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| Real-time Monitoring of Effects of Two Cytoskeleton Drugs on the Viscoelasticity of Tobacco (Nicotiana tabacum) BY-2 Cells by QCM |
| LAN Ya-Qin1,2, ZHOU Tie-An1,2,*, CHEN Zong-Xing1,2, SU Zhao-Hong2,3, PAN Wei-Song1,2, LIU Yi-Xuan1,2 |
1 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China;
2 Hunan Engineering Research Center for Cell Mechanics and Functional Analysis, Changsha 410128, China;
3 College of Chemistry and Materials Science, Hunan Agricultural University, Changsha 410128, China |
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Abstract Plant cytoskeleton undergoes dynamic remodeling under plant growth and development and different environmental stresses, resulting in changes in the mechanical properties of the cytoskeleton. Cytoskeleton drugs can change the stability of the cytoskeleton, so cytoskeleton drugs can be used to study the viscoelasticity of tobacco (Nicotiana tabacum) BY-2 cells from the level of cellular structure. In this paper, quartz crystal microbalance (QCM) was used to monitor the viscoelastic response of BY-2 cells under the treatments of two cytoskeleton drugs in real time. The results showed that when the cells were treated with low concentrations of paclitaxel (10, 15, 17 μmol/L), the cell viscoelastic index (CVI) of the cells decreased and then increased, and the cells first became softer and then harder. This softening was almost instantaneous, reflecting the rapid force signal participation process. Under the actions of 20 and 25 µmol/L paclitaxel, the CVI of the cells went from negative values to finally close to zero, and the cells became harder and the hardness was greater than that before the drug was added. Under the treatments of lower concentrations of drug cytochalasin D, at low concentrations (50, 100, 150 ng/mL), the CVI of the cells increased from the negative values before adding the drug to greater than zero then dropped, and the cells first became softer and then became harder, the rapid change of CVI's numerical sign reflected the rapid and unstable change of the cells' generated force;and at higher concentrations (200, 250 ng/mL), the CVI of the cells went from negative values to finally slightly greater than zero, and the cells quickly became harder and then became softer, but the hardness was larger than that before adding the drugs. Cellular tensegrity model and the increased plasma membrane tension caused by retraction of the Hechtian strands were used to explain the changes in cells' softness or stiffness under the actions of the two cytoskeleton drugs. In addition, microscopy technology was used to observe the changes in the microfilament structures of tobacco cells under the action of 200 ng/mL cytochalasin D, the result of fluorescence microscopy observation of microfilaments was consistent with the result obtained by QCM real-time monitoring. The method established in this paper provides a new way to study the cellular structure of plant cells based on viscoelasticity measurements, and provides a basis for further research on the mechanical properties of plant cells under the action of cytoskeleton drugs.
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Received: 10 May 2021
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Corresponding Authors:
*tieanzhou@hunau.edu.cn
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