Decreased Locomotion Behavior Research Articles

Combined Effects of Micro- and Nanoplastics at the Predicted Environmental Concentration on Functional State of Intestinal Barrier in Caenorhabditis elegans.

The possible toxicity caused by nanoplastics or microplastics on organisms has been extensively studied. However, the unavoidably combined effects of nanoplastics and microplastics on organisms, particularly intestinal toxicity, are rarely clear. Here, we employed Caenorhabditis elegans to investigate the combined effects of PS-50 (50 nm nanopolystyrene) and PS-500 (500 nm micropolystyrene) at environmentally relevant concentrations on the functional state of the intestinal barrier. Environmentally, after long-term treatment (4.5 days), coexposure to PS-50 (10 and 15 μg/L) and PS-500 (1 μg/L) resulted in more severe formation of toxicity in decreasing locomotion behavior, in inhibiting brood size, in inducing intestinal ROS production, and in inducting intestinal autofluorescence production, compared with single-exposure to PS-50 (10 and 15 μg/L) or PS-500 (1 μg/L). Additionally, coexposure to PS-50 (15 μg/L) and PS-500 (1 μg/L) remarkably caused an enhancement in intestinal permeability, but no detectable abnormality of intestinal morphology was observed in wild-type nematodes. Lastly, the downregulation of acs-22 or erm-1 expression and the upregulation expressions of genes required for controlling oxidative stress (sod-2, sod-3, isp-1, clk-1, gas-1, and ctl-3) served as a molecular basis to strongly explain the formation of intestinal toxicity caused by coexposure to PS-50 (15 μg/L) and PS-500 (1 μg/L). Our results suggested that combined exposure to microplastics and nanoplastics at the predicted environmental concentration causes intestinal toxicity by affecting the functional state of the intestinal barrier in organisms.

Neuronal Gα subunits required for the control of response to polystyrene nanoparticles in the range of μg/L in C. elegans

The aim of this study was to identify Gα proteins mediating function of neuronal G protein-coupled receptors (GPCRs) in controlling the response to polystyrene nanoparticles (PS-NPs). Caenorhabditis elegans was used as an animal model, and both gene expression and functional analysis were performed to identify the Gα proteins in controlling PS-NPs toxicity. In nematodes, exposure to PS-NPs (1–100 μg/L) significantly altered transcriptional expressions of some neuronal Gα genes, including gpa-5, gpa-10, gpa-11, gpa-15 gsa-1, egl-30, and goa-1. Among these 7 Gα genes, only neuronal RNAi knockdown of gsa-1, gpa-10, and goa-1 affected toxicity of PS-NPs in inducing ROS production and in decreasing locomotion behavior. Some neuronal GPCRs (such as GTR-1, DCAR-1, DOP-2, NPR-8, NPR-12, NPR-9, and DAF-37) functioned upstream of GOA-1, some neuronal GPCRs (such as DCAR-1, DOP-2, NPR-9, NPR-8, and DAF-37) functioned upstream of GSA-1, and some neuronal GPCRs (such as DOP-2, NPR-8, DAF-37, and DCAR-1) functioned upstream of GPA-10 to regulate the toxicity of PS-NPs. Moreover, GOA-1 acted upstream of MPK-1/ERK MAPK, JNK-1/JNK MAPK, DBL-1/TGF-β, and DAF-7/ TGF-β, GSA-1 functioned upstream of MPK-1/ERK MAPK, JNK-1/JNK MAPK, and DBL-1/TGF-β, and GPA-10 functioned upstream of GLB-1/Globin and DBL-1/TGF-β to control the PS-NPs toxicity. Therefore, neuronal Gα proteins of GOA-1, GSA-1, and GPA-10 functioned to transduce signals of multiple GPCRs to different downstream signaling pathways during the control of PS-NPs toxicity in nematodes. Our results provide clues for understanding the important function of GPCRs-Gα signaling cascade in the neurons in controlling response to nanoplastics in organisms.

