Skip to main content

Advertisement

Springer Nature Link
Log in

Protein profiling as early detection biomarkers for TiO2 nanoparticle toxicity in Daphnia magna

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

The mode of action for nanoparticle (NP) toxicity in aquatic organisms is not yet fully understood. In this work, a strategy other than toxicity testing was applied to Daphnia magna exposed to TiO2-NPs: the use of nuclear microscopy and the assessment of protein profile. D. magna is a keystone species broadly used as a model system in ecotoxicology. Titanium (Ti) was found in the D. magna digestive tract, mainly in the gut. The penetration of Ti into the epithelial region was greater at higher exposure levels and also observed in eggs in the brood pouch. The protein profile of individuals exposed to different concentrations showed that 2.8 and 5.6 mg/L TiO2-NP concentrations induced an over-expression of the majority of proteins, in particular proteins with molecular weight of ∼120, 85 and 15 kDa, while 11.2 mg/L TiO2-NP had an inhibitory effect on protein expression. The Matrix-assisted laser desorption ionization with tandem time of flight mass spectrometry (MALDI-TOF/TOF MS) analysis of these proteins consistently identified them as vitellogenin (Vtg)-like proteins, associated with enzymes involved in redox balance. These results indicate that Vtg-like proteins are up-regulated in D. magna exposed to TiO2-NPs. Vitellogenesis is associated with the reproduction system, suggesting that TiO2-NP exposure can impair reproduction by affecting this process. The precise mode of action of TiO2-NPs is still unclear and the results from this study are a first attempt to identify specific proteins as potential markers of TiO2-NP toxicity in D. magna, providing useful information for future research.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Aguer P, Alves LC, Ph Barberet, Gontier E, Incerti S, Michelet-Habchi C, Kertész Zs, Kiss AZ, Moretto P, Pallon J, Pinheiro T, Surlève-Bazeille JE, Szikszai Z, Veríssimo A, Ynsa MD (2005) Skin morphology and layer identification using different STIM geometries. Nucl Instrum Methods Phys Res B 231:292–299

    Article  CAS  Google Scholar 

  • Aruoja V, Dubourguier H-C, Kasemets K, Kahru A (2009) Toxicity of nanoparticles of CuO, ZnO and TiO2 to microalgae Pseudokirchneriella subcapitata. Sci Total Environ 407:1461–1468

    Article  CAS  Google Scholar 

  • Bradford MM (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Bradley BP, Kalampanayil B, O’Neill MC (2009) Protein expression profiling. Methods Mol Biol 519:455–468

    Article  CAS  Google Scholar 

  • Breese MB, Jamieson DN, King PJ (1996) Materials analysis using a nuclear microprobe. John Wiley & Sons Ltd, Chichester, Sussex, Journals, Bans Lane PO 19 1 UD, UK, 1996. 428

    Google Scholar 

  • Chen S, Chen DF, Yang F, Nagasawa H, Yang WJ (2011) Characterization and processing of superoxide dsmutase-fused vitellogenin in the diapause embryo formation: a special developmental pathway in the brine shrimp, Artemia parthenogenetica. Biol Reprod 85:31–41

    Article  CAS  Google Scholar 

  • Chevalier J, Harscoët E, Keller M, Pandard P, Cachot J, Grote M (2015) Exploration of Daphnia behavioral effect profiles induced by a broad range of toxicants with different modes of action. Environ Toxicol Chem 34:1760–1769

    Article  CAS  Google Scholar 

  • Colbourne JK, Pfrender ME, Gilbert D, Thomas WK, Tucker A, Oakley TH et al. (2011) The Ecoresponsive genome of Daphnia pulex. Science 331:555–561

    Article  CAS  Google Scholar 

  • Dabrunz A, Duester L, Prasse C, Seitz F, Rosenfeldt R, Schilde C, Schaumann GE, Schulz R (2011) Biological surface coating and molting inhibition as mechanisms of TiO2 nanoparticle toxicity in Daphnia magna. PLoS ONE 6:e20112

