Maternal exposure to urban particulate matter induces cardiac developmental toxicity in zebrafish offspring by disrupting mitochondrial homeostasis

Figures

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  Fig. 1. Reproductive toxicity in female zebrafish caused by UPM exposure. A. The representative images of H&E staining in ovary sections (scale bar: 200 μm). n = 5. a. primary growth oocyte; b. cortical follicular oocyte; c. vitellogenic oocyte; d. postovulatory follicle. B. The number of zebrafish embryos. n = 14–15. C. The fertilization rate of zebrafish embryos. n = 14–15. D. The representative optical images of deformed zebrafish at 48 and 72 hpf (scale bar: 200 μm). n = 6. Red arrow: pericardial edema. Blue arrow: spinal curvature. *p < 0.05, compared to the control group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

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  Fig. 2. Abnormal heart development in offspring zebrafish after maternal UPM exposure. A-B. The representative optical images of cardiotoxicity phenotype at 72 hpf (scale bar: 200 μm) and quantitative analysis of pericardium edema area. n = 6. C. The heart rate of zebrafish embryos at 72 hpf. n = 28–30. D. The mRNA expression of Nkx2.5, Tbx5, Vmhc, Myh6, Tbx2b and Notch1b. n = 5. E. The representative images of H&E staining in offspring zebrafish at 7 and 14 dpf (scale bar: 20 μm). n = 6. Red arrow: inflammatory cells. A: atrium. V: ventricle. *p < 0.05, compared to the control group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

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  Fig. 3. Transcriptomics analysis of zebrafish offspring. A. Clustering map of differentially expressed genes. B. Up-regulation and down-regulation significant pathways. C. Statistics of GO enrichment for genes. D. Clustering of pathways on top pathways. E. Clustering of pathways on middle pathways.

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  Fig. 4. Results of WGCNA. A. Network heatmap plot. B. Hierarchical clustering dynamic tree diagram. C. Analysis of the scale-free index for various soft-threshold powers. D. Analysis of the mean connectivity for various soft-threshold powers. E. Module-trait relationship heat map. F. Eigengene adjacency heatmap. G. WGCNA network. H. Gene-disease association network.

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  Fig. 5. Mitochondria-related dysfunctions in offspring zebrafish following maternal UPM exposure. A. Heatmap of key genes clustering. B. qRT-PCR analysis. n = 5. C. The mRNA expression of Fis1 and Opa1. n = 5. D. Curves of oxygen consumption rate. E. The quantitative analysis of mitochondrial respiration. n = 6. *p < 0.05, compared to the control group.

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  Fig. 6. ANGPTL4 plays a critical role in UPM-induced abnormal heart development in offspring. A. The representative immunofluorescence images of ANGPTL4 (scale bar: 20 μm). B-C. The representative immunofluorescence images of Fis1 (scale bar: 10 μm). D-E. The representative immunofluorescence images of Opa1 (scale bar: 10 μm). F. The quantitative analysis of ANGPTL4 fluorescence intensity. n = 5. G. The cell viability of AC16 cells. n = 5. H-I. The quantitative analysis of Fis1 fluorescence intensity. n = 5. J-K. The quantitative analysis of Opa1 fluorescence intensity. n = 5. L-M. The quantitative analysis of MMP level by flow cytometry. n = 5. N-O. The quantitative analysis of mtROS level by flow cytometry. n = 5. *p < 0.05, compared to the control group. #p < 0.05, compared to the UPM group.