Dysregulated mir-76 mediated a protective response to nanopolystyrene by modulating heme homeostasis related molecular signaling in nematode Caenorhabditis elegans

The underlying mechanisms of microRNAs (miRNAs) in regulating nanoplastic toxicity are still largely unclear in organisms. In nanopolystyrene (NPS) exposed Caenorhabditis elegans, the expression of mir-76 (a neuronal miRNA) was significantly decreased, and the mir-76 mutant was resistant to the toxicity of NPS. The aim of this study was to determine the molecular basis of mir-76 in controlling NPS toxicity in nematodes. The mir-76 mutation increased expression of glb-10 encoding a globin protein in NPS (1 μg/L) exposed nematodes. Exposure to NPS (1–100 μg/L) increased the glb-10 expression, and the glb-10(RNAi) worm was susceptible to NPS toxicity in inducing reactive oxygen species (ROS) production and in decreasing locomotion behavior. Using ROS production and locomotion behavior as endpoints, mutation of glb-10 inhibited resistance of mir-76 mutant to NPS toxicity, and neuronal overexpression of mir-76 inhibited the resistance to NPS toxicity in nematodes overexpressing neuronal glb-10 containing 3’ untranslated region (3’UTR). Thus, GLB-10 functioned as a target of mir-76 in the neurons to regulate the NPS toxicity. Moreover, a signaling cascade of HRG-7-HRG-5 required for the control of heme homeostasis was identified to function downstream of neuronal GLB-10 to regulate the NPS toxicity. In this signaling cascade, the neuronal HRG-7 regulated the NPS toxicity by antagonizing function of intestinal HRG-5. Furthermore, in the intestine, HRG-5 controlled NPS toxicity by inhibiting functions of hypoxia-inducible transcriptional factor HIF-1 and transcriptional factor ELT-2. Our results highlight the crucial function of heme homeostasis related signaling in regulating the NPS toxicity in organisms.

Induction of protective response to polystyrene nanoparticles associated with methylation regulation in Caenorhabditis elegans

The epigenetic regulation mechanisms for toxicity induction of nanoplastics in organisms remain largely unknown. In Caenorhabditis elegans, we found that prolonged exposure to 1–100 μg/L polystyrene nanoparticles (PS-NPs) decreased expression of MET-2, a H3K9 methyltransferase. Meanwhile, RNAi knockdown of met-2 suppressed the PS-NPs toxicity in inducing production of reactive oxygen species (ROS) and in decreasing locomotion behavior, which suggesting that the decrease in MET-2 expression reflected a protective response. This resistance to PS-NPs toxicity could be further detected in worms with met-2 RNAi knockdown in both intestinal cells and germline cells. In PS-NPs exposed worms, intestinal RNAi knockdown of met-2 significantly increased expressions of daf-16, bar-1, and elt-2. Intestinal RNAi knockdown of daf-16, bar-1, or elt-2 suppressed the resistance of met-2(RNAi) worms to PS-NPs toxicity, suggesting that MET-2 functioned upstream of ELT-2, BAR-1, and DAF-16 in intestinal cells to control PS-NPs toxicity. Moreover, in PS-NPs exposed worms, germline RNAi knockdown of met-2 significantly decreased expressions of wrt-3 and pat-12. RNAi knockdown of wrt-3 or pat-12 further inhibited the susceptibility of worms overexpressing germline MET-2 to PS-NPs toxicity, suggesting that MET-2 functioned upstream of PAT-12 and WRT-3 in germline cells to control PS-NPs toxicity. Therefore, our data provided an important molecular basis for MET-2-mediated methylation regulation in causing protective response to nanoplastics in organisms.