    Article  CAS  Google Scholar 

  • de Chaffoy CD, Kondo M (1980) Lipovitellin from the crustacean, Artemia salina: biochemical analysis of lipovitellin complex from the yolk granules. J Biol Chem 255:6727–6733

    Google Scholar 

  • Dhadialla TS, Raikhel AS (1990) Biosynthesis of mosquito vitellogenin. J Biol Chem 265:9924–9933

    CAS  Google Scholar 

  • Diniz MS, Matos AP, Lourenço J, Castro L, Peres I, Mendonça E, Picado A (2013) Liver alterations in two freshwater fish species (Carassius auratus and Danio rerio) following exposure to different TiO2 nanoparticles concentrations. Microsc Microanal 19:1131–1140

    Article  CAS  Google Scholar 

  • Elendt BP, Bias WR (1990) Trace nutrient deficiency in Daphnia magna cultured in standard medium for toxicity testing. Effects of the optimization of culture conditions of life history parameters of Daphnia magna. Water Res 24:1157–1167

    Article  CAS  Google Scholar 

  • Exbrayat JM, Moudilou EN, Lapied E (2015) Harmful effects of nanoparticles on animals. J Nanotechnol 2015:1–10

    Article  Google Scholar 

  • Farhadian S, Shareghi B, Salavati-Niasari M, Amooaghaei R (2012) Spectroscopic studies on the interaction of nano-TiO2 with lysozyme. J Nanostr 1:95–103

    Google Scholar 

  • Federici G, Shaw BJ, Handy R (2007) Toxicity of titanium dioxide nanoparticles to rainbow trout (Oncorhynchus mykiss): gill injury, oxidative stress, and other physiological effects. Aquat Toxicol 84:415–430

    Article  CAS  Google Scholar 

  • Galesio M, Vieira DV, Rial-Otero R, Lodeiro C, Moura I, Capelo JL (2008) Influence of the protein staining in the fast ultrasonic sample treatment for protein identification through peptide mass fingerprint and matrix-assisted laser desorption ionization time of flight mass spectrometry. J Proteome Res 7:2097–2106

    Article  CAS  Google Scholar 

  • Gopalan R, Osman I, Amani A, de Matas M, Anderson D (2009) The effect of zinc oxide and titanium dioxide nanoparticles in the comet assay with UVA photoactivation of human sperm and lymphocytes. Nanotoxicology 3:33–39

    Article  CAS  Google Scholar 

  • Grime GW, Dawson M (1995) Recent developments in data acquisition and processing on the Oxford scanning proton microprobe. Nucl Instrum Methods Phys Res B 104:107–113

    Article  CAS  Google Scholar 

  • Grime GW (1996) The “Q factor” method: quantitative microPIXE analysis using RBS normalisation. Nucl Instrum Methods Phys Res B 109–110:170–174

    Article  Google Scholar 

  • Guilhermino L, Diamantino T, Silva MS, Soares AMVM (2000) Acute toxicity test with Daphnia magna: an alternative to mammals in the prescreening of chemical toxicity? Ecotoxicol Environ Saf 46:357–362

    Article  CAS  Google Scholar 

  • Gulson B, McCall M, Korsch M, Gomez L, Casey P, Oytam Y, Taylor A, McCulloch M, Trotter J, Kinsley L, Greenoak G (2010) Small amounts of zinc from zinc oxide particles in sunscreens applied outdoors are absorbed through human skin. Toxicol Sci 118:140–149

    Article  CAS  Google Scholar 

  • Gulson B, McCall MJ, Bowman DM, Pinheiro T (2015) A review of critical factors for assessing the dermal absorption of metal oxide nanoparticles from sunscreens applied to humans, and a research strategy to address current deficiencies. Arch Toxicol 89:1909–1930

