Investigation of mechanisms for stress reparation under cryptobiosis in a unicellular organism

• Project/Area Number: 19K16193
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 44050:Animal physiological chemistry, physiology and behavioral biology-related
• Research Institution: Fukushima National College of Technology
• Principal Investigator: Sogame Yoichiro 福島工業高等専門学校, 化学・バイオ工学科, 助教 (60769850)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 無代謝休眠 / 原生生物 / 繊毛虫 / 細胞修復 / ガンマ線 / 休眠シスト / 環境ストレス / 耐性 / ストレス応答 / RNA-Seq / NGS / 単細胞生物 / 放射線 / 環境応答 / 適応進化 / シスト / 細胞ストレス / ストレス修復

Outline of Research at the Start

真核単細胞生物コルポーダは、休眠シストを形成することにより、驚異的な環境ストレス耐性を獲得する。一方、休眠シスト形成過程では、代謝などの生物的特徴は極限まで消失している。申請者は、コルポーダの休眠シスト形成過程において、代謝が不活性化しているにも関わらず、細胞傷害を修復することを明らかにした。本研究では、その休眠過程における細胞ストレス傷害修復機構の分子機構を明らかにすることを目的とする。

Outline of Final Research Achievements

In this study, I analyzed mechanisms for cell repair　of unicellular ciliate Colpoda cucullus in the process of resting cyst formation. I found that Colpoda resting cysts tolerate to more than 8000 Gy gamma radiation and proteins damaged by gamma radiation can be repaired in the cryptobiotic process of the resting cyst formation. In addition, RNA-Seq using next-generation sequencing revealed that some mechanisms controlｌing the resting cyst formation, including the cellular repair mechanism. Many genes whose expression level is up-regulated or down-regulated in resting cysts were identified.

Academic Significance and Societal Importance of the Research Achievements

休眠シストは、古くからその耐性に関して研究されてきた。これまでに、乾燥、高温、低温（凍結）、酸と塩基、UVなど様々な耐性があることが報告されている。本研究では、コルポーダの休眠シストに強いガンマ線耐性があることを明らかにし、休眠シストの耐性に関する知見をさらに広げることができた。また、休眠シストは、ミトコンドリアの膜電位が停止することなどから無代謝休眠の状態であると考えられてきたが、ガンマ線による細胞の障害を修復していることを明らかにした。その修復機構に始まり、休眠シスト形成の現象を司る因子（遺伝子）も同定することができ、休眠シストの理解にさらに繋がったと考えられる。

