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Huang XC, Inoue-Aono Y, Moriyasu Y, et al. Plant Cell Wall-Penetrable, Redox-Responsive Silica Nanoprobe for the Imaging of Starvation-Induced Vesicle Trafficking. Anal Chem. 2016;88(20):10231-10236doi: 10.1021/acs.analchem.6b02920.
Huang, X. C., Inoue-Aono, Y., Moriyasu, Y., Hsieh, P. Y., Tu, W. M., Hsiao, S. C., Jane, W. N., & Hsu, H. Y. (2016). Plant Cell Wall-Penetrable, Redox-Responsive Silica Nanoprobe for the Imaging of Starvation-Induced Vesicle Trafficking. Analytical chemistry, 88(20), 10231-10236. https://doi.org/10.1021/acs.analchem.6b02920
Huang, Xin-Chun, et al. "Plant Cell Wall-Penetrable, Redox-Responsive Silica Nanoprobe for the Imaging of Starvation-Induced Vesicle Trafficking." Analytical chemistry vol. 88,20 (2016): 10231-10236. doi: https://doi.org/10.1021/acs.analchem.6b02920
Huang XC, Inoue-Aono Y, Moriyasu Y, Hsieh PY, Tu WM, Hsiao SC, Jane WN, Hsu HY. Plant Cell Wall-Penetrable, Redox-Responsive Silica Nanoprobe for the Imaging of Starvation-Induced Vesicle Trafficking. Anal Chem. 2016 Oct 18;88(20):10231-10236. doi: 10.1021/acs.analchem.6b02920. Epub 2016 Oct 06. PMID: 27673337.
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Anal Chem. 2016 Oct 18;88(20):10231-10236. doi: 10.1021/acs.analchem.6b02920. Epub 2016 Oct 06.

Plant Cell Wall-Penetrable, Redox-Responsive Silica Nanoprobe for the Imaging of Starvation-Induced Vesicle Trafficking.

Analytical chemistry

Xin-Chun Huang, Yuko Inoue-Aono, Yuji Moriyasu, Pei-Ying Hsieh, Wei-Ming Tu, Shae-Chien Hsiao, Wan-Neng Jane, Hsin-Yun Hsu

Affiliations

  1. Department of Applied Chemistry and Institute of Molecular Science, National Chiao-Tung University , No. 1001 Ta-Hsueh Road, Hsinchu 30010, Taiwan.
  2. Graduate School of Science and Engineering, Saitama University , Shimo-Ohkubo 255, Sakura-Ku, Saitama 338-8570, Japan.
  3. Institute of Plant and Microbial Biology, Academia Sinica , Taipei 11529, Taiwan.

PMID: 27673337 DOI: 10.1021/acs.analchem.6b02920

Abstract

Autophagy is a self-protection process against reactive oxygen species (ROS). The intracellular level of ROS increased when cells were cultured under nutrient starvation. Antioxidants such as glutathione and ascorbic acid play an important role in ROS removal. However, the cellular redox state in the autophagic pathway is still unclear. Herein, we developed a new redox-sensitive probe with a disulfide-linked silica scaffold to enable the sensing of the reduction environment in cell organelles. This redox-responsive silica nanoprobe (ReSiN) could penetrate the plant cell wall and release fluorescent molecules in response to redox states. By applying the ReSiN to tobacco BY-2 cells and tracing the distribution of fluorescence, we found a higher reducing potential in the central vacuole than in the autolysosomes. Upon cysteine protease inhibitor (E64-c) treatment in sucrose-free medium, the disulfide-silica structures of the ReSiNs were broken down in the vacuoles but were not degraded and were accumulated in the autolysosomes. These results reveal the feasibility of our nanoprobe for monitoring the endocytic and macroautophagic pathways. These pathways merge upstream of the central vacuole, which is the final destination of both pathways. In addition, different redox potentials were observed in the autophagic pathway. Finally, the expression of the autophagy-related protein (Atg8) fused with green fluorescence protein confirmed that the ReSiN treatment itself did not induce the autophagic pathway under normal physiological conditions, indicating the versatility of this nanoprobe in studying stimuli-triggered autophagy-related trafficking.

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