    Article  CAS  Google Scholar 

  • Havukainen H, Halskau Y, Skjaerven L, Smedal B, Amdam GV (2011) Deconstructing honeybee vitellogenin: novel 40 kDa fragment assigned to its N terminus. J Exp Biol 214:582–592

    Article  CAS  Google Scholar 

  • Hund-Rinke K, Simon M (2006) Ecotoxic effect of photocatalytic active nanoparticles (TiO2) on algae and daphnids. Environ Sci Pollut Res 13:225–232

    Article  CAS  Google Scholar 

  • ISO 6341 (1996) Water quality – determination of the inhibition of the mobility of Daphnia magna straus (Cladocera, Crustacea) – Acute toxicity Test. International Organization for Standardization, ISO 6341

  • Ivask A, Juganson K, Bondarenko O, Mortimer M, Aruoja V, Kasemets K, Blinova I, Heinlaan M, Slaveykova V, Kahru A (2014) Mechanisms of toxic action of Ag, ZnO and CuO nanoparticles to selected ecotoxicological test organisms and mammalian cells in vitro: a comparative review. Nanotoxicology Suppl 1:57–71

    Article  Google Scholar 

  • Kahru A, Dubourguier HC (2010) From ecotoxicology to nanoecotoxicology. Toxicology 269:105–119

    Article  CAS  Google Scholar 

  • Kang HY (2010) A review of the emerging nanotechnology industry: materials, fabrications, and applications. Department of Toxic Substances Control Pollution Prevention and Green Technology, California, p 64

  • Kato Y, Tokishita S, Ohta T, Yamagata H (2004) A vitellogenin chain containing a superoxide dismutase-like domain is the major component of yolk proteins in cladoceran crustacean Daphnia magna. Gene 334:157–165

    Article  CAS  Google Scholar 

  • Kemp CJ, Kültz D (2012) Controlling proteome degradation in Daphnia pulex. J Exp Zool Part A 317A:645–651

    Google Scholar 

  • Khanna P, Ong C, Bay BH, Baeg GH (2015) Nanotoxicity: an interplay of oxidative stress, inflammation and cell death. Nanomaterials 5:1163–1180

    Article  CAS  Google Scholar 

  • Kruger N (1994) The Bradford method for protein quantitation. In: Walker, J (ed) Basic Prot Pep Prot Humana Press, Department of Plant Sciences University of Oxford, UK, pp 9–15

  • Lavicoli I, Fontana L, Leso V, Bergamaschi A (2013) The effects of nanomaterials as endocrine disruptors. Int J Mol Sci 14:16732–16801

    Article  Google Scholar 

  • Li B, Ze YG, Sun QQ, Zhang T, Sang XZ, Cui YL, Wang XC, Gui SX, Tan DL, Zhu M, Zhao XY, Sheng L, Wang L, Hong FS, Tang M (2013) Molecular mechanisms of nanosized titanium dioxide-induced pulmonary injury in mice. PloS ONE 8(2):e55563

    Article  CAS  Google Scholar 

  • Lovern SB, Klaper R (2006) Daphnia magna mortality when exposed to titanium dioxide and fullerene (C-60) nanoparticles. Environ Toxicol Chem 25:1132–1137

    Article  CAS  Google Scholar 

  • Menard A, Drobne D, Jamec A (2011) Ecotoxicity of nanosized TiO2. Review of in vitro data. Env Poll 159:677–684

    Article  CAS  Google Scholar 

  • Meusy JJ, Payen GG (1988) Female reproduction in malacostracan Crustacea. Zool Sci 5:217–265

    Google Scholar 

  • Ni M, Li F, Wang B, Xu K, Zhang H, Hu J, Tian J, Shen W, Li B (2015) Effect of TiO2 nanoparticles on the reproduction of silkworm. Biol Trace Elem Res 164:106–113