Research Products

• [Int'l Joint Research] ブランデンブルク工科大学(ドイツ)
• [Journal Article] An elevation transect study of testate amoeba communities up to 4000?m a.s.l. on Mount Kinabalu, Borneo (2022)
  • Author(s): Wanner Manfred、Sogame Yoichiro、Shimizu Miki
  • Journal Title: European Journal of Protistology Volume: 83 Pages: 125868-125868
  • DOI: 10.1016/j.ejop.2022.125868
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Distribution of soil testate amoeba assemblages along an elevation gradient on Mount Fuji (Japan) (2022)
  • Author(s): Andrey N. Tsyganov, Anatoly A. Bobrov, Satoshi D. Shimano, Edward, A.D. Mitchell, Yasuo Hagiwara, Adeline A.J. Wall, Natalia G. Mazei, Viktor A. Chernyshov, Manfred Wanner, Yuantan Zhong, Yoichiro Sogame, Yuri A. Mazei
  • Journal Title: European Journal of Protistology Volume: 84 Pages: 125894-125894
  • DOI: 10.1016/j.ejop.2022.125894
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Testate amoebae from two low-lying tropical islets of Tuvalu, South Pacific (2021)
  • Author(s): Wanner Manfred、Sogame Yoichiro、Shimizu Miki
  • Journal Title: Pedobiologia Volume: 87-88 Pages: 150732-150732
  • DOI: 10.1016/j.pedobi.2021.150732
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] 土壌繊毛虫コルポーダの陸上環境への適応戦略:休眠シスト形成とその環境ストレス耐性 (2021)
  • Author(s): 十亀陽一郎,島野智之,松岡光,有川幹彦,水津太,松岡達臣
  • Journal Title: 原生生物 Volume: 4 Pages: 5-14
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics (2021)
  • Author(s): Sogame Yoichiro、Kojima Katsuhiko、Takeshita Toshikazu、Kikuchi Shiho、Shimada Yuto、Nakamura Rikiya、Arikawa Mikihiko、Miyata Seiji、Kinoshita Eiji、Suizu Futoshi、Matsuoka Tatsuomi
  • Journal Title: Acta Protozoologica Volume: 59 Issue: 3-4 Pages: 107-120
  • DOI: 10.4467/16890027ap.20.009.13264
  • Peer Reviewed / Open Access
• [Journal Article] Isolation of Photobacterium kishitanii Taigaleon from a local fish Mehikari (Greeneye) found near Iwaki City Japan, and possible application for water quality assessment (2020)
  • Author(s): Ryota Saito , Tatsuya Sakai , Taiga Shimizu , Kaito Mizumachi , Ryota Koizumi , Taiki Ono , Yoichiro Sogame
  • Journal Title: Asian Journal of Microbiology, Biotechnology and Environmental Science Volume: 22 Pages: 12-21
  • Peer Reviewed
• [Journal Article] Tolerance of Colpoda cucullus Nag-1 wet resting cysts to extreme pH (pH 1 and 2 13): Implications of less permeability of the cyst membrane to H+ and OH- (2020)
  • Author(s): Rikiya Nakamura , Yoichiro Sogame , Mikihiko Arikawa , Futoshi Suizu , Tatsuomi Matsuoka
  • Journal Title: Journal of Protozoology Research Volume: 30 Pages: 38-46
  • Peer Reviewed / Open Access
• [Journal Article] Spontaneous immobilization of both a fluorescent dye and a functional sugar during the fabrication of submicron-sized PMMA particles in an aqueous solution (2020)
  • Author(s): Yamauchi Noriko、Yatabe Risa、Iino Haruna、Nagatsuka Michi、Sogame Yoichiro、Ogata Makoto、Kobayashi Yoshio
  • Journal Title: Colloids and Surfaces A: Physicochemical and Engineering Aspects Volume: 604 Pages: 125299-125299
  • DOI: 10.1016/j.colsurfa.2020.125299
  • Peer Reviewed
• [Journal Article] Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate Colpoda cucullusNag-1 at －65 ˚C: Possible Involvement of Ultra-Antifreeze Polysaccharides (2020)
  • Author(s): Tatsuomi MATSUOKA , Yoichiro SOGAME , Rikiya NAKAMURA , Yuya HASEGAWA , Mikihiko ARIKAWA , Futoshi SUIZU
  • Journal Title: Acta Protozoologica Volume: 59 Issue: 3-4 Pages: 141-147
  • DOI: 10.4467/16890027ap.20.011.13266
