Amino-modified polystyrene nanoparticles affect signalling pathways of the sea urchin (Paracentrotus lividus) embryos

Abstract Polystyrene nanoparticles have been shown to pose serious risk to marine organisms including sea urchin embryos based on their surface properties and consequently behaviour in natural sea water. The aim of this study is to investigate the toxicity pathways of amino polystyrene nanoparticles (PS-NH2, 50 nm) in Paracentrotus lividus embryos in terms of development and signalling at both protein and gene levels. Two sub-lethal concentrations of 3 and 4 μg/mL of PS-NH2 were used to expose sea urchin embryos in natural sea water (PS-NH2 as aggregates of 143 ± 5 nm). At 24 and 48 h post-fertilisation (hpf) embryonic development was monitored and variations in the levels of key proteins involved in stress response and development (Hsp70, Hsp60, MnSOD, Phospho-p38 Mapk) as well as the modulation of target genes (Pl-Hsp70, Pl-Hsp60, Pl-Cytochrome b, Pl-p38 Mapk, Pl-Caspase 8, Pl-Univin) were measured. At 48 hpf various striking teratogenic effects were observed such as the occurrence of cells/masses randomly distributed, severe skeletal defects and delayed development. At 24 hpf a significant up-regulation of Pl-Hsp70, Pl-p38 Mapk, Pl-Univin and Pl-Cas8 genes was found, while at 48 hpf only for Pl-Univin was observed. Protein profile showed different patterns as a significant increase of Hsp70 and Hsp60 only after 48 hpf compared to controls. Conversely, P-p38 Mapk protein significantly increased at 24 hpf and decreased at 48 hpf. Our findings highlight that PS-NH2 are able to disrupt sea urchin embryos development by modulating protein and gene profile providing new understandings into the signalling pathways involved.

[1]  D. Minetto,et al.  Ecotoxicity of engineered TiO2 nanoparticles to saltwater organisms: an overview. , 2014, Environment international.

[2]  Ellen Besseling,et al.  Nanoplastics in the Aquatic Environment. Critical Review , 2015 .

[3]  Andrea Bodnar,et al.  Lessons From the Sea: Marine Animals Provide Models for Biomedical Research , 2016 .

[4]  Bernd Bukau,et al.  The Hsp70 and Hsp60 Chaperone Machines , 1998, Cell.

[5]  T. Snell,et al.  Assessing toxicity of nanoparticles using Brachionus manjavacas (Rotifera) , 2011, Environmental toxicology.

[6]  Ilaria Corsi,et al.  Effects of nanomaterials on marine invertebrates. , 2016, The Science of the total environment.

[7]  C. Bradham,et al.  Late Alk4/5/7 signaling is required for anterior skeletal patterning in sea urchin embryos , 2015, Development.

[8]  A. Pinsino,et al.  Rapid changes in heat-shock cognate 70 levels, heat-shock cognate phosphorylation state, heat-shock transcription factor, and metal transcription factor activity levels in response to heavy metal exposure during sea urchin embryonic development , 2011, Ecotoxicology.

[9]  Franz Brümmer,et al.  Silver nanoparticle toxicity in sea urchin Paracentrotus lividus. , 2013, Environmental pollution.

[10]  K. Dawson,et al.  Evidence for immunomodulation and apoptotic processes induced by cationic polystyrene nanoparticles in the hemocytes of the marine bivalve Mytilus. , 2015, Marine environmental research.

[11]  A A Koelmans,et al.  Effects of nanopolystyrene on the feeding behavior of the blue mussel (Mytilus edulis L.) , 2012, Environmental toxicology and chemistry.

[12]  Andrew R. Jackson,et al.  The Genome of the Sea Urchin Strongylocentrotus purpuratus , 2006, Science.

[13]  H. Zhu,et al.  Identification of a cytochrome b-type NAD(P)H oxidoreductase ubiquitously expressed in human cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[14]  F. Zito,et al.  Stress response gene activation protects sea urchin embryos exposed to X-rays , 2011, Cell Stress and Chaperones.

[15]  K. Dawson,et al.  Accumulation and embryotoxicity of polystyrene nanoparticles at early stage of development of sea urchin embryos Paracentrotus lividus. , 2014, Environmental science & technology.