Multigenerational effects of polyethylene terephthalate microfibers in Caenorhabditis elegans

Microfibers (MFs) have recently become an increasingly prevalent pollutant in ecosystems and pose a direct threat to organisms and an indirect threat via adsorption of other pollutants. Here, we used Caenorhabditis elegans to study multigenerational effects of polyethylene terephthalate (PET) MFs (diameter 17.4 μm) by observing the maternal generation (P0) to the seventh offspring generation (F7) with continuous MF exposure. Exposure to 250-μm PET MFs decreased locomotion behavior and induced intestinal reactive oxygen species (ROS) in the P0 generation compared with other PET MF sizes. Moreover, no notably negative effects on survival were observed in any generation during continuous exposure to 250-μm PET MFs. However, the reproduction rate clearly decreased in the F2 and F3 generations but gradually recovered in the F4–F7 generations. Developmental abnormalities showed a close relationship with body length. Although some recovery was confirmed, there were significant decreases in body length in the F2–F5 generations. Interestingly, growth inhibition was also observed in the F6 generation without MF exposure. ROS production and dermal damage in the P0–F5 generations might have resulted in the toxicological responses. To the best of our knowledge, this is the first study to provide evidence of multigenerational toxicity of MFs in C. elegans.

Effect of chronic exposure to nanopolystyrene on nematode Caenorhabditis elegans

Nanoplastic exposure could cause toxicity to Caenorhabditis elegans at various aspects. Nevertheless, the effects of chronic exposure to nanoplastics remain largely unclear in nematodes. In this study, we employed C. elegans as an animal model to determine the effects of nanopolystyrene (30 nm) exposure from adult day-1 for 8-day. After the exposure, only 1000 μg/L nanopolystyrene reduced the lifespan. In contrast, nanopolystyrene ≥1 μg/L decreased locomotion behavior and activated oxidative stress. Meanwhile, in 10 μg/L nanopolystyrene exposed nematodes, both expression of SOD-3, a Mn-SOD, and autophagy induction as indicated by LGG-1:GFP expression were significantly increased. RNAi knockdown of daf-2 encoding an insulin receptor enhanced the autophagy induction, and RNAi knockdown of daf-16 encoding a FOXO transcriptional factor in insulin signaling pathway suppressed the autophagy induction in 10 μg/L nanopolystyrene exposed nematodes. Moreover, DAF-16 acted upstream of LGG-1, an ortholog of Atg8/LC3, to regulate the toxicity of nanopolystyrene toxicity in inducing ROS production and in decreasing locomotion behavior at adult day-9. Our data implied the potential toxicity of chronic exposure to nanoplastics at predicted environmental concentrations on organisms.

Potential of esterase DmtH in transforming plastic additive dimethyl terephthalate to less toxic mono-methyl terephthalate

Dimethyl terephthalate (DMT) is a primary ingredient widely used in the manufacture of polyesters and industrial plastics; its environmental fate is of concern due to its global use. Microorganisms play key roles in the dissipation of DMT from the environment; however, the enzymes responsible for the initial transformation of DMT and the possible altered toxicity due to this biotransformation have not been extensively studied. To reduce DMT toxicity, we identified the esterase gene dmtH involved in the initial transformation of DMT from the AOPP herbicide-transforming strain Sphingobium sp. C3. DmtH shows 24–41% identity with α/β-hydrolases and belongs to subfamily V of bacterial esterases. The purified recombinant DmtH was capable of transforming DMT to mono-methyl terephthalate (MMT) and potentially transforming other p-phthalic acid esters, including diallyl terephthalate (DAT) and diethyl terephthalate (DET). Using C. elegans as an assay model, we observed the severe toxicity of DMT in inducing reactive oxygen species (ROS) production, decreasing locomotion behavior, reducing lifespan, altering molecular basis for oxidative stress, and inducing mitochondrial stress. In contrast, exposure to MMT did not cause obvious toxicity, induce oxidative stress, and activate mitochondrial stress in nematodes. Our study highlights the usefulness of Sphingobium sp. C3 and its esterase DmtH in transforming p-phthalic acid esters and reducing the toxicity of DMT to organisms.