  • Peer Reviewed
• [Journal Article] Comparison of the morphology and viability of gamma irradiated vegetative cells, wet cysts, and dry cysts of the soil ciliate Colpoda cucullus (2020)
  • Author(s): Ryota Saito , Tatsuya Sakai , Ryota Koizumi , Taiga Shimizu , Taiki Ono , Shuntaro Hakozaki , Sena Kobayashi , Yuta Saito , Yoichiro Sogame
  • Journal Title: Journal of Protozoology Research Volume: 30 Pages: 20-30
  • Peer Reviewed
• [Journal Article] Gamma Radiation Tolerance and Protein Carbonylation Caused by Irradiation of Resting Cysts in the Free-living Ciliated Protist Colpoda cucullus (2020)
  • Author(s): Saito Ryota、Koizumi Ryota、Sakai Tatsuya、Shimizu Taiga、Ono Taiki、Sogame Yoichiro
  • Journal Title: Acta Protozoologica Volume: 59 Issue: 2 Pages: 67-75
  • DOI: 10.4467/16890027ap.20.006.12674
  • Peer Reviewed
• [Journal Article] A New Testate Amoeba, Matsakision ogawaraensis sp. nov. (Silicofilosea: Incertae sedis Euglyphida) from Lake Shore Sand of Northern Japan (2020)
  • Author(s): Aoki Yoshiyuki、Sogame Yoichiro、Wanner Manfred、Mazei Yuri、Shimano Satoshi D.
  • Journal Title: Acta Protozoologica Volume: 59 Issue: 2 Pages: 99-105
  • DOI: 10.4467/16890027ap.20.008.12676
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Recovery of proliferative capability in gamma irradiated Colpoda cucullus (ciliated protist) resting cysts and its radiation hormesis (2020)
  • Author(s): Ryota Saito, Ryota Koizumi, Tatsuya Sakai, Taiga Shimizu, Taiki Ono, Manfred Wanner, Toshio Takyu, Toshihiko Tanaka, Yoichiro Sogame
  • Journal Title: Protistology Volume: 14 Issue: 3 Pages: 160-171
  • DOI: 10.21685/1680-0826-2020-14-3-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Lepidosomes acquire fluorescence after encystation: Including additional notes of morphological events during encystation and reconsideration of the morphological features in the ciliate <i>Colpoda cucullus</i> (2019)
  • Author(s): Yoichiro Sogame, Ryota Saito, Tatsuya Sakai, Taiga Shimizu, Taiki Ono, Ryota Koizumi, Kaito Mizumachi
  • Journal Title: The Journal of Protozoology Research Volume: 29 Issue: 1-2 Pages: 51-62
  • DOI: 10.32268/jprotozoolres.29.1-2_51
  • NAID: 130007813482
  • ISSN: 0917-4427, 2434-7027
  • Peer Reviewed / Open Access
• [Journal Article] Evidence of Stress Recovery in Free-Living Ciliate Colpoda cucullus: The Repair Capability of Resting Cysts to Damage Caused by Gamma Irradiation (2019)
  • Author(s): Yoichiro Sogame, Ryota Saito, Ryota Koizumi, Taiga Shimizu, Taiki Ono
  • Journal Title: Acta Protozoologica Volume: 58 Issue: 1 Pages: 25-29
  • DOI: 10.4467/16890027ap.19.006.10837
  • Peer Reviewed / Open Access
• [Presentation] 繊毛虫Colpoda休眠シストにおけるATPの定量解析 (2022)
  • Author(s): 矢吹一真, 齋藤剛瑠, 山野邊裕樹, 齊藤瞭汰, 清水大雅, 箱﨑惇太朗, 十亀陽一郎
  • Organizer: 第55回日本原生生物学会大会 2022年9月
• [Presentation] 繊毛虫Colpodaの休眠シスト形成と環境ストレス耐性 (2022)
  • Author(s): 十亀陽一郎
  • Organizer: 第55回日本原生生物学会大会 2022年9月
• [Presentation] 真核単細胞生物の休眠シストにおける無代謝休眠の分子機構解明 (2022)
  • Author(s): 十亀陽一郎
  • Organizer: 日本比較生理生化学会第44回 2022年11月
• [Presentation] 単細胞生物繊毛虫 Colpoda 休 眠シスト形成過程における代謝活性の解析 (2022)
  • Author(s): 十亀陽一郎, 山野邊裕樹, 齊藤瞭汰, 清水大雅, 矢吹一真, 齋藤剛瑠, 箱﨑惇太朗
  • Organizer: 第93回日本動物学会 2022年9月8日
• [Presentation] Alteration of gene expression contributing to high salinity tolerance of resting cysts in Colpoda cucullus (2021)
  • Author(s): Yoichiro SOGAME , Ryota SAITO , Ryota KOIZUMI , Tomohiro SUZUKI , Tatsuya SAKAI , Hiroki , YAMANOBE , Taiga SHIMIZU , Akiko ONO , Tetsuya Aoyama , Toshihiko TANAKA , Manfred WANNER , Kozo WATANABE
