title: Detection of malformations in sea urchin plutei exposed to mercuric chloride using different fluorescent techniques.
abstract: Embryos of Mediterranean sea urchin Paracentrotus lividus and subtropical Echinometra mathaei were exposed to 5,10, 15 and 20µgL(-1), and to 1, 2, 3 and 4µgL(-1) mercuric chloride (HgCl2), respectively. The effective concentration (EC50) inducing malformation in 50% of 4-arm pluteus stage (P4) was 16.14µgL(-1) for P. lividus and 2.41µgL(-1) for E. mathaei. Two-photon (TP), second (SHG) and third harmonic generation (THG) microscopy techniques, TUNEL staining, propidium iodide (PI) and Hoechst 33342 probes were used to detect light signals or to stain apoptotic and necrotic cells in fixed and alive plutei. Signals were detected differently in the two species: TP fluorescence, commonly associated with apoptotic cells, did not increase with increasing HgCl2 concentrations in P. lividus and in fact, the TUNEL did not reveal induction of apoptosis. PI fluorescence increased in P. lividus in a dose-dependent manner, suggesting a loss of cell permeability. In E. mathaei plutei TP fluorescence increased at increasing HgCl2 concentrations. THG microscopy revealed skeletal rods in both species. Different fluorescent techniques, used in this study, are proposed as early-warning systems to visualize malformations and physiological responses in sea urchin plutei.
authors.affiliation: Istituto Superiore per la Protezione e la Ricerca Ambientale, ISPRA_ STS-Livorno, Piazzale dei marmi 12, 57123, Italy. Electronic address: isabella.buttino@isprambiente.it.
authors.forename: Isabella
authors.initials: I
authors.lastname: Buttino
authors.affiliation: Institute of Marine Biology, National Taiwan Ocean University, Keelung 20224, Taiwan.
authors.forename: Jiang-Shiou
authors.initials: JS
authors.lastname: Hwang
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Napoli, Italy.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Romano
authors.affiliation: Graduate Institute of Photonics and Optoelectronics and Department of Electrical Engineering, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Chi-Kuang
authors.initials: CK
authors.lastname: Sun
authors.affiliation: Graduate Institute of Photonics and Optoelectronics and Department of Electrical Engineering, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Tzu-Ming
authors.initials: TM
authors.lastname: Liu
authors.affiliation: Istituto Superiore per la Protezione e la Ricerca Ambientale, ISPRA_ STS-Livorno, Piazzale dei marmi 12, 57123, Italy.
authors.forename: David
authors.initials: D
authors.lastname: Pellegrini
authors.affiliation: Istituto Superiore per la Protezione e la Ricerca Ambientale, ISPRA_ STS-Livorno, Piazzale dei marmi 12, 57123, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Gaion
authors.affiliation: Istituto Superiore per la Protezione e la Ricerca Ambientale, ISPRA_ STS-Livorno, Piazzale dei marmi 12, 57123, Italy; CAISIAL, Academic Centre for Innovation and Development in the Food Industry, 80055 Portici (Na), Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Sartori
mesh_terms: D000818:Animals; D017209:Apoptosis; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D008627:Mercuric Chloride / Q000633:toxicity*; D047328:Paracentrotus / Q000187:drug effects* / Q000378:metabolism; D012617:Sea Urchins / Q000187:drug effects* / Q000378:metabolism
chemical_list: D008627:Mercuric Chloride
keywords: Apoptosis; Confocal microscopy; Harmonic generation microscopy; Skeletal rod; Two-photon microscopy