Abstract

Effects of ZnO NPs and ionic Zn on germline apoptosis and the regulation of genes in the apoptosis pathway were investigated in vivo using the model organism Caenorhabditis elegans.Age synchronized Bristol N2 worms were exposed to ZnO NPs and ZnCl2 at concentrations of 6.14 × 10-1, 61.4, and 614 μM form larval stage 1 (L1) to early adulthood. Possible ZnO nanoparticles were observed under the worm cuticle and also in the gonadal region by transmission electron microscopy (TEM). ZnO NPs and ZnCl2 both significantly increased the number of apoptotic cells as compared with controls in the 61.4 and 614 μM treatment groups (P < .05). However, ZnO NPs induced more apoptotic cells in the 61.4 μM treatment than ZnCl2 (P < .05), suggesting ZnO NP is more potent in inducing apoptosis at specific exposure concentration. Findings using the MD701 (bcIs39 [(lim-7)ced-1p::GFP + lin-15(+)]) strain further confirmed the observations in N2 strain. Genes involved in the apoptosis pathway (ced-13, ced-3, ced-4, ced-9, cep-1, dpl-1, efl-1, efl-2, egl-1, egl-38, lin-35, pax-2, and sir-2.1) were in general upregulated in response to ZnO NP exposure. The cep-1/p53 gene was up-regulated in gene expression assay. In the cep-1 loss of function mutant, no significant increase in apoptosis was observed. Therefore, the increased apoptosis resulting from ZnO NPs exposure is likely cep-1/p53 dependent. This study provides evidence that ZnO nanoparticles affect germ cell apoptotic machinery as a potential mechanism of reproductive toxicity.

Highlights

  • Nanomaterials are produced in metric tons per year and the production is expected to increase over the decade (Nel et al, 2006; USEPA, 2005)

  • Effects of ZnO NPs and ionic Zn on germline apoptosis and the regulation of genes in the apoptosis pathway were investigated in vivo using the model organism Caenorhabditis elegans.Age synchronized Bristol N2 worms were exposed to ZnO NPs and ZnCl2 at concentrations of 6.14 Â 10À1, 61.4, and 614 lM form larval stage 1 (L1) to early adulthood

  • This study investigated effects of ZnO NPs exposure on C. elegans germ cell apoptosis and related gene expressions, proposed as a potential mechanism of observed reproductive inhibition reported by previous studies

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Summary

Introduction

Nanomaterials are produced in metric tons per year and the production is expected to increase over the decade (Nel et al, 2006; USEPA, 2005). Previous studies have used the model organism Caenorhabditis elegans to assess effects of manufactured ZnO. It was found that ZnO NPs exposure results in inhibition of growth, behaviors, reproduction or increased mortality at various concentrations including environmentally relevant concentrations (Ma et al, 2009; Wang et al, 2009; Wu et al, 2013). This study investigated effects of ZnO NPs exposure on C. elegans germ cell apoptosis and related gene expressions, proposed as a potential mechanism of observed reproductive inhibition reported by previous studies. Since dissolved ionic Zn may contribute to the toxic effects, we are interested in comparing the apoptosis-inducing effects of ZnO NPs with ZnCl2

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