  • Organizer: 日本比較生理生化学会第43回
• [Presentation] Colpoda cucullus休眠シスト の塩耐性：シストは海を渡るだろうか？ (2021)
  • Author(s): 十亀陽一郎 , 齊藤 瞭汰 , 小泉 亮太 , 鈴木 智大 , 酒井 達弥 , 山野邊 裕樹 , 清水 大雅 , 小野 晶子 , 田中 利彦 , 青山 哲也 , ワーナー マンフレッド , 渡辺 幸三
  • Organizer: 第54回日本原生生物学会大会
• [Presentation] Tolerance of Colpoda cucullus Nag-1 wet resting cysts to extreme pH (pH 1 and 13): Implications of less permeability of the cyst membrane to H+ and OH- (2021)
  • Author(s): Rikiya Nakamura , Yoichiro Sogame , Mikihiko Arikawa , Futoshi Suizu , Tatsuomi Matsuoka
  • Organizer: The 4th Asian Congress of Protistology（ACOP-IV)
  • Int'l Joint Research
• [Presentation] High Salinity Tolerance and Alteration of Gene Expression during Resting Cyst formation of Freshwater Soil Ciliate Colpoda cucullus (2021)
  • Author(s): Yoichiro SOGAME , Ryota SAITO , Ryota KOIZUMI , Tomohiro SUZUKI , Tatsuya SAKAI , Hiroki YAMANOBE , Akiko ONO , Sena KOBAYASHI , Shuntaro HAKOZAKI , Yuta SAITO , Toshihiko TANAKA , Kozo WATANABE
  • Organizer: Water and Environment Technology Conference Online2021
  • Int'l Joint Research
• [Presentation] Analysis of Influence of Gamma Irradiation on Cell Viability, Proliferative Capability, Morphology, and Proteins in Resting Cysts of Colpoda (2020)
  • Author(s): Ryota Saito , Ryota Koizumi , Tatsuya Sakai , Taiga Shimizu , Taiki Ono, Yoichiro Sogame
  • Organizer: Water and Environment Technology Conference Online 2020
  • Int'l Joint Research
• [Presentation] 十亀陽一郎、小泉亮太、齊藤瞭汰、酒井達弥、清水大雅、中村瑠奈、鈴木智大 (2020)
  • Author(s): 土壌繊毛虫Colpoda休眠シストにおける細胞傷害修復と発現遺伝子の解析
  • Organizer: 第54回日本水環境学会年会 2020年3月
• [Presentation] 土壌繊毛虫Colpoda cucullusシスト形成におけるシスト特異的細胞構造形成過程の解析 (2020)
  • Author(s): 小泉亮太、酒井達弥、齊藤暸汰、清水大雅、小野泰輝、十亀陽一郎
  • Organizer: 第54回日本水環境学会年会 2020年3月
• [Presentation] 繊毛虫Colpoda休眠シストにおけるガンマ線耐性と細胞修復 (2019)
  • Author(s): 十亀陽一郎, 齊藤瞭汰, 小泉亮太, 清水大雅, 小野泰輝, 酒井達弥
  • Organizer: 第52回日本原生生物学会
• [Presentation] A New Insight into Cryptobiosis- Cell Repair in Ametabolism (2019)
  • Author(s): Yoichiroo Sogame, Ryota Saito, Ryota Koizumi
  • Organizer: International Caparica Conference on Analytical Proteomics
  • Int'l Joint Research
• [Presentation] Tolerance to Ionizing radiation and Irradiation Damage Repair in the Unicellular Eukaryote Colpoda cucullus (2019)
  • Author(s): Yoichiro SOGAME, Ryota SAITO, Ryota KOIZUMI, Taiga SHIMIZU, Tatsuya SAKAI, Taiki ONO, Toshio TAKYU
  • Organizer: Water and Environment Technology Conference
  • Int'l Joint Research
• [Presentation] Morphological Events in the Formation of Cyst-specific Cell Structures and Acquisition of Autofluorescence during the Encystment Process in Colpoda cucullus (2019)
  • Author(s): Tatsuya SAKAI, Taiga SHIMIZU, Ryota KOIZUMI, Ryota SAITO, Taiki ONO, Yoichiro SOGAME
  • Organizer: Water and Environment Technology Conference
  • Int'l Joint Research
• [Presentation] Extreme Gamma Radiation Tolerance of Colpoda cucullus: Physiological and Morphological Analysis of Irradiated Cysts (2019)
  • Author(s): Ryota SAITO, Ryota KOIZUMI, Taiga SHIMIZU, Taiki ONO, Tatsuya SAKAI, Toshio TAKYU, Yoichiro SOGAME
  • Organizer: Water and Environment Technology Conference
  • Int'l Joint Research
• [Presentation] Effects of Cell Repair on the Excystment of Irradiated Cysts and Cell Growth of Vegetative Cells Excysted from Irradiated Cysts in Colpoda cucullus (2019)
  • Author(s): Ryota Koizumi, Ryota Saito, Taiga Shimizu, Toshio Takyu, Yoichiro Sogame
  • Organizer: Water and Environment Technology Conference
  • Int'l Joint Research