title: Exposure of Paracentrotus lividus male gametes to engineered nanoparticles affects skeletal bio-mineralization processes and larval plasticity.
abstract: The aim of this study is to contribute to the understanding of the mechanisms underlying nanoparticle (NP)-induced embryotoxicity in aquatic organisms. We previously demonstrated that exposure of male gametes to NPs causes non-dose-dependent skeletal damage in sea urchin (Paracentrotus lividus) larvae. In the present study, the molecular mechanisms responsible for these anomalies in sea urchin development from male gametes exposed to cobalt (Co), titanium dioxide (TiO2) and silver (Ag) NPs were investigated by histochemical, immunohistochemical and Western blot analyses. P. lividus sperm were exposed to different NP concentrations (from 0.0001 to 1 mg/L). The distribution of molecules related to skeletogenic cell identification, including ID5 immunoreactivity (IR), wheat germ agglutinin (WGA) affinity and fibronectin (FN) IR, were investigated by confocal laser scanning microscopy at the gastrula (24 h) and pluteus (72 h) stages. Our results identified a spatial correspondence among PMCs, ID5 IR and WGA affinity sites. The altered FN pattern suggests that it is responsible for the altered skeletogenic cell migration, while the Golgi apparatus of the skeletogenic cells, denoted by their WGA affinity, shows different aspects according to the degree of anomalies caused by NP concentrations. The ID5 IR, a specific marker of skeletogenic cells in sea urchin embryos (in particular of the msp130 protein responsible for Ca(2+) and Mg(2+) mineralization), localized in the cellular strands prefiguring the skeletal rods in the gastrula stage and, in the pluteus stage, was visible according to the degree of mineralization of the skeleton. In conclusion, the present study suggests that the investigated NPs suspended in seawater interfere with the bio-mineralization processes in marine organisms, and the results of this study offer a new series of specific endpoints for the mechanistic understanding of NP toxicity.
authors.affiliation: Institute of Marine Science (ISMAR), National Council of Researches (CNR), Via De Marini 6, 16149 Genova, Italy. Electronic address: chiagamba@gmail.com.
authors.forename: Chiara
authors.initials: C
authors.lastname: Gambardella
authors.affiliation: Department of Earth, Environment and Life Sciences (DISTAV), University of Genova, Viale Benedetto XV 5, 16136 Genova, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Ferrando
authors.affiliation: Institute of Marine Science (ISMAR), National Council of Researches (CNR), Via De Marini 6, 16149 Genova, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Morgana
authors.affiliation: Department of Earth, Environment and Life Sciences (DISTAV), University of Genova, Viale Benedetto XV 5, 16136 Genova, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Gallus
authors.affiliation: Department of Earth, Environment and Life Sciences (DISTAV), University of Genova, Viale Benedetto XV 5, 16136 Genova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Ramoino
authors.affiliation: Department of Pharmacy (DIFAR), Biochemistry Lab., University of Genova, Viale Benedetto XV 5, 16136 Genova, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Ravera
authors.affiliation: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia (IIT), Via Morego 30, 16163 Genova, Italy.
authors.forename: Mattia
authors.initials: M
authors.lastname: Bramini
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia (IIT), Via Morego 30, 16163 Genova, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Diaspro
authors.affiliation: Institute of Marine Science (ISMAR), National Council of Researches (CNR), Via De Marini 6, 16149 Genova, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Faimali
authors.affiliation: Department of Earth, Environment and Life Sciences (DISVA), Polytechnic University of Marche, Ancona, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Falugi
mesh_terms: D000818:Animals; D003035:Cobalt / Q000633:toxicity; D004625:Embryo, Nonmammalian / Q000187:drug effects; D007814:Larva / Q000187:drug effects / Q000254:growth & development; D008297:Male; D008670:Metals / Q000633:toxicity*; D053758:Nanoparticles / Q000633:toxicity*; D047328:Paracentrotus / Q000187:drug effects* / Q000254:growth & development; D012834:Silver / Q000633:toxicity; D013094:Spermatozoa / Q000187:drug effects; D014025:Titanium / Q000633:toxicity; D014874:Water Pollutants, Chemical / Q000633:toxicity
chemical_list: D008670:Metals; D014874:Water Pollutants, Chemical; C009495:titanium dioxide; D003035:Cobalt; D012834:Silver; D014025:Titanium
keywords: Biomineralization; Cell migration; Nanoparticles; Paracentrotus lividus; Skeletogenesis; Sperm exposure