[16]  K. Dawson,et al.  Experimental and theoretical comparison of intracellular import of polymeric nanoparticles and small molecules: toward models of uptake kinetics. , 2011, Nanomedicine : nanotechnology, biology, and medicine.

[17]  A. Marcomini,et al.  Titanium dioxide nanoparticles stimulate sea urchin immune cell phagocytic activity involving TLR/p38 MAPK-mediated signalling pathway , 2015, Scientific Reports.

[18]  David Epel,et al.  Embryo stability and vulnerability in an always changing world , 2007, Proceedings of the National Academy of Sciences.

[19]  D. Tocher,et al.  Influence of broodstock diet on somatic growth, fecundity, gonad carotenoids and larval survival of sea urchin , 2015 .

[20]  G. Damonte,et al.  Interactions of cationic polystyrene nanoparticles with marine bivalve hemocytes in a physiological environment: Role of soluble hemolymph proteins. , 2016, Environmental research.

[21]  Fleet David,et al.  Marine Strategy Framework Directive - Task Group 10 Report Marine Litter , 2010 .

[22]  H. Lenihan,et al.  Common strategies and technologies for the ecosafety assessment and design of nanomaterials entering the marine environment. , 2014, ACS nano.

[23]  Luca Monticelli,et al.  Polystyrene Nanoparticles Perturb Lipid Membranes. , 2014, The journal of physical chemistry letters.

[24]  Mark R Wiesner,et al.  Comparison of the abilities of ambient and manufactured nanoparticles to induce cellular toxicity according to an oxidative stress paradigm. , 2006, Nano letters.

[25]  Christine K Payne,et al.  Nanoparticle surface charge mediates the cellular receptors used by protein-nanoparticle complexes. , 2012, The journal of physical chemistry. B.

[26]  M. Wagner,et al.  Characterisation of nanoplastics during the degradation of polystyrene , 2016, Chemosphere.

[27]  J. Evan Ward,et al.  Marine aggregates facilitate ingestion of nanoparticles by suspension-feeding bivalves. , 2009, Marine environmental research.

[28]  Anthony L Andrady,et al.  Microplastics in the marine environment. , 2011, Marine pollution bulletin.

[29]  V. Matranga,et al.  Manganese interferes with calcium, perturbs ERK signaling, and produces embryos with no skeleton. , 2011, Toxicological sciences : an official journal of the Society of Toxicology.

[30]  E. Sarsour,et al.  Manganese superoxide dismutase regulates a redox cycle within the cell cycle. , 2014, Antioxidants & redox signaling.

[31]  Kenneth A Dawson,et al.  Nano-sized polystyrene affects feeding, behavior and physiology of brine shrimp Artemia franciscana larvae. , 2016, Ecotoxicology and environmental safety.

[32]  V. Matranga,et al.  Manganese overload affects p38 MAPK phosphorylation and metalloproteinase activity during sea urchin embryonic development. , 2014, Marine environmental research.

[33]  Isa Doverbratt,et al.  Nanoplastics in the aquatic environment , 2018 .

[34]  R. Burke,et al.  Secondary mesenchyme of the sea urchin embryo: ontogeny of blastocoelar cells. , 1992, The Journal of experimental zoology.

[35]  S. Pokhrel,et al.  Developmental effects of two different copper oxide nanomaterials in sea urchin (Lytechinus pictus) embryos , 2016, Nanotoxicology.

[36]  S. Pokhrel,et al.  Metal oxide nanomaterials in seawater: linking physicochemical characteristics with biological response in sea urchin development. , 2011, Journal of hazardous materials.

[37]  Kou-Juey Wu,et al.  Direct regulation of HSP60 expression by c‐MYC induces transformation , 2008, FEBS letters.

[38]  R. Voellmy.  On mechanisms that control heat shock transcription factor activity in metazoan cells , 2004, Cell stress & chaperones.

[39]  Elenise Gonçalves de Oliveira,et al.  Effects of salinity on the growth, survival, haematological parameters and osmoregulation of tambaqui Colossoma macropomum juveniles , 2015 .

[40]  M. Warnau,et al.  Redox-dependent toxicity of diepoxybutane and mitomycin C in sea urchin embryogenesis. , 2000, Carcinogenesis.

[41]  P. Ramoino,et al.  Developmental abnormalities and changes in cholinesterase activity in sea urchin embryos and larvae from sperm exposed to engineered nanoparticles. , 2013, Aquatic toxicology.