    Article  CAS  Google Scholar 

  • Peters R, De Bernardi R (eds) (1987) Daphnia. Memorie Dell’Istituto Italiano di Idrobiologia Dott. Marco de Marchi, 45, p 502, Consiglio Nazionale delle Ricerche, Verbania Pallanza

  • Pinheiro T, Moita L, Silva L, Mendonça E, Picado A (2013) Nuclear microscopy as a tool in TiO2 nanoparticles bioaccumulation studies in aquatic species. Nucl Instrum Methods Phys Res B 306:117–120

    Article  CAS  Google Scholar 

  • Pinheiro T, Silva R, Fleming R, Gonçalves A, Barreiros MA, Silva JN, Morlière P, Santus R, Filipe P (2014) Primary hemochromatosis: distribution and quantitation of skin iron. Correlations with total body stores. Acta Derm-Venereol 94:14–19

    Article  Google Scholar 

  • Raviv S, Parnes S, Segall C, Davis C, Sagi A (2006) Complete sequence of Litopenaeus vannamei (Crustacea: Decapoda) vitellogenin cDNA and its expression in endocrinologically induced sub-adult females. Gen Comp Endocrin 145:39–50

    Article  CAS  Google Scholar 

  • Reith M, Munholland J, Kelly J, Finn RN, Fyhn HJ (2001) Lipovitellins derived from two forms of vitellogenin are differentially processed during oocyte maturation in haddock (Melanogrammus aeglefinus). J Exp Zool 291:58–67

    Article  CAS  Google Scholar 

  • Sappington TW, Raikhel AS (1998) Molecular characteristics of insect vitellogenins and vitellogenin receptors. Insect Biochem Mol Biol 28:277–300

    Article  CAS  Google Scholar 

  • Schlich K, Terytze K, Hund-Rinke K (2012) Effect of TiO2 nanoparticles in the earthworm reproduction test. Environ Sci Eur Bridg Sci Regul Reg Eur Lev 24:5

    Google Scholar 

  • Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M (2006) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Prot 1:2856–2860

    Article  CAS  Google Scholar 

  • Shi H, Magaye R, Castranova V, Zha J (2013) Titanium dioxide nanoparticles: a review of current toxicological data. Part Fibre Toxicol 10:15

    Article  CAS  Google Scholar 

  • Stein JR (1973) Handbook of Phycological Methods, Culture Methods, and Growth Measurements. Cambridge Univ. Press, London

    Google Scholar 

  • Takeda K, Suzuki KI, Ishihara A, Kubo-Irie M, Fujimoto R, Tabata M, Oshio S, Nihei Y, Ihara T, Sugamata M (2009) Nanoparticles transferred from pregnant mice to their offspring can damage the genital and cranial nerve systems. J Health Sci 55:95–102

    Article  CAS  Google Scholar 

  • Tokishita S, Kato Y, Kobayashi T, Nakamura S, Ohta T, Yamagata H (2006) Organization and repression by juvenile hormone of a vitellogenin gene cluster in the crustacean, Daphnia magna. Biochem Biophys Res Commun 345:362–370

    Article  CAS  Google Scholar 

  • Vasco MS, Alves LC, Corregidor V, Bettiol A, Watt F, Pinheiro T (2017) 3D map distribution of metallic nanoparticles in whole cells using MeV ion microscopy. J Microsc (doi: 10.1111/jmi.12561)

  • Veríssimo A, Alves LC, Filipe P, Silva JN, Silva R, Ynsa MD, Gontier E, Moretto P, Pallon J, Pinheiro T (2007) Nuclear Microscopy: a tool for imaging elemental distribution and percutaneous absorption in vivo. Microsc Res Tech 70:302–309

    Article  Google Scholar 

  • Wang J, Zhu X, Zhang X, Zhao Z, Liu H, George R, Wilson-Rawls J, Chang Y, Chen Y (2011) Disruption of zebrafish (Danio rerio) reproduction upon chronic exposure to TiO2 nanoparticles Chemosphere 83:461–467. https://doi.org/10.1016/j.chemosphere 2010.12.069