Biosafety assessment of water samples from Wanzhou watershed of Yangtze Three Gorges Reservoir in the quiet season in Caenorhabditis elegans

We here employed a model animal of Caenorhabditis elegans to perform toxicity assessment of original surface water samples collected from Three Gorges Reservoir (TGR) in the quiet season in Wanzhou, Chongqing. Using some sublethal endpoints, including lifespan, body length, locomotion behavior, brood size, and intestinal reactive oxygen species (ROS) induction, we found that the examined five original surface water samples could not cause toxicity on wild-type nematodes. Nevertheless, the surface water sample collected from backwater area induced the significant increase in expressions of genes (sod-2 and sod-3) encoding Mn-SODs in wild-type nematodes. Among the examined five original surface water samples, exposure to the original surface water sample collected from backwater area could further cause the toxicity in decreasing locomotion behavior and in inducing intestinal ROS production in sod-3 mutant nematodes. Moreover, the solid phase of surface water sample collected from backwater area might mainly contribute to the observed toxicity in sod-3 mutant nematodes. Our results are helpful for understanding the potential effects of surface water in the TGR region in the quiet season on environmental organisms.

Combinational effect of titanium dioxide nanoparticles and nanopolystyrene particles at environmentally relevant concentrations on nematode Caenorhabditis elegans

The possible adverse effects of nanoplastics have received the great attention recently; however, their effects at environmentally relevant concentration on organisms are still largely unclear. We here employed Caenorhabditis elegans to investigate the combinational effects of titanium dioxide nanoparticles (TiO2-NPs) and nanopolystyrene particles at environmentally relevant concentrations on organisms. In wild-type nematodes, prolonged exposure to nanopolystyrene particles (1 μg/L) could enhance the toxicity of TiO2-NPs (1 μg/L) in decreasing locomotion behavior and in inducing intestinal reactive oxygen species (ROS) production. Meanwhile, combinational exposure to TiO2-NPs (1 μg/L) and nanopolystyrene particles (1 μg/L) altered the molecular basis for oxidative stress in wild-type nematodes. Moreover, prolonged exposure to nanopolystyrene particles (0.1 μg/L) could further enhance the toxicity of TiO2-NPs (1 μg/L) in decreasing locomotion behavior and in inducing intestinal ROS production in sod-3 mutant nematodes. Our data suggest the potential role of nanopolystyrene particles at environmentally relevant concentrations in enhancing the toxicity of ENMs in the environment.

Metallothioneins act downstream of insulin signaling to regulate toxicity of outdoor fine particulate matter (PM2.5) during Spring Festival in Beijing in nematode Caenorhabditis elegans.

In this study, we performed the toxicological assessment of outdoor PM2.5 collected from Beijing during Spring Festival using the in vivo assay system of Caenorhabditis elegans. Acute exposure to outdoor PM2.5 at a concentration of 10 mg L-1 and prolonged exposure to outdoor PM2.5 at concentrations of 0.1-10 mg L-1 decreased locomotion behavior and caused significant induction of intestinal ROS production. Meanwhile, outdoor PM2.5 exposure induced significant expression of gene (mtl-1 and mtl-2) encoded metallothioneins in the intestine. Mutation of the mtl-1 or mtl-2 gene resulted in a susceptible property of nematodes to outdoor PM2.5 toxicity. Genetic assays suggested that mtl-1 and mtl-2 genes acted downstream of the daf-16 gene encoding a FOXO transcriptional factor and daf-2 gene encoding an insulin receptor in the insulin signaling pathway to regulate outdoor PM2.5 toxicity. DAF-2 further acted upstream of DAF-16 and suppressed the function of DAF-16 to regulate outdoor PM2.5 toxicity. Therefore, we identified a signaling cascade of DAF-2-DAF-16-MTL-1/2 in the control of outdoor PM2.5 toxicity in nematodes. Our study provides an important molecular basis for the potential toxicity of outdoor PM2.5 during Spring Festival in Beijing in nematodes. Especially, our study will highlight the potential adverse effects of outdoor PM2.5 during Spring Festival on environmental organisms.