Zebrafish High-Throughput Screening to Study the Impact of Dissolvable Metal Oxide Nanoparticles on the Hatching Enzyme, ZHE1

• Published in: Small (Weinheim an der Bergstrasse, Germany) Vol. 9; no. 9-10; pp. 1776 - 1785
• Main Authors: Lin, Sijie, Zhao, Yan, Ji, Zhaoxia, Ear, Jason, Chang, Chong Hyun, Zhang, Haiyuan, Low-Kam, Cecile, Yamada, Kristin, Meng, Huan, Wang, Xiang, Liu, Rong, Pokhrel, Suman, Mädler, Lutz, Damoiseaux, Robert, Xia, Tian, Godwin, Hilary A., Lin, Shuo, Nel, André E.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 27.05.2013; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: Amino Acid Sequence; Animals; COPPER OXIDE; CUPRIC OXIDE; Danio rerio; Embryos; Enzymes; Freshwater; hatching interference; high throughput screening; High-Throughput Screening Assays; IMPACT PROPERTIES; Interference; Metal Nanoparticles - chemistry; Metal Nanoparticles - toxicity; Metal oxides; Metalloproteases - metabolism; Molecular Sequence Data; Nanoparticles; Nanotechnology; OXIDES; Oxides - chemistry; PARTICLES; predictive toxicology; Proteins; ROBOTS; Screening; Solubility; Zebrafish; Zebrafish - embryology
• ISSN: 1613-6810; 1613-6829
• DOI: 10.1002/smll.201202128

The zebrafish is emerging as a model organism for the safety assessment and hazard ranking of engineered nanomaterials. In this Communication, the implementation of a roboticized high‐throughput screening (HTS) platform with automated image analysis is demonstrated to assess the impact of dissolvable oxide nanoparticles on embryo hatching. It is further demonstrated that this hatching interference is mechanistically linked to an effect on the metalloprotease, ZHE 1, which is responsible for degradation of the chorionic membrane. The data indicate that 4 of 24 metal oxide nanoparticles (CuO, ZnO, Cr2O3, and NiO) could interfere with embryo hatching by a chelator‐sensitive mechanism that involves ligation of critical histidines in the ZHE1 center by the shed metal ions. A recombinant ZHE1 enzymatic assay is established to demonstrate that the dialysates from the same materials responsible for hatching interference also inhibit ZHE1 activity in a dose‐dependent fashion. A peptide‐based BLAST search identifies several additional aquatic species that express enzymes with homologous histidine‐based catalytic centers, suggesting that the ZHE1 mechanistic paradigm could be used to predict the toxicity of a large number of oxide nanoparticles that pose a hazard to aquatic species. Automated high‐throughput screening using zebrafish embryos is used for hazard assessment of 24 representative metal oxide nanoparticles. Four nanoparticles are found to interfere with zebrafish embryo hatching. Hatching interference is a result of toxic metal ion shedding from nanoparticles, compromising the zebrafish hatching enzyme 1 (ZHE1) activity. The structural and functional similarities of hatching enzymes across fish species suggest that the ZHE1 mechanistic paradigm could be used to predict the toxicity of a large number of engineered nanoparticles.