Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Yoichiro Sogame; Katsuhiko Kojima; Toshikazu Takeshita; Shiho Kikuchi; Yuto Shimada; Rikiya Nakamura; Mikihiko Arikawa; Seiji Miyata; Eiji Kinoshita; Futoshi Suizu; Tatsuomi Matsuoka

Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Publication date: 17.03.2021

Acta Protozoologica, 2020, Volume 59, Issue 3-4, pp. 107 - 120

https://doi.org/10.4467/16890027AP.20.009.13264

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Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Abstract

Assays of protein contained in water-soluble fraction of encysting cells Colpoda cucullus Nag-1 by two-dimensional electrophoresis (2-D PAGE) and mass spectrometry (MS) revealed that the amount of β-tubulin abruptly increased in 2.5–10 h after encystment induction. Judging from the results that total α-tubulin content did not decrease much until 12 h after encystment induction, the result indicates that disassembly of microtubules may occur soon after encystment is induced. Therefore, we tried to visualize dynamics of microtubules. Immunofluorescence microscopy using anti-α-tubulin antibody indicated that disassembly of axonemal microtubules of cilia became within 1.5 h after encystment induction, and resorbed in 3 days. Although the cytoplasmic microtubules failed to be visualized clearly, encystmentdependent globulation of cells was promoted by taxol, an inhibitor of disassembly of microtubules. It is possible that a temporary formation of cytoplasmic microtubules may be involved in cell globulation.

The phosphorylation level of actin (43 kDa) became slightly elevated just after encystment induction. Lepidosomes, the sticky small globes surrounding encysting cells, were vividly stained with Acti-stain 555 phalloidin, suggesting that 43-kDa actin or its homologues may be contained in lepidosomes.

Keywords

resting cyst formation, microtubules, actin, lepidosomes, resorption of cilia.

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Information

Information: Acta Protozoologica, 2020, Volume 59, Issue 3-4, pp. 107 - 120

DOI: https://doi.org/10.4467/16890027AP.20.009.13264

Article type: Original article

Authors

Yoichiro Sogame

National Institute of Technology Fukushima College, Iwaki, Fukushima Japan

Katsuhiko Kojima

Department of Microbiology and Immunology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan

Toshikazu Takeshita

Department of Microbiology and Immunology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan

Shiho Kikuchi

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Yuto Shimada

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Rikiya Nakamura

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Mikihiko Arikawa

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Seiji Miyata

Department of Applied Biology, Kyoto Institute of Technology, Kyoto 606-8585, Japan

Eiji Kinoshita

Department of Functional Molecular Science, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Hiroshima 734-8553, Japan

Futoshi Suizu

Oncology Pathology, Department of Pathology and Host-Defense, Faculty of Medicine, Kagawa University, Takamatsu 761-0793, Japan

Tatsuomi Matsuoka

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Published at: 17.03.2021

Received at: 22.04.2020

Accepted at: 05.08.2020

Article status: Open

Licence: CC BY-NC-ND

Percentage share of authors:

Yoichiro Sogame (Author) - 9%

Katsuhiko Kojima (Author) - 9%

Toshikazu Takeshita (Author) - 9%

Shiho Kikuchi (Author) - 9%

Yuto Shimada (Author) - 9%

Rikiya Nakamura (Author) - 9%

Mikihiko Arikawa (Author) - 9%

Seiji Miyata (Author) - 9%

Eiji Kinoshita (Author) - 9%

Futoshi Suizu (Author) - 9%

Tatsuomi Matsuoka (Author) - 10%

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<p> Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of <em>Colpoda cucullus</em> Nag-1 and Cytoskeletal Dynamics</p>

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