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Assessing the Cytoskeletal System and its Elements in C6 Glioma Cells and Astrocytes by Atomic Force Microscopy

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Abstract

Object To investigate how the characteristic structure of the cytoskeleton in glioma cells is associated with invasiveness. Methods The whole cytoskeletal system was characterized by atomic force microscopy (AFM), while single cytoskeletal elements were exhibited by AFM and using cytoskeletal protein inhibitors to inhibit microfilaments or/and microtubules and displayed by immunofluorescence microscopy. The fluorescence intensity of F-actin was measured by flow cytometry and the structural difference between C6 glioma cells and astrocytes was studied. Results Cytoskeletons in both cells presented network structures, however, the C6 glioma cells showed an irregular edge root and their microfilaments were creber and dense. Intermediate filaments were extensive network structure with non-polarized multipoint connections. The microtubules were relatively big and long and formed tight bundles with close connections between bundles. Astrocytes had a regular and smooth edge, with sparse microfilaments, while the intermediate filaments were dense and interwoven and the microtubules were dense bundled, but only loosely connected each other. Besides, the fluorescence intensity of F-actin was significantly higher in C6 glioma cells (202.54 ± 11.06) than in the astrocytes (62.64 ± 10.23), P < 0.01. Conclusion Whole cytoskeleton and its elements of C6 cells were disclosed of characteristic structures associated with invasiveness. Meanwhile, the content of F-actin could be used as a parameter for measuring cell invasiveness.

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Acknowledgments

The authors thank Dr. Guo Yun-chang from SHIMADIU Japan for AFM technical instruction. This work was supported by National Natural Science Foundation of China (No. 30270491), and also by the Funds for Key Sci-tech Research Projects of Guangdong Province [YUE KEJIBAN (2004) 08, (2007) 05/06-7005206], [YUE CAIQI (2003) 209] of P.R. of China.

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Authors and Affiliations

  1. Neurosurgery Institute of Southern Medical University, Guangzhou, China

    Dexiang Zhou, Xiaodan Jiang, Ruxiang Xu, Yingqian Cai, Jiliang Hu, Gang Xu & Yuxi Zou

  2. Department of Neurosurgery, Zhujiang Hospital of Southern Medical University, Guangzhou, 510282, China

    Dexiang Zhou, Xiaodan Jiang, Ruxiang Xu, Yingqian Cai, Jiliang Hu, Gang Xu & Yuxi Zou

  3. Department of Neurosurgery, the First Affiliated Hospital, Medical College of Shantou University, Shantou, 515041, China

    Dexiang Zhou

  4. Biomechanics & Medical Information Institute, Beijing University of Technology, Beijing, 100022, China

    Yanjun Zeng

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  1. Dexiang Zhou
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  2. Xiaodan Jiang
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  3. Ruxiang Xu
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  4. Yingqian Cai
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  5. Jiliang Hu
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  6. Gang Xu
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  8. Yanjun Zeng
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Correspondence to Xiaodan Jiang.

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Zhou, D., Jiang, X., Xu, R. et al. Assessing the Cytoskeletal System and its Elements in C6 Glioma Cells and Astrocytes by Atomic Force Microscopy. Cell Mol Neurobiol 28, 895–905 (2008). https://doi.org/10.1007/s10571-008-9267-0

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  • DOI: https://doi.org/10.1007/s10571-008-9267-0

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