Impact of copper oxide nanoparticles (CuO NPs) exposure on embryo development and expression of genes related to the innate immune system of zebrafish (Danio rerio).

CuO NPs are nanomaterials with catalytic activity and unique thermo-physical properties used in different fields such as sensors, catalysts, surfactants, batteries, antimicrobials and solar energy transformations. Because of its wide field of use, these nanoparticles accumulate in the aquatic environment and thus lead to toxic effects on aquatic organisms. The toxicological findings about CuO NPs are controversial and these effects of CuO NPs on aquatic organisms have not been elucidated in detail. Therefore, the aim of this study was to investigate the toxic effect of CuO NPs on zebrafish embryos using different parameters including molecular and morphologic. For this purpose, zebrafish embryos at 4 h after post fertilization (hpf) were exposed to different concentrations of CuO NPs (0.5, 1, 1.5 mg/L) until 96 hpf. Mortality, hatching, heartbeat, malformation rates were examined during the exposure period. In addition, Raman spectroscopy was used to determine whether CuO NPs entered into the tissues of zebrafish larvae or not. Moreover, the alterations in the expression of genes related to the antioxidant system and innate immune system were examined in the embryos exposed to CuO NPs during 96 h. The results showed that CuO NPs was not able to enter into the zebrafish embryos/larvae tissues but caused an increased the mortality rate, a delayed hatching, and a decreased heartbeat rate. Moreover, CuO NPs caused several types of abnormalities such as head and tail malformations, vertebral deformities, yolk sac edema, and pericardial edema. RT-PCR results showed that the transcription of mtf-1, hsp70, nfkb and il-1β, tlr-4, tlr-22, trf, cebp was changed by the application of CuO NPs. In conclusion, short-term exposure to CuO NPs has toxic effects on the development of zebrafish embryos.

Aksakal FI ，Ciltas A 《COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY》

A holistic study on potential toxic effects of carboxylated multi-walled carbon nanotubes (MWCNTs-COOH) on zebrafish (Danio rerio) embryos/larvae.

Multi-walled carbon nanotubes (MWCNTs) have widespread use in industrial and consumer products and great potential in biomedical applications. This leads to inevitably their release into the environment and the formation of their toxic effects on organisms. These effects can change depending on their physicochemical characteristics. Therefore, the toxicological findings of MWCNTs are inconsistent. Their toxicities related to surface modification have not been elucidated in a holistic manner. Hence, this study was conducted to clarify their potential toxic effects on zebrafish embryos/larvae in a comprehensive approach using morphologic, biochemical and molecular parameters. Zebrafish embryos were exposed to 5, 10, 20 mg/L doses of MWCNTs-COOH at 4 h after fertilization and grown until 96 hpf. Physiological findings demonstrated that they induced a concentration-dependent increase in the mortality rate, delayed hatching and decrease in the heartbeat rate. Moreover, it caused abnormalities including yolk sac edema, pericardial edema, head, tail malformations, and vertebral deformities. These effects may be due to the alterations in antioxidant and immune system related gene expressions after their entry into zebrafish embryo/larvae. The entry was confirmed from the evaluation of Raman spectra collected from the head, yolk sac, and tail of control and the nanotube treated groups. The gene expression analysis indicated the changes in the expression of oxidative stress (mtf-1, hsp70, and nfkb) and innate immune system (il-1β, tlr-4, tlr-22, trf, and cebp) related genes, especially an increased in the expression of the hsp70 and il-1β. These findings proved the developmental toxicities of MWCNTs-COOH on the zebrafish embryos/larvae.

Icoglu Aksakal F ，Ciltas A ，Simsek Ozek N 《-》

Developmental toxicity induced by Cu(OH)(2) nanopesticide in zebrafish embryos.