[42]  I. Corsi,et al.  Toxic effects of engineered nanoparticles in the marine environment: model organisms and molecular approaches. , 2012, Marine environmental research.

[43]  Kenneth A. Dawson,et al.  Cationic nanoparticles induce caspase 3-, 7- and 9-mediated cytotoxicity in a human astrocytoma cell line , 2011, Nanotoxicology.

[44]  E. Pelletier,et al.  Toxicity mechanisms of ionic silver and polymer-coated silver nanoparticles with interactions of functionalized carbon nanotubes on early development stages of sea urchin. , 2015, Aquatic toxicology.

[45]  V. Matranga,et al.  Development of a new integrative toxicity index based on an improvement of the sea urchin embryo toxicity test. , 2016, Ecotoxicology and Environmental Safety.

[46]  S. Masters,et al.  14-3-3 proteins: structure, function, and regulation. , 2000, Annual review of pharmacology and toxicology.

[47]  Jeremy C Simpson,et al.  Time resolved study of cell death mechanisms induced by amine-modified polystyrene nanoparticles. , 2013, Nanoscale.

[48]  A. Baun,et al.  Aquatic ecotoxicity testing of nanoplastics - lessons learned from nanoecotoxicology , 2015 .

[49]  Richard C. Thompson,et al.  Microplastic—an emerging contaminant of potential concern? , 2007, Integrated environmental assessment and management.

[50]  Bing Wu,et al.  Copper oxide and zinc oxide nanomaterials act as inhibitors of multidrug resistance transport in sea urchin embryos: their role as chemosensitizers. , 2015, Environmental science & technology.

[51]  J. Lawrence.  Sea urchins : biology and ecology , 2013 .

[52]  C. Bradham,et al.  p38 MAPK in Development and Cancer , 2006, Cell cycle.

[53]  M. Costantini,et al.  Defensome against Toxic Diatom Aldehydes in the Sea Urchin Paracentrotus lividus , 2012, PloS one.

[54]  N. Traverso,et al.  p38MAPK inhibition: a new combined approach to reduce neuroblastoma resistance under etoposide treatment , 2013, Cell Death and Disease.

[55]  F. Zito,et al.  Expression of univin, a TGF-beta growth factor, requires ectoderm-ECM interaction and promotes skeletal growth in the sea urchin embryo. , 2003, Developmental biology.

[56]  Joseph M. DeSimone,et al.  Strategies in the design of nanoparticles for therapeutic applications , 2010, Nature Reviews Drug Discovery.

[57]  V. Matranga,et al.  Sea urchin embryos as an in vivo model for the assessment of manganese toxicity: developmental and stress response effects , 2010, Ecotoxicology.

[58]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[59]  G. Evan,et al.  Caspase‐8 in Apoptosis: The Beginning of “The End”? , 2000, IUBMB life.

[60]  V. Matranga,et al.  Cadmium induces the expression of specific stress proteins in sea urchin embryos. , 2004, Biochemical and biophysical research communications.

[61]  D. Lyons,et al.  Effect of silver nanoparticles on Mediterranean sea urchin embryonal development is species specific and depends on moment of first exposure. , 2015, Marine environmental research.

[62]  A. Gerecke,et al.  Co-release of hexabromocyclododecane (HBCD) and Nano- and microparticles from thermal cutting of polystyrene foams. , 2012, Environmental science & technology.

[63]  I. Corsi,et al.  Particles in the oceans: Implication for a safe marine environment. , 2015, Marine Environmental Research.

[64]  C. Tyler,et al.  Impacts of metal and metal oxide nanoparticles on marine organisms. , 2014, Environmental pollution.

[65]  Richard C. Thompson,et al.  The physical impacts of microplastics on marine organisms: a review. , 2013, Environmental pollution.

[66]  R. Hutz,et al.  Gold Nanoparticles Alter Sea Urchin Development to Pluteus Stage. , 2010 .

[67]  M. Bramini,et al.  Multidisciplinary screening of toxicity induced by silica nanoparticles during sea urchin development. , 2015, Chemosphere.

[68]  J. A. Franeker,et al.  Deleterious Effects of Litter on Marine Life , 2015 .

[69]  M. Roccheri,et al.  Effects of cadmium exposure on sea urchin development assessed by SSH and RT-qPCR: metallothionein genes and their differential induction , 2013, Molecular Biology Reports.