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Three-dimensional fine structure of the organization of microtubules in neurite varicosities by ultra-high voltage electron microscope tomography

Cell and Tissue Research volume 369, pages 467–476 (2017)Cite this article

Abstract

Neurite varicosities are highly specialized compartments that are involved in neurotransmitter/ neuromodulator release and provide a physiological platform for neural functions. However, it remains unclear how microtubule organization contributes to the form of varicosity. Here, we examine the three-dimensional structure of microtubules in varicosities of a differentiated PC12 neural cell line using ultra-high voltage electron microscope tomography. Three-dimensional imaging showed that a part of the varicosities contained an accumulation of organelles that were separated from parallel microtubule arrays. Further detailed analysis using serial sections and whole-mount tomography revealed microtubules running in a spindle shape of swelling in some other types of varicosities. These electron tomographic results showed that the structural diversity and heterogeneity of microtubule organization supported the form of varicosities, suggesting that a different distribution pattern of microtubules in varicosities is crucial to the regulation of varicosities development.

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Acknowledgment

This work was supported by the “Nanotechnology Platform” (project No. 12024046) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.

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Author notes

  1. Tomoki Nishida

    Present address: Japan Textile Products Quality and Technology Center, 5-7-3, Shimoyamate St, Chuo-ku, Kobe, Hyogo, 650-0011, Japan

Authors and Affiliations

  1. Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, 7-1, Mihogaoka, Osaka, Ibaraki, 567-0047, Japan

    Tomoki Nishida

  2. Department of Applied Biology, Graduate School of Science and Technology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, 606-8585, Japan

    Ryoichi Yoshimura & Yasuhisa Endo

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  1. Tomoki Nishida
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  2. Ryoichi Yoshimura
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  3. Yasuhisa Endo
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Corresponding author

Correspondence to Tomoki Nishida.

Electronic supplementary material

ESM 1

Three-dimensional models of cytoskeletons and membrane organelles in the neurite varicosity. Microtubules, neurofilaments, clathrin-coated vesicles, endosome and mitochondria are colored green, purple, blue, yellow and white, respectively. Scale bar, 500 nm. (related to Fig. 2 and S1) (AVI 12881 kb)

ESM 2

Three-dimensional models of microtubules and membrane organelles in a small cytoplasmic expansion at the neurite shaft. Microtubules, clathrin-coated vesicles, endosome and plasma membrane are in green, blue, yellow and white respectively. A part of the plasma membrane of varicosity and swollen areas are colored red and purple, respectively. Scale bar, 500 nm. (related to Fig. 2 and S2) (AVI 12282 kb)

ESM 3

Three-dimensional models of microtubules and mitochondrion in a beaded varicosity that was reconstructed from two serial tomograms. Microtubules, spherical structures of microtubule tip and mitochondrion are in green, yellow and purple, respectively. Scale bar, 500 nm. (related to Fig. 3 and S3) (AVI 9363 kb)

ESM 4

Three-dimensional models of microtubule distribution in a globular varicosity branch that was reconstructed from four serial tomograms. Microtubules, looped microtubule, spherical structures of microtubule tip, plasma membrane and clathrin-coated vesicles are colored green, red, yellow, white and blue, respectively. Scale bar, 500 nm. (related to Fig. 3 and S4) (AVI 12484 kb)

ESM 5

Three-dimensional models of microtubules in a swelled neurite branching site that was reconstructed from two serial tomograms. Microtubules, spherical structures of microtubule tip, looped microtubule, two kinds of large circular turn microtubules and mitochondria are colored green, yellow, red, blue, yellow and white, respectively. Scale bar, 500 nm. (related to Fig. 4 and S5) (AVI 10112 kb)

ESM 6

Three-dimensional models of microtubules in a detergent-extracted varicosity. Microtubules, looped microtubule and spherical structures of microtubule tip are colored green, red and yellow, respectively. Scale bar, 500 nm. (related to Fig. 5 and S6) (AVI 11289 kb)

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Nishida, T., Yoshimura, R. & Endo, Y. Three-dimensional fine structure of the organization of microtubules in neurite varicosities by ultra-high voltage electron microscope tomography. Cell Tissue Res 369, 467–476 (2017). https://doi.org/10.1007/s00441-017-2645-5

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  • DOI: https://doi.org/10.1007/s00441-017-2645-5

Keywords

  • Varicosity
  • Microtubule
  • Neurite
  • Electron tomography
  • Ultra-high voltage electron microscope