The current study evaluates the adverse effects of Cu(OH)2 nanopesticide (CNPE) on the early life stages of zebrafish (Danio rerio). The developmental toxicity was determined using different parameters such as mortality (including LC50 ), hatching, heart rates, malformations, and alteration of the gene expressions. Zebrafish embryos (4 hpf-hours postfertilization) were exposed to 1.0, 2.0, 4.0, 8.0, and 16.0 mg/l CNPE doses until 96 hpf. The 96 hours LC50 was recorded at 6.258 mg/l. Seventy-two hpf total malformation index values for 2.0, 4.0, and 8.0 mg/l CNPE doses were 4.3, 7.2 and 7.9, respectively. 1.0 mg/l CNPE is not toxic for the zebrafish embryos/larvae. 2.0 to 8.0 CNPE doses caused some abnormalities in embryos/larvae morphology, including lack of body parts, tail deformities, chorda deformity, bubbled head, scoliosis, lordosis, weak or non-pigmentation, decreased heart rate and larva length. 16.0 mg/l CNPE caused mortality in 72 hpf. The expression levels of seven immune system-related genes (il-1β, il-8, cebp, tlr4, hsp70, NF-kB, and mtf-1) were examined. The transcription level of il-1β, il-8, tlr4, hsp70, and NF-kB genes significantly increased in the CNPE exposure groups. While the expression of the mtf-1 gene considerably decreased, the cebp gene expression level did not change in the 4.0 and 8.0 mg/l CNPE doses. In conclusion, CNPE could induce developmental toxicity with malformations in embryos/larvae and alter the gene expression.

Aksakal FI ，Sisman T 《-》

Montmorillonite clay and humic acid modulate the behavior of copper oxide nanoparticles in aqueous environment and induces developmental defects in zebrafish embryo.

Copper oxide nanoparticles (CuO NPs) is one of the most commonly used metal oxide nanoparticles for commercial and industrial products. An increase in the manufacturing and use of the CuO NPs based products has increased the likelihood of their release into the aquatic environment. This has attracted major attention among researchers to explore their impact in human as well as environmental systems. CuO NPs, once released into the environment interact with the biotic and abiotic constituents of the ecosystem. Hence the objective of the study was to provide a holistic understanding of the effect of abiotic factors on the stability and aggregation of CuO NPs and its correlation with their effect on the development of zebrafish embryo. It has been observed that the bioavailability of CuO NPs decrease in presence of humic acid (HA) and heteroagglomeration of CuO NPs occurs with clay minerals. CuO NPs, CuO NPs + HA and CuO NPs + Clay significantly altered the expression of genes involved in development of dorsoventral axis and neural network of zebrafish embryos. However, the presence of HA with clay showed protective effect on zebrafish embryo development. These findings provide new insights into the interaction of NPs with abiotic factors and combined effects of such complexes on developing zebrafish embryos genetic markers.

Kansara K ，Paruthi A ，Misra SK ，Karakoti AS ，Kumar A ... -  《-》

The effects of copper oxide nanoparticles on dorsoventral patterning, convergent extension, and neural and cardiac development of zebrafish.

Currently, nanoparticles are widely used in biomedicine and industry. CuO nanoparticles (CuO-NPs) are versatile materials in our daily life and their toxicity has drawn extensive attention. In this study, we concentrate on the effect of CuO-NPs on early zebrafish development. The results reveal that CuO-NPs can induce abnormal phenotypes of a smaller head and eyes and delayed epiboly. The gene expression pattern shows that CuO-NPs spatially narrow the expression of dorsal genes chordin and goosecoid and alter the expression of dlx3, ntl and hgg which are related to the cell migration of gastrulation. The decreased expression of pax2 and pax6 involved in neural differentiation was accordant with the decreased sizes of neural structures. Cmlc2 expression suggests that CuO-NPs prevented looping of the heart tube during cardiogenesis. Furthermore, quantitative RT-PCR results suggest that the CuO-NPs could increase the canonical Wnt signaling pathway to narrow the expression of chordin and goosecoid in dorsoventral patterning as well as decrease the transcription of Wnt5 and Wnt11 to result in slower, less directed movements and an abnormal cell shape. These findings indicated the CuO-NPs exert developmental toxicity. The present study evaluates the ecological and developmental toxicity, providing warnings about the application of CuO-NPs.

Xu J ，Zhang Q ，Li X ，Zhan S ，Wang L ，Chen D ... -  《-》