    Article  CAS  Google Scholar 

  • Wiench K, Wohlleben W, Hisgen V, Radke K, Salinas E, Zok S, Landsiedel R (2009) Acute and chronic effects of nano- and non-nano-scale TiO2 and ZnO particles on mobility and reproduction of the freshwater invertebrate Daphnia magna. Chemosphere 76:1356–1365

    Article  CAS  Google Scholar 

  • Zhang H, Ji Z, Xia T, Meng H, Low-Kam C, Liu R, Pokhrel S, Lin S, Wang X, Liao YP, Wang M, Li L, Rallo R, Damoiseaux R, Telesca D, Mädler L, Cohen Y, Zink JI, Nel AE (2012) Use of metal oxide nanoparticle band gap to develop a predictive paradigm for oxidative stress and acute pulmonary inflammation. ACS Nano 22 6(5):4349–4368

    Article  CAS  Google Scholar 

  • Zhu X, Chang Y, Chen Y (2010) Toxicity and bioaccumulation of TiO2 nanoparticle aggregates in Daphnia magna. Chemosphere 78:209–215

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Project funded by Fundação para a Ciência e Tecnologia, PTDC/CTM/099446/2008 and PEst-C/EQB/LA0006/2011 granted to Requimte. This work was also financed by national funds through FCT–Foundation for Science and Technology, under the project UID/BIO/04565/2013 and Programa Operacional Regional de Lisboa (LISBOA-01-0145-FEDER-007317). In addition, the authors would like to acknowledge Dr. José Carlos Roseiro, senior researcher from LNEG, and Dr. Andrew Wilbey, professor from University of Reading at UK, for carefully proof reading of the English in this article.

Author information

Authors and Affiliations

  1. INIAV—Instituto Nacional de Investigação Agrária e Veterinária, IP, Av. da República, Quinta do Marquês, 2784-505, Oeiras, Portugal

    Paula Sá-Pereira

  2. REQUIMTE, Departamento de Química, Centro de Química Fina e Biotecnologia, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516, Caparica, Portugal

    Mário S. Diniz

  3. LNEG–Laboratório Nacional de Energia e Geologia, IP, Estrada da Portela, Bairro do Zambujal Ap 7586, 2720-999, Amadora, Portugal

    Liliana Moita, Elsa Mendonça, Susana M. Paixão & Ana Picado

  4. Instituto de Bioengenharia e Biociências, Departamento de Engenharia e Ciências Nucleares, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais 1, 1049-001, Lisboa, Portugal

    Teresa Pinheiro

  5. APA-Agência Portuguesa do Ambiente, I.P. Rua da Murgueira 9/9ª, 2610-124, Amadora, Portugal

    Elsa Mendonça

Authors
  1. Paula Sá-Pereira
    View author publications

    Search author on:PubMed Google Scholar

  2. Mário S. Diniz
    View author publications

    Search author on:PubMed Google Scholar

  3. Liliana Moita
    View author publications

    Search author on:PubMed Google Scholar

  4. Teresa Pinheiro
    View author publications

    Search author on:PubMed Google Scholar

  5. Elsa Mendonça
    View author publications

    Search author on:PubMed Google Scholar

  6. Susana M. Paixão
    View author publications

    Search author on:PubMed Google Scholar

  7. Ana Picado
    View author publications

    Search author on:PubMed Google Scholar

Corresponding authors

Correspondence to Paula Sá-Pereira or Ana Picado.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sá-Pereira, P., Diniz, M.S., Moita, L. et al. Protein profiling as early detection biomarkers for TiO2 nanoparticle toxicity in Daphnia magna. Ecotoxicology 27, 430–439 (2018). https://doi.org/10.1007/s10646-018-1907-7

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-018-1907-7

Keywords