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Effects of Titanium Dioxide Nanoparticles on the Synthesis of Fibroin in Silkworm (Bombyx mori)

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Abstract

Silkworm (Bombyx mori) is an economically important insect, and its silk production capacity largely depends on its ability to synthesize fibroin. While breeding of B. mori varieties has been a key strategy to improve silk production, little improvement of B. mori silk production has been achieved to date. As a result, the development of sericulture economy has not progressed well, pointing to the need of new ways for improvement of B. mori silk production. Titanium dioxide nanoparticles (TiO2 NPs), a food additive widely used for livestock, have been shown to promote animal growth and increase the protein synthesis in animals. However, no studies on effect of TiO2 NPs on fibroin synthesis in B. mori have been available. In this study, the differential expression profiles of genes and proteins in the silk gland of B. mori fed without or with TiO2 NPs (5 μg ml−1) were analyzed and compared using digital gene expression (DGE), reverse transcription quantitative polymerase chain reaction (RT-qPCR), semi-qPCR, and Western blot analysis. The effects of TiO2 NPs feeding on the activity of proteases in the midgut and the synthesis and transportation of amino acids in hemolymph were also investigated. DGE analyses showed that among a total of 4,741 genes detected, 306 genes were differentially expressed after the TiO2 NPs feeding, of which 137 genes were upregulated whereas 169 genes were downregulated. 106 genes were shown to be involved in fibroin synthesis, of which 97 genes, including those encoding cuticular protein glycine-rich 10, serine protease inhibitor 28, aspartate aminotransferase, lysyl-tRNA synthetase, and splicing factor arginine/serine-rich 6, and silk gland factor-1 (SGF-1), were upregulated with the maximum induction of 8.52-folds, whereas nine genes, including those encoding aspartylglucosaminidase, the cathepsin L in Tribolium castaneum, and similar to SPRY domain-containing SOCS box protein 3, were downregulated with the maximum reduction of 8.11-folds. Transcription levels of nine genes were further verified by RT-qPCR, and the results were consistent with those with DGE. Transcription and expression levels of fibroin light chain (Fib-L) gene were increased after TiO2 NPs feeding, indicating that TiO2 NPs improves fibroin synthesis. Compared with that of control, the mean protease activity was increased by 56.67 % in the B. mori fed with TiO2 NPs, and the transport of four key amino acids used for fibroin synthesis in hemolymph was also increased. These findings indicated that TiO2 NPs feeding can improve the absorption and utilization of amino acids from the feed and could be a new way to increase the fibroin synthesis in B. mori.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program; Grant no. 2013AA102507), and the projects were sponsored by the National Cocoons Silk Development Funds in 2014, the Priority Academic Program Development of Jiangsu Higher Education Institutions, the Doctoral Fund of Ministry of Education of China (20113201110008), the China Agriculture Research System (CARS-22-ZJ0305), the Science and Technology Support Program of Suzhou (ZXS2012005, SYN201406), and the Graduate Education Innovation Project of Jiangsu Province (CXLX13_821, CXZZ13_0821).

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Authors and Affiliations

  1. School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, 215123, People’s Republic of China

    Min Ni, FanChi Li, JiangHai Tian, JingSheng Hu, Hua Zhang, KaiZun Xu, BinBin Wang, YangYang Li, WeiDe Shen & Bing Li

  2. National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu, 215123, People’s Republic of China

    WeiDe Shen & Bing Li

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  1. Min Ni
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  2. FanChi Li
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  3. JiangHai Tian
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  4. JingSheng Hu
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  5. Hua Zhang
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  10. Bing Li
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Correspondence to Bing Li.

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Min Ni, FanChi Li, and JiangHai Tian contributed equally to the study.

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Table S1

DGE data analysis for posterior silk gland of B. mori fed with the TiO2 NPs (DOC 182 kb)

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Ni, M., Li, F., Tian, J. et al. Effects of Titanium Dioxide Nanoparticles on the Synthesis of Fibroin in Silkworm (Bombyx mori). Biol Trace Elem Res 166, 225–235 (2015). https://doi.org/10.1007/s12011-015-0258-y

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