdate, and methylmercury by use of online microdialysis-packed minicolumn-inductively coupled plasma mass spectrometry. Anal Bioanal Chem 406:239–247. doi:10.1007/s00216-013-7429-5
Taleshi MS, Jensen KB, Raber G, Edmonds JS, Gunnlaugsdottir H, Francesconi KA (2008a) Arsenic-containing hydrocarbons: natural compounds in oil from the fish capelin, Mallotus villosus. Chem Commun (Camb). doi:10.1039/b808049f
Taleshi MS, Jensen KB, Raber G, Edmonds JS, Gunnlaugsdottir H, Francesconi KA (2008b) Arsenic-containing hydrocarbons: natural compounds in oil from the fish capelin, Mallotus villosus. Chem Commun (Camb). doi:10.1039/b808049f
Taleshi MS, Raber G, Edmonds JS, Jensen KB, Francesconi KA (2014a) Arsenolipids in oil from blue whiting Micromesistius poutassou—evidence for arsenic-containing esters. Sci Rep 4:1–7 doi:10.1038/srep07492
Taleshi MS, Seidler-Egdal RK, Jensen KB, Schwerdtle T, Francesconi KA (2014b) Synthesis and characterization of arsenolipids: naturally occurring arsenic compounds in fish and algae. Organometallics 33:1397–1403. doi:10.1021/om4011092
Viczek SA, Jensen KB, Francesconi KA (2016) Arsenic-containing phosphatidylcholines: a new group of arsenolipids discovered in herring caviar. Angew Chem Int Ed Engl 55:5259–5262. doi:10.1002/anie.201512031
Wong A, Cortopassi GA (2002) High-throughput measurement of mitochondrial membrane potential in a neural cell line using a fluorescence plate reader. Biochem Biophys Res Commun 298:750–754 pii]
Xi S, Sun W, Wang F, Jin Y, Sun G (2009) Transplacental and early life exposure to inorganic arsenic affected development and behavior in offspring rats. Arch Toxicol 83:549–556. doi:10.1007/s00204-009-0403-5
Acknowledgements
The authors thank Prof. Dr. M. Leist and Dr. Stefan Schildknecht (Doerenkamp-Zbinden Chair of in vitro Toxicology and Biomedicine, University of Konstanz, Germany) for providing the LUHMES cells and supporting the cultivation technique. This work was supported by Deutsche Forschungsgemeinschaft (DFG), Grant Number SCHW903/10-1, the Austrian Science Fund (FWF), Project Number I2412-B21, and the Graduate School of Chemistry (WWU Münster, Germany).
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Witt, B., Meyer, S., Ebert, F. et al. Toxicity of two classes of arsenolipids and their water-soluble metabolites in human differentiated neurons. Arch Toxicol 91, 3121–3134 (2017). https://doi.org/10.1007/s00204-017-1933-x
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DOI: https://doi.org/10.1007/s00204-017-1933-x